OBJECTIVE:To clarify the effect of Hamayou(Oviductus Ranae) protein hydrolysate(ORPH) on depression and its exact underlying mechanism from a new perspective. METHODS:We used the Chronic Unpredictable Mild Stress(CUMS...OBJECTIVE:To clarify the effect of Hamayou(Oviductus Ranae) protein hydrolysate(ORPH) on depression and its exact underlying mechanism from a new perspective. METHODS:We used the Chronic Unpredictable Mild Stress(CUMS) method to prepare a mouse model of depression and lipopolysaccharide(LPS) to prepare a model of BV2 cellular inflammation to investigate the antidepressant effect and mechanism of action of ORPH. Behavioral changes in mice and cerebral blood flow were detected by behavioral experiments and scatter imaging. Levels of corticosterone(CORT), proinflammatory cytokines and neurotransmitter were detected by enzyme-linked immunosorbent assay. Furthermore, hematoxylin-eosin staining, Tunel staining were used to evaluate the effect of ORPH. The distribution and expression of ionized calcium bindingadaptor molecule-1(Iba-1) in mouse hippocampal tissue and BV2 cells were detected by immunofluorescence. Mitogen-activated protein kinase(MAPK) pathway related protein expression was detected by Western blot. RESULTS:ORPH improved depression-like behavior, ameliorated brain tissue damage and apoptosis, and inhibited microglia activation in brain tissue in mice. In addition, ORPH reduced expression of B-cell lymphoma-2(Bcl-2)-associated X(Bax), cysteinyl aspartate specific proteinase 3(Caspase3), cysteinyl aspartate specific proteinase 9(Caspase9), nuclear factor-kappa B(NF-κB), phosphorylation-p38(p-p38), phosphorylation-Jun Nterminal kinase(p-JNK) proteins, and increased expression of Bcl-2, inhibitory kappa B alpha(IκB-α), phosphorylation-extracellular regulated protein kinases 1/2(p-ERK1/2) proteins. On the other hand, there were fewer Iba-1-positive cells, lower expression of NF-κB, pp38, p-JNK and p-ERK1/2 proteins, and higher expression of IκB-α proteins in BV2 cells in the ORPH group. In addition, ORPH increased 5-hydroxytryptamine, norepinephrine levels and decreased CORT, interleukin-1β(IL-1β), interleukin-6(IL-6), tumor necrosis factor-α(TNF-α) levels. CONCLUSION:ORPH was able to improve depressionlike behaviors and that it took effects by promoting cerebral blood flow, inhibition of hypothalamic-pituitaryadrenal axis overactivation, improving the structural damage of hippocampal tissues, and inhibiting the inflammatory response. ORPH can reduced neuronal damage and inhibiting apoptosis by promoting the MAPK pathway.展开更多
OBJECTIVE:To confirm the therapeutic effect and mechanism of Jingui Shenqi pill(金匮肾气丸,JGSQP)on cardiorenal syndrome.METHODS:Doxorubicin was used to build heart-kidney coinjury rat model.After the modeling was com...OBJECTIVE:To confirm the therapeutic effect and mechanism of Jingui Shenqi pill(金匮肾气丸,JGSQP)on cardiorenal syndrome.METHODS:Doxorubicin was used to build heart-kidney coinjury rat model.After the modeling was completed,JGSQP gavage intervention was performed.The cardiac function of rats in each group was evaluated by ultrasound detection.Serum of rats was collected and examined for markers of heart and kidney damage.Enzyme linked immunosorbent assay detected serum inflammatory factors interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)expression.Quantitative real-time polymerase chain reaction(PCR)and Western blot detected the changes of related genes and proteins.RESULTS:JGSQP significantly increased left ventricular ejection fraction(EF)and left ventricular shortening fraction(FS)values,decreased the heart and kidney damage markers and fibrosis levels(P<0.05).Furthermore,it can reduce IL-1β,IL-6,and TNF-αinflammatory expression(P<0.05).Mechanistically,JGSQP significantly inhibited the expression of key genes and proteins of mitogen-activated protein kinase(MAPK)signaling pathway(P<0.05).CONCLUSIONS:Jingui Shenqi pill can exert therapeutic effects on cardiorenal syndrome by inhibiting the activation of the MAPK signaling pathway and inflammatory responses.展开更多
Objective To evaluate the antagonistic effects of N-acetylcysteine(NAC)on mitogen-activated protein kinases(MAPK)pathway activation,oxidative stress and inflammatory responses in rats with lung injury induced by fine ...Objective To evaluate the antagonistic effects of N-acetylcysteine(NAC)on mitogen-activated protein kinases(MAPK)pathway activation,oxidative stress and inflammatory responses in rats with lung injury induced by fine particulate matter(PM2.5).Methods Forty eight male Wistar rats were randomly divided into six groups:blank control group(C1),water drip control group(C2),PM2.5 exposed group(P),low-dose NAC treated and PM2.5 exposed group(L),middle-dose NAC treated and PM2.5 exposed group(M),and high-dose NAC treated and PM2.5 exposed group(H).PM2.5 suspension(7.5 mg/kg)was administered tracheally once a week for four times.NAC of 125 mg/kg,250 mg/kg and 500 mg/kg was delivered intragastrically to L,M and H group respectively by gavage(10 ml/kg)for six days before PM2.5 exposure.The histopathological changes and human mucin 5 subtype AC(MUC5AC)content in lung tissue of rats were evaluated.We investigated IL-6 in serum and bronchoalveolar lavage fluid(BALF)by Enzyme-linked immunosorbent assay(ELISA),MUC5AC in lung tissue homogenate by ELISA,glutathione peroxidase(GSH-PX)in serum and BALF by spectrophotometry,and the expression of p-ERK1/2,p-JNK1/2 and p-p38 proteins by Western blot.All the measurements were analyzed and compared statistically.Results Lung tissue of rats exposed to PM2.5 showed histological destruction and increased mucus secretion of bronchial epithelial cells.Rats receiving NAC treatment showed less histological destruction and mucus secretion.Of P,L,M and H group,MUC5AC in lung tissue,IL-6 in serum and BALF were higher than controls(C1 and C2)(all P<0.05),with the highest levels found in the P group and a decreasing trend with increase of NAC dose.The activity of GSH-PX in serum and BALF of PM2.5 exposed rats(P,L,M and H)was lower than that of controls(all P<0.05),with higher activities found in NAC treated rats(L,M,and H),and an increasing trend with increase of NAC dose.The expressions of p-ERK1/2,p-JNK1/2 and p-p38 proteins in PM2.5 exposed lung tissue(P,L,M and H)was higher than controls(all P<0.05),with decreased levels and dose dependent downregulation found in NAC treated rats.Conclusion NAC can antagonize major MAPK pathway activation,lung oxidative stress and inflammatory injury induced by PM2.5 in rats.展开更多
Marsdenia tenacissima extract(MTE, trade name: Xiao-Ai-Ping injection) is an extract of a single Chinese plant medicine. It has been used for the treatment of cancer in China for decades, especially for esophageal can...Marsdenia tenacissima extract(MTE, trade name: Xiao-Ai-Ping injection) is an extract of a single Chinese plant medicine. It has been used for the treatment of cancer in China for decades, especially for esophageal cancer and other cancers in the digestive tract. In the present study, the potential mechanism for MTE's activity in esophageal cancer was explored. The effects of MTE on the proliferation of human esophageal cancer cells(KYSE150 and Eca-109) were investigated by the MTT assay, the Brd U(bromodeoxyuridine) incorporation immunofluorescence assay, and flow cytometric analysis. MTE inhibited cell proliferation through inducing G0/G1 cell cycle arrest in KYSE150 and Eca-109. Western blot analysis was employed to determine protein levels in the MTE treated cells. Compared with the control cells, the expression levels of the cell cycle regulatory proteins cyclin D1/D2/D3, cyclin E1, CDK2/4/6(CDK: cyclin dependent kinase), and p-Rb were decreased significantly in the cells treated with MTE at 40 mg·m L-1. In addition, MTE had an inhibitory effect on the MAPK(mitogen-activated protein kinase) signal transduction pathway, including ERK(extracellular signal-regulated kinase), JNK(c-Jun N-terminal kinase), and p38 MAPK. Moreover, MTE showed little additional effects on the regulation of cyclin D1/D3, CDK4/6, and p-Rb when the ERK pathway was already inhibited by the specific ERK inhibitor U0126. In conclusion, these data suggest that MTE inhibits human esophageal cancer cell proliferation through regulation of cell cycle regulatory proteins and the MAPK signaling pathways, which is probably mediated by the inhibition of ERK activation.展开更多
Electroacupuncture attenuates cerebral hypoxia and neuronal apoptosis induced by cerebral ischemia/reperfusion injury.To further identify the involved mechanisms,we assumed that electroacupuncture used to treat cerebr...Electroacupuncture attenuates cerebral hypoxia and neuronal apoptosis induced by cerebral ischemia/reperfusion injury.To further identify the involved mechanisms,we assumed that electroacupuncture used to treat cerebral ischemia/reperfusion injury was associated with the p38 mitogen-activated protein kinase（MAPK） signaling pathway.We established rat models of cerebral ischemia/reperfusion injury using the modified Zea-Longa＇s method.At 30 minutes before model establishment,p38 MAPK blocker SB20358 was injected into the left lateral ventricles.At 1.5 hours after model establishment,electroacupuncture was administered at acupoints of Chize（LU5）,Hegu（LI4）,Zusanli（ST36）,and Sanyinjiao（SP6） for 20 minutes in the affected side.Results showed that the combination of EA and SB20358 injection significantly decreased neurologic impairment scores,but no significant differences were determined among different interventional groups.Hematoxylin-eosin staining also showed reduced brain tissue injuries.Compared with the SB20358 group,the cells were regularly arranged,the structures were complete,and the number of viable neurons was higher in the SB20358 ＋ electroacupuncture group.Terminal deoxynucleotidyl transferase（Td T）-mediated d UTP nick-end labeling assay showed a decreased apoptotic index in each group,with a significant decrease in the SB20358 ＋ electroacupuncture group.Immunohistochemistry revealed reduced phosphorylated p38 expression at 3 days in the electroacupuncture group and SB20358 ＋ electroacupuncture group compared with the ischemia/reperfusion group.There was no significant difference in phosphorylated p38 expression between the ischemia/reperfusion group and SB20358 group.These findings confirmed that the electroacupuncture effects on mitigating cerebral ischemia/reperfusion injury are possibly associated with the p38 MAPK signaling pathway.A time period of 3 days could promote the repair of ischemic cerebral nerves.展开更多
