Objective:To investigate the synergistic effects of auranofin and schisandrin A(SA)on cell proliferation inhibition and apoptosis induction in human hepatocellular carcinoma Hep3B cells.Methods:Cell viability was asse...Objective:To investigate the synergistic effects of auranofin and schisandrin A(SA)on cell proliferation inhibition and apoptosis induction in human hepatocellular carcinoma Hep3B cells.Methods:Cell viability was assessed using MTT to determine the synergistic effects of auranofin and SA.Three-dimensional(3D)culture models were used to evaluate the effects on spheroid structure and size.Apoptosis was analyzed by flow cytometry for sub-G1 populations,annexin V staining,and Western blotting for apoptotic markers.Reactive oxygen species(ROS)production was measured using DCF-DA staining.Results:Our results showed that combined treatment with auranofin and SA led to a significant reduction in cell viability compared with either compound alone,with isobologram analysis confirming their synergistic interactions.Under 3D culture conditions,auranofin and SA disrupted the compact structure of spheroids,leading to a loosened and disorganized morphology at the periphery,which appeared as an increase in spheroid size.Moreover,the induction of apoptosis by auranofin and SA was evidenced by elevated sub-G1 phase populations,increased annexin V-positive cells,and upregulation of apoptotic markers such