AIM: To investigate the expression of mitogen-activated protein kinases (MAPKs) and its upstream protein kinase in human gastric cancer and to evaluate the relationship between protein levels and clinicopathological p...AIM: To investigate the expression of mitogen-activated protein kinases (MAPKs) and its upstream protein kinase in human gastric cancer and to evaluate the relationship between protein levels and clinicopathological parameters. METHODS: Western blot was used to measure the expression of extracellular signal-regulated kinase (ERK)-1, ERK-2, ERK-3, p38 and mitogen or ERK activated protein kinaseMEK-1 proteins in surgically resected gastric carcinoma, adjacent normal mucosa and metastatic lymph nodes from 42 patients. Immunohistochemistry was employed for their localization. RESULTS: Compared with normal tissues, the protein levels of ERK-1 (integral optical density value 159 526?5 760 vs 122 807±65 515, P= 0.001), ERK-2 (168 471±95 051 vs 120 469±72 874, P<0.001), ERK-3 (118 651±71 513 vs 70 934±68 058,P<0.001), P38 (104 776±51 650 vs 82 930±40 392, P= 0.048) and MEK-1 (116 486±45 725 vs 101 434±49 387, P = 0.027) were increased in gastric cancer tissues. Overexpression of ERK-3 was correlated to TNM staging [average ratio of integral optic density (IOD)tumor: IODnormal in TNM I, II, III, IV tumors was 1.43±0.34, 5.08±3.74, 4.99±1.08, 1.44±1.02, n = 42, P= 0.023] and serosa invasion (4.31±4.34 vs 2.00±2.03, P = 0.037). In poorly differentiated cancers (n = 33), the protein levels of ERK-1 and ERK-2 in stage III and IV tumors were higher than those in stage I and II tumors (2.64+3.01 vs 1.01±0.33, P= 0.022; 2.05±1.54 vs1.24±0.40, P= 0.030). Gastric cancer tissues with either lymph node involvement (2.49±2.91 vs1.03±0.36, P= 0.023; 1.98±1.49vs1.24±0.44, P= 0.036) or serosa invasion (2.39±2.82 vs 1.01±0.35, P= 0.022; 1.95±1.44 vs1.14±0.36, P=0.015) expressed higher protein levels of ERK-1 and ERK-2. In Borrmann II tumors, expression of ERK-2 and ERK-3 was increased compared with Borrmann III tumors (2.57±1.86 vs1.23±0.60, P= 0.022; 5.50±5.05 vs1.83±1.21, P= 0.014). Borrmann IV tumors expressed higher p38 protein levels. No statistically significant difference in expression of MAPKs was found when stratified to tumor size or histological grade (P>0.05). Protein levels of ERK-2, ERK-3 and MEK-1 in metastatic lymph nodes were 2-7 folds higher than those in adjacent normal mucosa. The immunohistochemistry demonstrated that ERK-1, ERK-2, ERK-3, p38 and MEK-1 proteins were mainly localized in cytoplasm. The expression of MEK-1 in gastric cancer cells metastasized to lymph nodes was higher than that of the primary site. CONCLUSION: MAPKs, particularly ERK subclass are overexpressed in the majority of gastric cancers. Overexpression of ERKs is correlated to TNM staging, serosa invasion, and lymph node involvement. The overexpression of p38 most likely plays a prominent role in certain morphological subtypes of gastric cancers. MEK-1 is also overexpressed in gastric cancer, particularly in metastatic lymph nodes. Upregulation of MARK signal transduction pathways may play an important role in tumorigenesis and metastatic potential of gastric cancer.展开更多
Cold-inducible RNA-binding protein （CIRP） is an RNA-binding protein that is expressed in normal testes and downregulated after heat stress caused by cryptorchidism, varicocele or environmental temperatures. The purp...Cold-inducible RNA-binding protein （CIRP） is an RNA-binding protein that is expressed in normal testes and downregulated after heat stress caused by cryptorchidism, varicocele or environmental temperatures. The purpose of this study was to investigate the functions of CIRP in the testes. We employed RNAi technique to knock down the expression of CIRP in the testes, and performed haematoxylin and eosin staining to evaluate morphological changes following knockdown. Germ cell apoptosis was examined by terminal deoxynucleotidal transferase-mediated dUTP nick end labelling （TUNEL） assay, and mitogen-activated protein kinase （MAPK） signalling pathways were investigated by Western blotting to determine the possible mechanism of apoptosis. We found that using siRNA is a feasible and reliable method for knocking down gene expression in the testes. Compared to controls, the mean seminiferous tubule diameter （MSTD） and the thickness of the germ cell layers decreased following siRNA treatment, whereas the percentage of apoptotic seminiferous tubules increased. The p44/p42, p38 and SAPK/JNK MAPK pathways were activated after downregulation of CIRP. In conclusion, we discovered that downregulation of CIRP resulted in increased germ cell apoptosis, possibly viathe activation of the p44/p42, p38 and SAPK/JNK MAPK pathways.展开更多
OBJECTIVE: To explore the effects of evodiamine on ovarian cancer cells and the mechanisms underlying such effects.METHODS: Human ovarian cancer cells HO-8910 PM were treated with evodiamine at 0, 1.25,2.5, and 5 μM ...OBJECTIVE: To explore the effects of evodiamine on ovarian cancer cells and the mechanisms underlying such effects.METHODS: Human ovarian cancer cells HO-8910 PM were treated with evodiamine at 0, 1.25,2.5, and 5 μM for 1-4 d. 3-(4,5-Dimethiylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay was used to detect the growth inhibition rate of evodiamine-treated HO-8910 PM cells. The cell cycle was observed via propidium iodide(PI) staining. Apoptosis induction was assessed via Annexin V-fluorescein isothiocyanate/propidium iodide(Annexin V-FITC/PI) double staining assay. To verify the mechanism of apoptosis, caspase-dependent apoptotic pathway-related protein was detected by Western blot analysis. The expression levels of mitogen-activated protein kinase(MAPK)and/or phosphatidylinositol-3-kinase(PI3K)/pro-tein kinase B(Akt) pathway-related proteins were also investigated.RESULTS: Evodiamine significantly inhibited the proliferation of HO-8910 PM cells in a dose- and time-dependent manner. Evodiamine induced G2/M arrest with an increase of cyclin B1 level, and promoted cell apoptosis with a decrease of B cell lymphoma/lewkmia-2(Bcl-2) and an increase of Bcl-2-associated X protein(Bax) level. In addition,evodiamine treatment led to the activation of caspase-8, caspase-9, and caspase-3 and the cleavage of poly(ADP-ribose)-polymerase(PARP). Evodiamine targeted the MAPK and/or PI3K/Akt pathways by reducing the expression and activity of PI3 K, Akt, and extracellular signal-regulated kinase mitogen-activated protein kinase(ERK1/2 MAPK)and the activity of p38 MAPK.CONCLUSION: Evodiamine can inhibit the growth of ovarian cancer cells by G2/M arrest and intrinsic and extrinsic apoptosis. In addition, evodiamine-induced PI3K/Akt, ERK1/2 MAPK, and p38 MAPK signaling may be involved in cell death.展开更多
AIM: To investigate the role of p38 mitogen-activated protein kinase in rat small intestine after ischemia-reperfusion (I/R)insult and the relationship between activation of p38 MAPK and apoptotic cell death of intest...AIM: To investigate the role of p38 mitogen-activated protein kinase in rat small intestine after ischemia-reperfusion (I/R)insult and the relationship between activation of p38 MAPK and apoptotic cell death of intestine.METHODS: Ninety Wistar rats were divided randomly into three groups, namely sham-operated group (C), I/R vehicle group (R) and SB203580 pre-treated group(S).In groups R and S, the superior mesenteric artery(SMA)was separated and occluded for 45 min, then released for reperfusion for0.25, 0.5, 1, 2, 6, 12 and 24 h. In group C, SMA was separated without occlusion. Plasma D-lactate levels were examined and histological changes were observed under a light microscope. The activity of p38 MAPK was determined by Western immunoblotting and apoptotic cells were detected by the terminal deoxynucleotidyl transferase (TdT)-mediated dUDP-biotin nick end labeling (TUNEL).RESULTS: Intestinal ischemia followed by reperfusion activated p38 MAPK, and the maximal level of activation (7.3-fold vs sham-operated group) was reached 30 min after I/R. Treatment with SB 203580, a p38 MAPK inhibitor,reduced intestinal apoptosis (26.72±3.39% vs62.50±3.08%in I/R vehicle, P＜0.01) and decreased plasma D-lactate level (0.78±0.15 mmol/L in I/R vehicle vs0.42±0.17 mmol/L in SB-treated group) and improved post-ischemic intestinal histological damage.CONCLUSION: p38 MAPK plays a crucial role in the signal transduction pathway mediating post-ischemic intestinal apoptosis, and inhibition of p38 MAPK may attenuate ischemia-reperfusion injury.展开更多
The mitogen-activated protein kinase（MAPK） signaling pathway plays an important role in the regulation of cell growth, proliferation, differentiation, transformation and death. Mitogen-activated protein kinase phosp...The mitogen-activated protein kinase（MAPK） signaling pathway plays an important role in the regulation of cell growth, proliferation, differentiation, transformation and death. Mitogen-activated protein kinase phosphatase 1（MKP1） has an inhibitory effect on the p38 MAPK and JNK pathways, but it is unknown whether it plays a role in Aβ-induced oxidative stress and neuronal inflammation. In this study, PC12 cells were infected with MKP1 sh RNA, MKP1 lentivirus or control lentivirus for 12 hours, and then treated with 0.1, 1, 10 or 100 μM amyloid beta 42（Aβ42）. The cell survival rate was measured using the cell counting kit-8 assay. MKP1, tumor necrosis factor-alpha（TNF-α） and interleukin-1β（IL-1β） m RNA expression levels were analyzed using quantitative real time-polymerase chain reaction. MKP1 and phospho-c-Jun N-terminal kinase（JNK） expression levels were assessed using western blot assay. Reactive oxygen species（ROS） levels were detected using 2′,7′-dichlorofluorescein diacetate. Mitochondrial membrane potential was measured using flow cytometry. Superoxide dismutase activity and malondialdehyde levels were evaluated using the colorimetric method. Lactate dehydrogenase activity was measured using a microplate reader. Caspase-3 expression levels were assessed by enzyme-linked immunosorbent assay. Apoptosis was evaluated using the terminal deoxynucleotidyl transferase d UTP nick end labeling method. MKP1 overexpression inhibited Aβ-induced JNK phosphorylation and the increase in ROS levels. It also suppressed the Aβ-induced increase in TNF-α and IL-1β levels as well as apoptosis in PC12 cells. In contrast, MKP1 knockdown by RNA interference aggravated Aβ-induced oxidative stress, inflammation and cell damage in PC12 cells. Furthermore, the JNK-specific inhibitor SP600125 abolished this effect of MKP1 knockdown on Aβ-induced neurotoxicity. Collectively, these results show that MKP1 mitigates Aβ-induced apoptosis, oxidative stress and neuroinflammation by inhibiting the JNK signaling pathway, thereby playing a neuroprotective role.展开更多
OBJECTIVE:To examine the role and decipher the mechanism of Pingchuan formula(平喘方,PCF)in treating allergic asthma.METHODS:The mice were treated with saline,dexamethasone(DXM)and PCF for 1 week after the asthma mode...OBJECTIVE:To examine the role and decipher the mechanism of Pingchuan formula(平喘方,PCF)in treating allergic asthma.METHODS:The mice were treated with saline,dexamethasone(DXM)and PCF for 1 week after the asthma model was established and their respiratory function including respiratory resistance(RI),pulmonary dynamic compliance(Cdyn)and maximum voluntary ventilation(MVV)were measured.In addition,cellular changes in bronchoalveolar lavage fluid(BALF)and pathological changes in lung biopsy as well as the expression level ofα-smooth muscle actin(α-SMA),transforming growth factor-beta1(TGF-β1)in BALF and interleukin-5(IL-5),interleukin-13(IL-13),tumor necrosis factor-α(TNF-α),interferon-γ(IFN-γ),nuclear factor-kappa B-p65(NF-κBp65),inhibitor-αof nuclear transcription factorκB(IκBα),p38 mitogen-activated protein kinase(p38 MAPK),c-jun n-terminal kinase(JNK)and its phosphorylated proteins in lung tissue were also examined and compared among different groups.RESULTS:Our data suggested that the respiratory functions were significantly improved and the pathological changes ameliorated in the DXM group and the PCF group compared to the model group.Both DXM and PCF effectively decreased the number of eosinophils,lymphocytes,and neutrophils in BAL as well as the secretion ofα-SMA and TGF-β1,IL-5,IL-13,while increased the expression of TNF-αand IFN-γ.Furthermore,our study indicated that the NF-κBp65,IκBα,p38 MAPK and JNK pathways were inhibited under the treatment of PCF.CONCLUSION:Our data indicated that PCF can attenuate the inflammatory response in asthma through inhibiting the NF-κB/MAPK signaling pathway.This study not only supported the use of PCF in allergic asthma in clinic but also shed light upon afurther understanding of the disease pathogenesis.展开更多
AIM: To explore the mechanism by which H pylori causes activation of gastric epithelial cells. METHODS: A VacA (+) and CagA (+) standard H pylori line NCTC 11637 and a human gastric adenocarcinoma derived gastric epit...AIM: To explore the mechanism by which H pylori causes activation of gastric epithelial cells. METHODS: A VacA (+) and CagA (+) standard H pylori line NCTC 11637 and a human gastric adenocarcinoma derived gastric epithelial cell line BGC-823 were applied in the study. MTT assay and 3H-TdR incorporation test were used to detect the proliferation of BGC-823 cells and Western blotting was used to detect the activity and existence of related proteins. RESULTS: Incubation with H pylori extract increased the proliferation of gastric epithelial cells, reflected by both live cell number and DNA synthesis rate. The activity of extracellular signal-regulated protein kinase (ERK) signal transduction cascade increased within 20 min after in- cubation with H pylori extract and appeared to be a sus- tained event. MAPK/ERK kinase (MEK) inhibitor PD98059 abolished the action of H pylori extract on both ERK activity and cell proliferation. Incubation with H pylori extract increased c-Fos expression and SRE-dependent gene expression. H pylori extract caused phosphorylation of several proteins including a protein with molecular size of 97.4 kDa and tyrosine kinase inhibitor genistein inhibited the activation of ERK and the proliferation of cells caused by H pylori extract. CONCLUSION: Biologically active elements in H pylori extract cause proliferation of gastric epithelial cells through activating tyrosine kinase and ERK signal trans- duction cascade.展开更多
BACKGROUND:Paraquat(PQ)-induced acute lung injury(ALI)and pulmonary fi brosis are common diseases with high mortality but without eff ective antidotes in emergency medicine.Our previous study has proved that arctigeni...BACKGROUND:Paraquat(PQ)-induced acute lung injury(ALI)and pulmonary fi brosis are common diseases with high mortality but without eff ective antidotes in emergency medicine.Our previous study has proved that arctigenin suppressed pulmonary fibrosis induced by PQ.We wondered whether arctigenin could also have a protective eff ect on PQ-induced ALI.METHODS:A PQ-induced A549 cell injury model was used,and the effect of arctigenin was determined by a cell counting kit-8(CCK-8)cell viability assay.In addition,terminal deoxynucleotidyl transferase(TdT)-mediated dUTP nick-end labelling(TUNEL)staining assays and mitochondrial membrane potential assays were performed to evaluate the level of cell apoptosis.The generation of reactive oxygen species(ROS)was refl ected by dihydroethidium(DHE)staining and a 2’,7’-dichlorodihy drofluorescein diacetate(DCFH-DA)assay.Moreover,immunoblotting studies were used to assess the expression of mitogen-activated protein kinases(MAPKs)and p38 MAPK.RESULTS:Arctigenin attenuated PQ-induced inhibition of A549 cell viability in a dose-dependent manner.Arctigenin also significantly reduced PQ-induced A549 cell apoptosis,as refl ected by the TUNEL assay and mitochondrial membrane potential assay,which may result from suppressed ROS/p38 MAPK signaling because we found that arctigenin dramatically suppressed ROS generation and p38 MAPK phosphorylation.CONCLUSION:Arctigenin could attenuate PQ-induced lung epithelial A549 cell injury in vitro by suppressing ROS/p38 MAPK-mediated cell apoptosis,and arctigenin might be considered a potential candidate drug for PQ-induced ALI.展开更多
OBJECTIVE: To study the molecular mechanism of Buyang Huanwu Tang( 补阳还五汤)(BHT) protecting retinal ganglion cells(RGCs) from oxygen induced oxidative stress and apoptosis after anterior ischemia. METHODS: In this ...OBJECTIVE: To study the molecular mechanism of Buyang Huanwu Tang( 补阳还五汤)(BHT) protecting retinal ganglion cells(RGCs) from oxygen induced oxidative stress and apoptosis after anterior ischemia. METHODS: In this study, the Chinese herbs of BHT were extracted by first boiling in water, then were filtered, concentrated, and freeze-dried. The chemical profile of BHT extract was determined by liquid chromatography mass spectrometry(LC-MS). H2O2-induced RGC-5 cells were used as a cell model to investigate the protective effect and mechanism of BHT on RGCs. RESULTS: The survival rate of damaged RGC-5 by BHT was significantly increased by the 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazolium-romid method. Fluorescence activating cell sorter(FACS analysis) showed that BHT could significantly reduce apoptosis induced by oxidative stress via the reactive oxygen species(ROS)-mitogen-activated protein kinase(MAPK)-Caspase-3 signal pathway. CONCLUSION: BHT possesses a high antioxidant capacity and could significantly reduce ROS levels of RGC-5 cells damaged by H2O2.Therefore, the present study has provided possible alternative strategies for the prevention and treatment of ischemic optic disease by using traditional Chinese herbal formulas.展开更多
The mortality rate of gastric cancer worldwide is as high as 70%, despite the development of novel therapeutic strategies. One reason for the high mortality is the rapid and uninhibited spread of the disease, such tha...The mortality rate of gastric cancer worldwide is as high as 70%, despite the development of novel therapeutic strategies. One reason for the high mortality is the rapid and uninhibited spread of the disease, such that the majority of patients are diagnosed at a stage when efficient therapeutic treatment is not available. Therefore, in-depth research is needed to investigate the mechanism of gastric cancer metastasis and invasion to improve outcomes and provide biomarkers for early diagnosis. The mitogen-activated protein kinase(MAPK) signaling pathway is widely expressed in multicellular organisms, with critical roles in multiple biological processes, such as cell proliferation, death, differentiation, migration, and invasion. The MAPK pathway typically responds to extracellular stimulation. However, the MAPK pathway is often involved in the occurrence and progression of cancer when abnormally regulated. Many studies have researched the relationship between the MAPK signaling pathway and cancer metastasis and invasion, but little is known about the important roles that the MAPK signaling pathway plays in gastric cancer. Based on an analysis of published data, this review aims to summarize the important role that the MAP kinases play in the invasion and metastasis of gastric cancer and attempts to provide potential directions for further research and clinical treatment.展开更多
Acute pancreatitis(AP) is an inflammatory disease characterized by acute inflammation and necrosis of the pancreatic parenchyma. AP is often associated with organ failure, sepsis, and high mortality. The pathogenesis ...Acute pancreatitis(AP) is an inflammatory disease characterized by acute inflammation and necrosis of the pancreatic parenchyma. AP is often associated with organ failure, sepsis, and high mortality. The pathogenesis of AP is still not well understood. In recent years several papers have highlighted the cellular and molecular events of acute pancreatitis. Pancreatitis is initiated by activation of digestive enzymes within the acinar cells that are involved in autodigestion of the gland, followed by a massive infiltration of neutrophils and macrophages and release of inflammatory mediators, responsible for the local and systemic inflammatory response. The hallmark of AP is parenchymal cell necrosis that represents the cause of the high morbidity and mortality, so that new potential therapeutic approaches are indispensable for the treatment of patients at high risk of complications. However, not all factors that determine the onset and course of the disease have been explained. Aim of this article is to review the role of mitogen-activated protein kinases in pathogenesis of acute pancreatitis.展开更多
OBJECTIVE:To investigate the effect of quercetin,oleanolic acid,icariin and their compatibility on the apoptosisofhippocampalneuronsof Sprague-Dawley(SD)rats cultured with high glucose medium and the possible mechanis...OBJECTIVE:To investigate the effect of quercetin,oleanolic acid,icariin and their compatibility on the apoptosisofhippocampalneuronsof Sprague-Dawley(SD)rats cultured with high glucose medium and the possible mechanism.METHODS:The extracts were purchased from China Food and Drug Control Institute and Sellect.Hippocampus was obtained from newborn 24 h SD rats.After culturing the hippocampus in different medium for 72 h,flow cytometry was used to detect the apoptosis of hippocampal neurons,and Western blot was utilized to test the expressions of p-p38,p38,p-c-Jun N-terminal kinase(JNK)and JNK.RESULTS:Compared with the control group(CG),the neuronal apoptosis rate and the ratios of p-p38/p38 and p-JNK/JNK were significantly increased in the high glucose group(GG)(P<0.01);Compared with the GG,the apoptosis rate and the ratios ofp-p38/p38 and p-JNK/JNK were significantly decreased in other drug groups(P<0.01);Compared with the monomer groups respectively,the apoptosis rate and the ratios of p-p38/p38 and p-JNK/JNK in the two-drug groups and the three-drug group all decreased(P<0.01);Compared with the two-drug groups,the neuronal apoptosis rate and the ratio of p-JNK/JNK of the three-drug group decreased(P<0.05).CONCLUSION:Under the condition of high glucose,the quercetin,oleanolic acid and icariin can alleviate the apoptosis of hippocampus neurons,reduce the phosphorylation of p38 and JNK in p38 mitogen-activated protein kinases and JNK signaling pathway.And the efficacy of the three drugs in combination with each other can be strengthened.展开更多
OBJECTIVE:To investigate the efficacy of needling acupoins of Guanyuan(CV4),Sanyinjiao(SP6),Zusanli(ST36),Pishu(BL20),Shenshu(BL23),Zigong(EX-CA1)on the expression of p38 mitogen-activated protein kinase(p38MAPK)in ov...OBJECTIVE:To investigate the efficacy of needling acupoins of Guanyuan(CV4),Sanyinjiao(SP6),Zusanli(ST36),Pishu(BL20),Shenshu(BL23),Zigong(EX-CA1)on the expression of p38 mitogen-activated protein kinase(p38MAPK)in ovarian tissue in rats with premature ovarian failure induced by cyclophosphamide,and to study the underlying mechanism.METHODS:Forty specific pathogen free female Sprague-Dawley rats were randomly divided into the blank group,the model group,the acupuncture group,the Western Medicine group and the Western Medicine combined with acupuncture group.Except the blank group,the rest of the rats were given with cyclophosphamide for 14 d to establish premature ovarian failure model.No intervention was conducted in the blank group and model group;the acupuncture group was given with acupuncture daily;the Western Medicine group was given with estradiol valerate(0.09 mg/kg)by intragastrical gavage daily;the combination group was given with acupuncture combined with estradiol valerate(0.09 mg/kg)daily.Each group was intervened in continuously for 14 d.After the last treatment,the levels of estradiol(E2),follicle stimulating hormone(FSH)and luteinizing hormone(LH)were detected by enzyme-linked immunosorbent assay,then the ovarian tissue was dissected.Western blot was used to detect the expression of p38 MAPK protein.RESULTS:Compared with the blank group,E2 in the serum of rats in the model group was significantly decreased(P<0.05),FSH and LH were significantly increased(P<0.05),and the expression of p38 MAPK protein in the ovarian tissue of the rats was significantly increased(P<0.05).Compared with the model group,E2 in the serum of the acupuncture group,Western Medicine group and the combination group were significantly increased(P<0.05),FSH and LH levels were significantly decreased(P<0.05),and the expression of p38MAPK protein in the ovarian tissue of the rats was significantly decreased(P<0.05).However,there was no statistically significant difference between the Western Medicine group and the acupuncture group(P>0.05).CONCLUSIONS:Acupuncture has the same effect as estrogen in interfering POF caused by cyclophosphamide,and its mechanism may be related to inhibiting the expression of p38MAPK protein in ovarian tissue and affecting the activation of p38MAPK signaling pathway.展开更多
OBJECTIVE: To investigate the effect of Shenfuqiangxin capsule and the underlying mechanism on cardio-renal syndrome(CRS) in rats induced by infrarenal aortic-clamping after renal ischaemia.METHODS: Male Wistar rats u...OBJECTIVE: To investigate the effect of Shenfuqiangxin capsule and the underlying mechanism on cardio-renal syndrome(CRS) in rats induced by infrarenal aortic-clamping after renal ischaemia.METHODS: Male Wistar rats underwent infrarenal aortic-clamping after renal ischaemia or sham operation. The surviving CRS rats were divided randomly into three groups: CRS group(CRS + 10 m L·kg-1·d-1pure water by gavage), SFQX group(CRS + 13.2 g crude drug·kg- 1·d- 1Shenfuqiangxin by gavage),and handleregionpeptide(HRP)group(CRS+10mg/kg HRP by vein). Sham operation rats were given10 m L·kg-1·d-1pure water. Treatments were given8 weeks after surgery, which lasted for 4 weeks. Therats were detected for heart structure and function by transthoracic echocardiography. PPR m RNA was detected by Reverse Transcription Polymerase Chain Reaction(RT-PCR). To determine whether the mitogen-activated protein kinase(MAPK) signal pathway is included in the heart and kidney protective function of Shenfuqiangxin capsule, MAPK related proteins such as posophorylated C-Jun amino terminal kinase(p-JNK), posophorylated extracellular signal-regulated kinase ?(p-ERK1/2), posophorylated p38(p-p38) were examined by Western Blot.Apoptosis in heart and kidney tissues were detected by d UTP Nick End Labeling staining.RESULTS: Shenfuqiangxin capsule alleviated myocardial apoptosis and inhibited PRR m RNA expression and p-JNK, p-ERK1/2, p-p38 proteins expression in CRS rats.CONCLUSION: All the results suggest that Shenfuqiangxin capsule improves the injured heart and kidney function maybe through inhibition of MAPK response pathway.展开更多
To investigate the protective effect of retinoic acid （RA） on hyperoxic lung injury and the role of RA as a modulator on mitogen-activated protein kinases （MAPKs）, gastation 21 d Sprague- Dawley （SD） fetuses （t...To investigate the protective effect of retinoic acid （RA） on hyperoxic lung injury and the role of RA as a modulator on mitogen-activated protein kinases （MAPKs）, gastation 21 d Sprague- Dawley （SD） fetuses （term = 22 d） were delivered by hysterotomy. Within 12-24 h of birth, premature rat pups were randomly divided into 4 groups （n= 12 each） ： air-exposed control group （group Ⅰ ） ; hyperoxia-exposed group （ group Ⅱ ）, air-exposed plus RA group （group Ⅲ ）, hyperoxia-exposed plus RA group （group Ⅳ）. Group Ⅰ , Ⅲ were kept in room air, and group Ⅱ , Ⅳ were placed in 85 % oxygen. The pups in groups Ⅲ and Ⅳ were intraperitoneally injected with RA （500 μg/kg every day）. All lung tissues of premature rat pups were collected at the 4th day after birth. Terminal transferase d-UTP nick end labeling （TUNEL） staining was used for the detection of cell apoptosis. The expression of PCNA was immunohistochemically detected. Western blot analysis was employed for the determination of phosphorylated and total nonphosphorylated ERKs, JNKs or p38. Our results showed that lungs from the pups exposed to hyperoxia for 4 d exhibited TUNEL-positive nuclei increased markedly throughout the parenchyma （P〈0.01）, and decreased significantly after RA treatment （P〈0.01）. The index of PCNA-positive cells was significantly decreased （P〈0.01）, and was significantly increased by RA treatment （P〈0.01）. The air-space size was significantly enlarged, secondary crests were markedly decreased in hyperoxia-exposed animals. RA treatment improved lung air spaces and secondary crests in air-exposed pups, hut had no effect on hyperoxia-exposure pups. Western blotting showed that the amounts of JNK, p38 and ERK proteins in hyperoxia-exposure or RA-treated lung tissues were same as those in untreated lung tissues （P〈0.05）, whereas activation of these MAPKs was markedly altered by hyperoxia and RA. After hyperoxia exposure, p-ERK1/2, p-JNK1/2 and p-p38 were dramatically increased （P〈0.01）, whereas p-JNK1/2 and p-p38 were markedly declined and p-ERK1/2 was further elevated by RA treatment （P〈0.01）. It is concluded that RA could decrease cell apoptosis and stimulate cell proliferation under hyperoxic condition. The protection Of RA on hyperoxia-induced lung injury was related＇to the regulation of MAP kinase activation.展开更多
基金Science and Technology Development Project of Jilin Province:Preparation and Evaluation of an Animal Model of Liverdepression Type Depression (20220505038ZP)Exploring the Material Basis and Action Pathway of the Antipyretic Effect of Baihu Tang based on Histologic Techniques (20240602036RC)。
文摘OBJECTIVE:To clarify the effect of Hamayou(Oviductus Ranae) protein hydrolysate(ORPH) on depression and its exact underlying mechanism from a new perspective. METHODS:We used the Chronic Unpredictable Mild Stress(CUMS) method to prepare a mouse model of depression and lipopolysaccharide(LPS) to prepare a model of BV2 cellular inflammation to investigate the antidepressant effect and mechanism of action of ORPH. Behavioral changes in mice and cerebral blood flow were detected by behavioral experiments and scatter imaging. Levels of corticosterone(CORT), proinflammatory cytokines and neurotransmitter were detected by enzyme-linked immunosorbent assay. Furthermore, hematoxylin-eosin staining, Tunel staining were used to evaluate the effect of ORPH. The distribution and expression of ionized calcium bindingadaptor molecule-1(Iba-1) in mouse hippocampal tissue and BV2 cells were detected by immunofluorescence. Mitogen-activated protein kinase(MAPK) pathway related protein expression was detected by Western blot. RESULTS:ORPH improved depression-like behavior, ameliorated brain tissue damage and apoptosis, and inhibited microglia activation in brain tissue in mice. In addition, ORPH reduced expression of B-cell lymphoma-2(Bcl-2)-associated X(Bax), cysteinyl aspartate specific proteinase 3(Caspase3), cysteinyl aspartate specific proteinase 9(Caspase9), nuclear factor-kappa B(NF-κB), phosphorylation-p38(p-p38), phosphorylation-Jun Nterminal kinase(p-JNK) proteins, and increased expression of Bcl-2, inhibitory kappa B alpha(IκB-α), phosphorylation-extracellular regulated protein kinases 1/2(p-ERK1/2) proteins. On the other hand, there were fewer Iba-1-positive cells, lower expression of NF-κB, pp38, p-JNK and p-ERK1/2 proteins, and higher expression of IκB-α proteins in BV2 cells in the ORPH group. In addition, ORPH increased 5-hydroxytryptamine, norepinephrine levels and decreased CORT, interleukin-1β(IL-1β), interleukin-6(IL-6), tumor necrosis factor-α(TNF-α) levels. CONCLUSION:ORPH was able to improve depressionlike behaviors and that it took effects by promoting cerebral blood flow, inhibition of hypothalamic-pituitaryadrenal axis overactivation, improving the structural damage of hippocampal tissues, and inhibiting the inflammatory response. ORPH can reduced neuronal damage and inhibiting apoptosis by promoting the MAPK pathway.
基金Supported by Shanghai Putuo District Health System Science and Technology Innovation Project:Study on the Effect and Mechanism of Jinkui Shenqi Pills on Renal Water Metabolism via provirus integration site for moloney murine leukemia virus 3/aquaporin 2 Regulation (No. PTKWS202104)Chengdu University of Traditional Chinese Medicine "Xinglin Scholar" Discipline Talent Research Enhancement Plan:Based on provirus integration site for moloney murine leukemia virus 3 to Explore the Molecular Mechanism of Jingui Shenqi pill in Regulating Water Metabolism in Renal Tubular Cells (No. YYZX2022165)the Clinical Advantage Discipline of Health System of Putuo District in Shanghai (2019ysxk01)
文摘OBJECTIVE:To confirm the therapeutic effect and mechanism of Jingui Shenqi pill(金匮肾气丸,JGSQP)on cardiorenal syndrome.METHODS:Doxorubicin was used to build heart-kidney coinjury rat model.After the modeling was completed,JGSQP gavage intervention was performed.The cardiac function of rats in each group was evaluated by ultrasound detection.Serum of rats was collected and examined for markers of heart and kidney damage.Enzyme linked immunosorbent assay detected serum inflammatory factors interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)expression.Quantitative real-time polymerase chain reaction(PCR)and Western blot detected the changes of related genes and proteins.RESULTS:JGSQP significantly increased left ventricular ejection fraction(EF)and left ventricular shortening fraction(FS)values,decreased the heart and kidney damage markers and fibrosis levels(P<0.05).Furthermore,it can reduce IL-1β,IL-6,and TNF-αinflammatory expression(P<0.05).Mechanistically,JGSQP significantly inhibited the expression of key genes and proteins of mitogen-activated protein kinase(MAPK)signaling pathway(P<0.05).CONCLUSIONS:Jingui Shenqi pill can exert therapeutic effects on cardiorenal syndrome by inhibiting the activation of the MAPK signaling pathway and inflammatory responses.
文摘Objective To evaluate the antagonistic effects of N-acetylcysteine(NAC)on mitogen-activated protein kinases(MAPK)pathway activation,oxidative stress and inflammatory responses in rats with lung injury induced by fine particulate matter(PM2.5).Methods Forty eight male Wistar rats were randomly divided into six groups:blank control group(C1),water drip control group(C2),PM2.5 exposed group(P),low-dose NAC treated and PM2.5 exposed group(L),middle-dose NAC treated and PM2.5 exposed group(M),and high-dose NAC treated and PM2.5 exposed group(H).PM2.5 suspension(7.5 mg/kg)was administered tracheally once a week for four times.NAC of 125 mg/kg,250 mg/kg and 500 mg/kg was delivered intragastrically to L,M and H group respectively by gavage(10 ml/kg)for six days before PM2.5 exposure.The histopathological changes and human mucin 5 subtype AC(MUC5AC)content in lung tissue of rats were evaluated.We investigated IL-6 in serum and bronchoalveolar lavage fluid(BALF)by Enzyme-linked immunosorbent assay(ELISA),MUC5AC in lung tissue homogenate by ELISA,glutathione peroxidase(GSH-PX)in serum and BALF by spectrophotometry,and the expression of p-ERK1/2,p-JNK1/2 and p-p38 proteins by Western blot.All the measurements were analyzed and compared statistically.Results Lung tissue of rats exposed to PM2.5 showed histological destruction and increased mucus secretion of bronchial epithelial cells.Rats receiving NAC treatment showed less histological destruction and mucus secretion.Of P,L,M and H group,MUC5AC in lung tissue,IL-6 in serum and BALF were higher than controls(C1 and C2)(all P<0.05),with the highest levels found in the P group and a decreasing trend with increase of NAC dose.The activity of GSH-PX in serum and BALF of PM2.5 exposed rats(P,L,M and H)was lower than that of controls(all P<0.05),with higher activities found in NAC treated rats(L,M,and H),and an increasing trend with increase of NAC dose.The expressions of p-ERK1/2,p-JNK1/2 and p-p38 proteins in PM2.5 exposed lung tissue(P,L,M and H)was higher than controls(all P<0.05),with decreased levels and dose dependent downregulation found in NAC treated rats.Conclusion NAC can antagonize major MAPK pathway activation,lung oxidative stress and inflammatory injury induced by PM2.5 in rats.
基金financially supported by National Natural Science Foundation of China(Nos.81302794,81071841,81102853)the Study of Marsdenia tenacissima extract(MTE):Study on quality control of antitumor traditional Chinese medicine Xiao-Ai-Ping injection(No.2011ZX09201-201)
文摘Marsdenia tenacissima extract(MTE, trade name: Xiao-Ai-Ping injection) is an extract of a single Chinese plant medicine. It has been used for the treatment of cancer in China for decades, especially for esophageal cancer and other cancers in the digestive tract. In the present study, the potential mechanism for MTE's activity in esophageal cancer was explored. The effects of MTE on the proliferation of human esophageal cancer cells(KYSE150 and Eca-109) were investigated by the MTT assay, the Brd U(bromodeoxyuridine) incorporation immunofluorescence assay, and flow cytometric analysis. MTE inhibited cell proliferation through inducing G0/G1 cell cycle arrest in KYSE150 and Eca-109. Western blot analysis was employed to determine protein levels in the MTE treated cells. Compared with the control cells, the expression levels of the cell cycle regulatory proteins cyclin D1/D2/D3, cyclin E1, CDK2/4/6(CDK: cyclin dependent kinase), and p-Rb were decreased significantly in the cells treated with MTE at 40 mg·m L-1. In addition, MTE had an inhibitory effect on the MAPK(mitogen-activated protein kinase) signal transduction pathway, including ERK(extracellular signal-regulated kinase), JNK(c-Jun N-terminal kinase), and p38 MAPK. Moreover, MTE showed little additional effects on the regulation of cyclin D1/D3, CDK4/6, and p-Rb when the ERK pathway was already inhibited by the specific ERK inhibitor U0126. In conclusion, these data suggest that MTE inhibits human esophageal cancer cell proliferation through regulation of cell cycle regulatory proteins and the MAPK signaling pathways, which is probably mediated by the inhibition of ERK activation.
基金supported by the National Natural Science Foundation of China,No.81173355
文摘Electroacupuncture attenuates cerebral hypoxia and neuronal apoptosis induced by cerebral ischemia/reperfusion injury.To further identify the involved mechanisms,we assumed that electroacupuncture used to treat cerebral ischemia/reperfusion injury was associated with the p38 mitogen-activated protein kinase（MAPK） signaling pathway.We established rat models of cerebral ischemia/reperfusion injury using the modified Zea-Longa＇s method.At 30 minutes before model establishment,p38 MAPK blocker SB20358 was injected into the left lateral ventricles.At 1.5 hours after model establishment,electroacupuncture was administered at acupoints of Chize（LU5）,Hegu（LI4）,Zusanli（ST36）,and Sanyinjiao（SP6） for 20 minutes in the affected side.Results showed that the combination of EA and SB20358 injection significantly decreased neurologic impairment scores,but no significant differences were determined among different interventional groups.Hematoxylin-eosin staining also showed reduced brain tissue injuries.Compared with the SB20358 group,the cells were regularly arranged,the structures were complete,and the number of viable neurons was higher in the SB20358 ＋ electroacupuncture group.Terminal deoxynucleotidyl transferase（Td T）-mediated d UTP nick-end labeling assay showed a decreased apoptotic index in each group,with a significant decrease in the SB20358 ＋ electroacupuncture group.Immunohistochemistry revealed reduced phosphorylated p38 expression at 3 days in the electroacupuncture group and SB20358 ＋ electroacupuncture group compared with the ischemia/reperfusion group.There was no significant difference in phosphorylated p38 expression between the ischemia/reperfusion group and SB20358 group.These findings confirmed that the electroacupuncture effects on mitigating cerebral ischemia/reperfusion injury are possibly associated with the p38 MAPK signaling pathway.A time period of 3 days could promote the repair of ischemic cerebral nerves.
基金Supported by Technology Foundation of Ministry of Education, China
文摘AIM: To investigate the expression of mitogen-activated protein kinases (MAPKs) and its upstream protein kinase in human gastric cancer and to evaluate the relationship between protein levels and clinicopathological parameters. METHODS: Western blot was used to measure the expression of extracellular signal-regulated kinase (ERK)-1, ERK-2, ERK-3, p38 and mitogen or ERK activated protein kinaseMEK-1 proteins in surgically resected gastric carcinoma, adjacent normal mucosa and metastatic lymph nodes from 42 patients. Immunohistochemistry was employed for their localization. RESULTS: Compared with normal tissues, the protein levels of ERK-1 (integral optical density value 159 526?5 760 vs 122 807±65 515, P= 0.001), ERK-2 (168 471±95 051 vs 120 469±72 874, P<0.001), ERK-3 (118 651±71 513 vs 70 934±68 058,P<0.001), P38 (104 776±51 650 vs 82 930±40 392, P= 0.048) and MEK-1 (116 486±45 725 vs 101 434±49 387, P = 0.027) were increased in gastric cancer tissues. Overexpression of ERK-3 was correlated to TNM staging [average ratio of integral optic density (IOD)tumor: IODnormal in TNM I, II, III, IV tumors was 1.43±0.34, 5.08±3.74, 4.99±1.08, 1.44±1.02, n = 42, P= 0.023] and serosa invasion (4.31±4.34 vs 2.00±2.03, P = 0.037). In poorly differentiated cancers (n = 33), the protein levels of ERK-1 and ERK-2 in stage III and IV tumors were higher than those in stage I and II tumors (2.64+3.01 vs 1.01±0.33, P= 0.022; 2.05±1.54 vs1.24±0.40, P= 0.030). Gastric cancer tissues with either lymph node involvement (2.49±2.91 vs1.03±0.36, P= 0.023; 1.98±1.49vs1.24±0.44, P= 0.036) or serosa invasion (2.39±2.82 vs 1.01±0.35, P= 0.022; 1.95±1.44 vs1.14±0.36, P=0.015) expressed higher protein levels of ERK-1 and ERK-2. In Borrmann II tumors, expression of ERK-2 and ERK-3 was increased compared with Borrmann III tumors (2.57±1.86 vs1.23±0.60, P= 0.022; 5.50±5.05 vs1.83±1.21, P= 0.014). Borrmann IV tumors expressed higher p38 protein levels. No statistically significant difference in expression of MAPKs was found when stratified to tumor size or histological grade (P>0.05). Protein levels of ERK-2, ERK-3 and MEK-1 in metastatic lymph nodes were 2-7 folds higher than those in adjacent normal mucosa. The immunohistochemistry demonstrated that ERK-1, ERK-2, ERK-3, p38 and MEK-1 proteins were mainly localized in cytoplasm. The expression of MEK-1 in gastric cancer cells metastasized to lymph nodes was higher than that of the primary site. CONCLUSION: MAPKs, particularly ERK subclass are overexpressed in the majority of gastric cancers. Overexpression of ERKs is correlated to TNM staging, serosa invasion, and lymph node involvement. The overexpression of p38 most likely plays a prominent role in certain morphological subtypes of gastric cancers. MEK-1 is also overexpressed in gastric cancer, particularly in metastatic lymph nodes. Upregulation of MARK signal transduction pathways may play an important role in tumorigenesis and metastatic potential of gastric cancer.
文摘Cold-inducible RNA-binding protein （CIRP） is an RNA-binding protein that is expressed in normal testes and downregulated after heat stress caused by cryptorchidism, varicocele or environmental temperatures. The purpose of this study was to investigate the functions of CIRP in the testes. We employed RNAi technique to knock down the expression of CIRP in the testes, and performed haematoxylin and eosin staining to evaluate morphological changes following knockdown. Germ cell apoptosis was examined by terminal deoxynucleotidal transferase-mediated dUTP nick end labelling （TUNEL） assay, and mitogen-activated protein kinase （MAPK） signalling pathways were investigated by Western blotting to determine the possible mechanism of apoptosis. We found that using siRNA is a feasible and reliable method for knocking down gene expression in the testes. Compared to controls, the mean seminiferous tubule diameter （MSTD） and the thickness of the germ cell layers decreased following siRNA treatment, whereas the percentage of apoptotic seminiferous tubules increased. The p44/p42, p38 and SAPK/JNK MAPK pathways were activated after downregulation of CIRP. In conclusion, we discovered that downregulation of CIRP resulted in increased germ cell apoptosis, possibly viathe activation of the p44/p42, p38 and SAPK/JNK MAPK pathways.
文摘OBJECTIVE: To explore the effects of evodiamine on ovarian cancer cells and the mechanisms underlying such effects.METHODS: Human ovarian cancer cells HO-8910 PM were treated with evodiamine at 0, 1.25,2.5, and 5 μM for 1-4 d. 3-(4,5-Dimethiylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay was used to detect the growth inhibition rate of evodiamine-treated HO-8910 PM cells. The cell cycle was observed via propidium iodide(PI) staining. Apoptosis induction was assessed via Annexin V-fluorescein isothiocyanate/propidium iodide(Annexin V-FITC/PI) double staining assay. To verify the mechanism of apoptosis, caspase-dependent apoptotic pathway-related protein was detected by Western blot analysis. The expression levels of mitogen-activated protein kinase(MAPK)and/or phosphatidylinositol-3-kinase(PI3K)/pro-tein kinase B(Akt) pathway-related proteins were also investigated.RESULTS: Evodiamine significantly inhibited the proliferation of HO-8910 PM cells in a dose- and time-dependent manner. Evodiamine induced G2/M arrest with an increase of cyclin B1 level, and promoted cell apoptosis with a decrease of B cell lymphoma/lewkmia-2(Bcl-2) and an increase of Bcl-2-associated X protein(Bax) level. In addition,evodiamine treatment led to the activation of caspase-8, caspase-9, and caspase-3 and the cleavage of poly(ADP-ribose)-polymerase(PARP). Evodiamine targeted the MAPK and/or PI3K/Akt pathways by reducing the expression and activity of PI3 K, Akt, and extracellular signal-regulated kinase mitogen-activated protein kinase(ERK1/2 MAPK)and the activity of p38 MAPK.CONCLUSION: Evodiamine can inhibit the growth of ovarian cancer cells by G2/M arrest and intrinsic and extrinsic apoptosis. In addition, evodiamine-induced PI3K/Akt, ERK1/2 MAPK, and p38 MAPK signaling may be involved in cell death.
基金Supported by the National Basic Science and Development Programme (973 Programme),No.G1999054204 National Natural Science Foundation of China, No. 30170966, 30230370 National High-Technology Programme (863 Programme), No. 2001AA215131
文摘AIM: To investigate the role of p38 mitogen-activated protein kinase in rat small intestine after ischemia-reperfusion (I/R)insult and the relationship between activation of p38 MAPK and apoptotic cell death of intestine.METHODS: Ninety Wistar rats were divided randomly into three groups, namely sham-operated group (C), I/R vehicle group (R) and SB203580 pre-treated group(S).In groups R and S, the superior mesenteric artery(SMA)was separated and occluded for 45 min, then released for reperfusion for0.25, 0.5, 1, 2, 6, 12 and 24 h. In group C, SMA was separated without occlusion. Plasma D-lactate levels were examined and histological changes were observed under a light microscope. The activity of p38 MAPK was determined by Western immunoblotting and apoptotic cells were detected by the terminal deoxynucleotidyl transferase (TdT)-mediated dUDP-biotin nick end labeling (TUNEL).RESULTS: Intestinal ischemia followed by reperfusion activated p38 MAPK, and the maximal level of activation (7.3-fold vs sham-operated group) was reached 30 min after I/R. Treatment with SB 203580, a p38 MAPK inhibitor,reduced intestinal apoptosis (26.72±3.39% vs62.50±3.08%in I/R vehicle, P＜0.01) and decreased plasma D-lactate level (0.78±0.15 mmol/L in I/R vehicle vs0.42±0.17 mmol/L in SB-treated group) and improved post-ischemic intestinal histological damage.CONCLUSION: p38 MAPK plays a crucial role in the signal transduction pathway mediating post-ischemic intestinal apoptosis, and inhibition of p38 MAPK may attenuate ischemia-reperfusion injury.
文摘The mitogen-activated protein kinase（MAPK） signaling pathway plays an important role in the regulation of cell growth, proliferation, differentiation, transformation and death. Mitogen-activated protein kinase phosphatase 1（MKP1） has an inhibitory effect on the p38 MAPK and JNK pathways, but it is unknown whether it plays a role in Aβ-induced oxidative stress and neuronal inflammation. In this study, PC12 cells were infected with MKP1 sh RNA, MKP1 lentivirus or control lentivirus for 12 hours, and then treated with 0.1, 1, 10 or 100 μM amyloid beta 42（Aβ42）. The cell survival rate was measured using the cell counting kit-8 assay. MKP1, tumor necrosis factor-alpha（TNF-α） and interleukin-1β（IL-1β） m RNA expression levels were analyzed using quantitative real time-polymerase chain reaction. MKP1 and phospho-c-Jun N-terminal kinase（JNK） expression levels were assessed using western blot assay. Reactive oxygen species（ROS） levels were detected using 2′,7′-dichlorofluorescein diacetate. Mitochondrial membrane potential was measured using flow cytometry. Superoxide dismutase activity and malondialdehyde levels were evaluated using the colorimetric method. Lactate dehydrogenase activity was measured using a microplate reader. Caspase-3 expression levels were assessed by enzyme-linked immunosorbent assay. Apoptosis was evaluated using the terminal deoxynucleotidyl transferase d UTP nick end labeling method. MKP1 overexpression inhibited Aβ-induced JNK phosphorylation and the increase in ROS levels. It also suppressed the Aβ-induced increase in TNF-α and IL-1β levels as well as apoptosis in PC12 cells. In contrast, MKP1 knockdown by RNA interference aggravated Aβ-induced oxidative stress, inflammation and cell damage in PC12 cells. Furthermore, the JNK-specific inhibitor SP600125 abolished this effect of MKP1 knockdown on Aβ-induced neurotoxicity. Collectively, these results show that MKP1 mitigates Aβ-induced apoptosis, oxidative stress and neuroinflammation by inhibiting the JNK signaling pathway, thereby playing a neuroprotective role.
基金Supported by Natural Science Foundation-funded Project:Pingchuan Formula Regulates the Effect of Pi3k/Akt Pathway on Airway Inflammation and Airway Remodeling in Asthma and its Mechanism(No.81603662)Shanghai Municipal Commission of Health and Family Planning:"Shanghai School of Traditional Chinese Medicine"Xu’s Pediatric Diagnosis and Treatment[Center Construction ZY(2018-2020)-CCCX-1012]。
文摘OBJECTIVE:To examine the role and decipher the mechanism of Pingchuan formula(平喘方,PCF)in treating allergic asthma.METHODS:The mice were treated with saline,dexamethasone(DXM)and PCF for 1 week after the asthma model was established and their respiratory function including respiratory resistance(RI),pulmonary dynamic compliance(Cdyn)and maximum voluntary ventilation(MVV)were measured.In addition,cellular changes in bronchoalveolar lavage fluid(BALF)and pathological changes in lung biopsy as well as the expression level ofα-smooth muscle actin(α-SMA),transforming growth factor-beta1(TGF-β1)in BALF and interleukin-5(IL-5),interleukin-13(IL-13),tumor necrosis factor-α(TNF-α),interferon-γ(IFN-γ),nuclear factor-kappa B-p65(NF-κBp65),inhibitor-αof nuclear transcription factorκB(IκBα),p38 mitogen-activated protein kinase(p38 MAPK),c-jun n-terminal kinase(JNK)and its phosphorylated proteins in lung tissue were also examined and compared among different groups.RESULTS:Our data suggested that the respiratory functions were significantly improved and the pathological changes ameliorated in the DXM group and the PCF group compared to the model group.Both DXM and PCF effectively decreased the number of eosinophils,lymphocytes,and neutrophils in BAL as well as the secretion ofα-SMA and TGF-β1,IL-5,IL-13,while increased the expression of TNF-αand IFN-γ.Furthermore,our study indicated that the NF-κBp65,IκBα,p38 MAPK and JNK pathways were inhibited under the treatment of PCF.CONCLUSION:Our data indicated that PCF can attenuate the inflammatory response in asthma through inhibiting the NF-κB/MAPK signaling pathway.This study not only supported the use of PCF in allergic asthma in clinic but also shed light upon afurther understanding of the disease pathogenesis.
基金Supported by the National Natural Science Foundation of China, No. 30340036 and No. 30470891 Grant from Jiangsu University and Zhenjiang Key Institute of Clinical Laboratory Medicine (SH2006066)
文摘AIM: To explore the mechanism by which H pylori causes activation of gastric epithelial cells. METHODS: A VacA (+) and CagA (+) standard H pylori line NCTC 11637 and a human gastric adenocarcinoma derived gastric epithelial cell line BGC-823 were applied in the study. MTT assay and 3H-TdR incorporation test were used to detect the proliferation of BGC-823 cells and Western blotting was used to detect the activity and existence of related proteins. RESULTS: Incubation with H pylori extract increased the proliferation of gastric epithelial cells, reflected by both live cell number and DNA synthesis rate. The activity of extracellular signal-regulated protein kinase (ERK) signal transduction cascade increased within 20 min after in- cubation with H pylori extract and appeared to be a sus- tained event. MAPK/ERK kinase (MEK) inhibitor PD98059 abolished the action of H pylori extract on both ERK activity and cell proliferation. Incubation with H pylori extract increased c-Fos expression and SRE-dependent gene expression. H pylori extract caused phosphorylation of several proteins including a protein with molecular size of 97.4 kDa and tyrosine kinase inhibitor genistein inhibited the activation of ERK and the proliferation of cells caused by H pylori extract. CONCLUSION: Biologically active elements in H pylori extract cause proliferation of gastric epithelial cells through activating tyrosine kinase and ERK signal trans- duction cascade.
基金This work was supported by the National Natural Science Foundation of China(82172182 and 82102311)Social Development Projects of Jiangsu Province(BE2017720)+2 种基金Natural Science Foundation of Jiangsu Province(BK20190247)Science Foundation of Jiangsu Health Commission(H2018039)Jiangsu Postdoctoral Research Foundation(2018K048A and 2020Z193).
文摘BACKGROUND:Paraquat(PQ)-induced acute lung injury(ALI)and pulmonary fi brosis are common diseases with high mortality but without eff ective antidotes in emergency medicine.Our previous study has proved that arctigenin suppressed pulmonary fibrosis induced by PQ.We wondered whether arctigenin could also have a protective eff ect on PQ-induced ALI.METHODS:A PQ-induced A549 cell injury model was used,and the effect of arctigenin was determined by a cell counting kit-8(CCK-8)cell viability assay.In addition,terminal deoxynucleotidyl transferase(TdT)-mediated dUTP nick-end labelling(TUNEL)staining assays and mitochondrial membrane potential assays were performed to evaluate the level of cell apoptosis.The generation of reactive oxygen species(ROS)was refl ected by dihydroethidium(DHE)staining and a 2’,7’-dichlorodihy drofluorescein diacetate(DCFH-DA)assay.Moreover,immunoblotting studies were used to assess the expression of mitogen-activated protein kinases(MAPKs)and p38 MAPK.RESULTS:Arctigenin attenuated PQ-induced inhibition of A549 cell viability in a dose-dependent manner.Arctigenin also significantly reduced PQ-induced A549 cell apoptosis,as refl ected by the TUNEL assay and mitochondrial membrane potential assay,which may result from suppressed ROS/p38 MAPK signaling because we found that arctigenin dramatically suppressed ROS generation and p38 MAPK phosphorylation.CONCLUSION:Arctigenin could attenuate PQ-induced lung epithelial A549 cell injury in vitro by suppressing ROS/p38 MAPK-mediated cell apoptosis,and arctigenin might be considered a potential candidate drug for PQ-induced ALI.
基金Supported by China Postdoctoral Science Foundation (Experimental Study on the Inhibitory Effect of Yiqi Tongluo Method on RGCs Apoptosis in AION Model Rats, No. 2017M621180)。
文摘OBJECTIVE: To study the molecular mechanism of Buyang Huanwu Tang( 补阳还五汤)(BHT) protecting retinal ganglion cells(RGCs) from oxygen induced oxidative stress and apoptosis after anterior ischemia. METHODS: In this study, the Chinese herbs of BHT were extracted by first boiling in water, then were filtered, concentrated, and freeze-dried. The chemical profile of BHT extract was determined by liquid chromatography mass spectrometry(LC-MS). H2O2-induced RGC-5 cells were used as a cell model to investigate the protective effect and mechanism of BHT on RGCs. RESULTS: The survival rate of damaged RGC-5 by BHT was significantly increased by the 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazolium-romid method. Fluorescence activating cell sorter(FACS analysis) showed that BHT could significantly reduce apoptosis induced by oxidative stress via the reactive oxygen species(ROS)-mitogen-activated protein kinase(MAPK)-Caspase-3 signal pathway. CONCLUSION: BHT possesses a high antioxidant capacity and could significantly reduce ROS levels of RGC-5 cells damaged by H2O2.Therefore, the present study has provided possible alternative strategies for the prevention and treatment of ischemic optic disease by using traditional Chinese herbal formulas.
基金Supported by National Natural Science Foundation of China,No.81472208the Open Projects of State Key Laboratory of Molecular Oncology,No.SKL-KF-2015-12
文摘The mortality rate of gastric cancer worldwide is as high as 70%, despite the development of novel therapeutic strategies. One reason for the high mortality is the rapid and uninhibited spread of the disease, such that the majority of patients are diagnosed at a stage when efficient therapeutic treatment is not available. Therefore, in-depth research is needed to investigate the mechanism of gastric cancer metastasis and invasion to improve outcomes and provide biomarkers for early diagnosis. The mitogen-activated protein kinase(MAPK) signaling pathway is widely expressed in multicellular organisms, with critical roles in multiple biological processes, such as cell proliferation, death, differentiation, migration, and invasion. The MAPK pathway typically responds to extracellular stimulation. However, the MAPK pathway is often involved in the occurrence and progression of cancer when abnormally regulated. Many studies have researched the relationship between the MAPK signaling pathway and cancer metastasis and invasion, but little is known about the important roles that the MAPK signaling pathway plays in gastric cancer. Based on an analysis of published data, this review aims to summarize the important role that the MAP kinases play in the invasion and metastasis of gastric cancer and attempts to provide potential directions for further research and clinical treatment.
文摘Acute pancreatitis(AP) is an inflammatory disease characterized by acute inflammation and necrosis of the pancreatic parenchyma. AP is often associated with organ failure, sepsis, and high mortality. The pathogenesis of AP is still not well understood. In recent years several papers have highlighted the cellular and molecular events of acute pancreatitis. Pancreatitis is initiated by activation of digestive enzymes within the acinar cells that are involved in autodigestion of the gland, followed by a massive infiltration of neutrophils and macrophages and release of inflammatory mediators, responsible for the local and systemic inflammatory response. The hallmark of AP is parenchymal cell necrosis that represents the cause of the high morbidity and mortality, so that new potential therapeutic approaches are indispensable for the treatment of patients at high risk of complications. However, not all factors that determine the onset and course of the disease have been explained. Aim of this article is to review the role of mitogen-activated protein kinases in pathogenesis of acute pancreatitis.
文摘OBJECTIVE:To investigate the effect of quercetin,oleanolic acid,icariin and their compatibility on the apoptosisofhippocampalneuronsof Sprague-Dawley(SD)rats cultured with high glucose medium and the possible mechanism.METHODS:The extracts were purchased from China Food and Drug Control Institute and Sellect.Hippocampus was obtained from newborn 24 h SD rats.After culturing the hippocampus in different medium for 72 h,flow cytometry was used to detect the apoptosis of hippocampal neurons,and Western blot was utilized to test the expressions of p-p38,p38,p-c-Jun N-terminal kinase(JNK)and JNK.RESULTS:Compared with the control group(CG),the neuronal apoptosis rate and the ratios of p-p38/p38 and p-JNK/JNK were significantly increased in the high glucose group(GG)(P<0.01);Compared with the GG,the apoptosis rate and the ratios ofp-p38/p38 and p-JNK/JNK were significantly decreased in other drug groups(P<0.01);Compared with the monomer groups respectively,the apoptosis rate and the ratios of p-p38/p38 and p-JNK/JNK in the two-drug groups and the three-drug group all decreased(P<0.01);Compared with the two-drug groups,the neuronal apoptosis rate and the ratio of p-JNK/JNK of the three-drug group decreased(P<0.05).CONCLUSION:Under the condition of high glucose,the quercetin,oleanolic acid and icariin can alleviate the apoptosis of hippocampus neurons,reduce the phosphorylation of p38 and JNK in p38 mitogen-activated protein kinases and JNK signaling pathway.And the efficacy of the three drugs in combination with each other can be strengthened.
文摘OBJECTIVE:To investigate the efficacy of needling acupoins of Guanyuan(CV4),Sanyinjiao(SP6),Zusanli(ST36),Pishu(BL20),Shenshu(BL23),Zigong(EX-CA1)on the expression of p38 mitogen-activated protein kinase(p38MAPK)in ovarian tissue in rats with premature ovarian failure induced by cyclophosphamide,and to study the underlying mechanism.METHODS:Forty specific pathogen free female Sprague-Dawley rats were randomly divided into the blank group,the model group,the acupuncture group,the Western Medicine group and the Western Medicine combined with acupuncture group.Except the blank group,the rest of the rats were given with cyclophosphamide for 14 d to establish premature ovarian failure model.No intervention was conducted in the blank group and model group;the acupuncture group was given with acupuncture daily;the Western Medicine group was given with estradiol valerate(0.09 mg/kg)by intragastrical gavage daily;the combination group was given with acupuncture combined with estradiol valerate(0.09 mg/kg)daily.Each group was intervened in continuously for 14 d.After the last treatment,the levels of estradiol(E2),follicle stimulating hormone(FSH)and luteinizing hormone(LH)were detected by enzyme-linked immunosorbent assay,then the ovarian tissue was dissected.Western blot was used to detect the expression of p38 MAPK protein.RESULTS:Compared with the blank group,E2 in the serum of rats in the model group was significantly decreased(P<0.05),FSH and LH were significantly increased(P<0.05),and the expression of p38 MAPK protein in the ovarian tissue of the rats was significantly increased(P<0.05).Compared with the model group,E2 in the serum of the acupuncture group,Western Medicine group and the combination group were significantly increased(P<0.05),FSH and LH levels were significantly decreased(P<0.05),and the expression of p38MAPK protein in the ovarian tissue of the rats was significantly decreased(P<0.05).However,there was no statistically significant difference between the Western Medicine group and the acupuncture group(P>0.05).CONCLUSIONS:Acupuncture has the same effect as estrogen in interfering POF caused by cyclophosphamide,and its mechanism may be related to inhibiting the expression of p38MAPK protein in ovarian tissue and affecting the activation of p38MAPK signaling pathway.
基金Natural Science Foundation-funded Project:Study on Mechanism of Invigorate Qi Warm Yang Activate Blood And Promote Dieresis Treatment Adjusted By MAPK Mediated by(P)RR(No.30973795)Sponsored by the the National Natural Science Foundation of China(No.81202801)
文摘OBJECTIVE: To investigate the effect of Shenfuqiangxin capsule and the underlying mechanism on cardio-renal syndrome(CRS) in rats induced by infrarenal aortic-clamping after renal ischaemia.METHODS: Male Wistar rats underwent infrarenal aortic-clamping after renal ischaemia or sham operation. The surviving CRS rats were divided randomly into three groups: CRS group(CRS + 10 m L·kg-1·d-1pure water by gavage), SFQX group(CRS + 13.2 g crude drug·kg- 1·d- 1Shenfuqiangxin by gavage),and handleregionpeptide(HRP)group(CRS+10mg/kg HRP by vein). Sham operation rats were given10 m L·kg-1·d-1pure water. Treatments were given8 weeks after surgery, which lasted for 4 weeks. Therats were detected for heart structure and function by transthoracic echocardiography. PPR m RNA was detected by Reverse Transcription Polymerase Chain Reaction(RT-PCR). To determine whether the mitogen-activated protein kinase(MAPK) signal pathway is included in the heart and kidney protective function of Shenfuqiangxin capsule, MAPK related proteins such as posophorylated C-Jun amino terminal kinase(p-JNK), posophorylated extracellular signal-regulated kinase ?(p-ERK1/2), posophorylated p38(p-p38) were examined by Western Blot.Apoptosis in heart and kidney tissues were detected by d UTP Nick End Labeling staining.RESULTS: Shenfuqiangxin capsule alleviated myocardial apoptosis and inhibited PRR m RNA expression and p-JNK, p-ERK1/2, p-p38 proteins expression in CRS rats.CONCLUSION: All the results suggest that Shenfuqiangxin capsule improves the injured heart and kidney function maybe through inhibition of MAPK response pathway.
基金This project was supported by a grant from the NationalKey Science and Technology Program of the Tenth Five-years-Plan (No .2004BA720A11) ,and a grant from Nation-al Natural Sciences Foundation of China (No .30471824)
文摘To investigate the protective effect of retinoic acid （RA） on hyperoxic lung injury and the role of RA as a modulator on mitogen-activated protein kinases （MAPKs）, gastation 21 d Sprague- Dawley （SD） fetuses （term = 22 d） were delivered by hysterotomy. Within 12-24 h of birth, premature rat pups were randomly divided into 4 groups （n= 12 each） ： air-exposed control group （group Ⅰ ） ; hyperoxia-exposed group （ group Ⅱ ）, air-exposed plus RA group （group Ⅲ ）, hyperoxia-exposed plus RA group （group Ⅳ）. Group Ⅰ , Ⅲ were kept in room air, and group Ⅱ , Ⅳ were placed in 85 % oxygen. The pups in groups Ⅲ and Ⅳ were intraperitoneally injected with RA （500 μg/kg every day）. All lung tissues of premature rat pups were collected at the 4th day after birth. Terminal transferase d-UTP nick end labeling （TUNEL） staining was used for the detection of cell apoptosis. The expression of PCNA was immunohistochemically detected. Western blot analysis was employed for the determination of phosphorylated and total nonphosphorylated ERKs, JNKs or p38. Our results showed that lungs from the pups exposed to hyperoxia for 4 d exhibited TUNEL-positive nuclei increased markedly throughout the parenchyma （P〈0.01）, and decreased significantly after RA treatment （P〈0.01）. The index of PCNA-positive cells was significantly decreased （P〈0.01）, and was significantly increased by RA treatment （P〈0.01）. The air-space size was significantly enlarged, secondary crests were markedly decreased in hyperoxia-exposed animals. RA treatment improved lung air spaces and secondary crests in air-exposed pups, hut had no effect on hyperoxia-exposure pups. Western blotting showed that the amounts of JNK, p38 and ERK proteins in hyperoxia-exposure or RA-treated lung tissues were same as those in untreated lung tissues （P〈0.05）, whereas activation of these MAPKs was markedly altered by hyperoxia and RA. After hyperoxia exposure, p-ERK1/2, p-JNK1/2 and p-p38 were dramatically increased （P〈0.01）, whereas p-JNK1/2 and p-p38 were markedly declined and p-ERK1/2 was further elevated by RA treatment （P〈0.01）. It is concluded that RA could decrease cell apoptosis and stimulate cell proliferation under hyperoxic condition. The protection Of RA on hyperoxia-induced lung injury was related＇to the regulation of MAP kinase activation.
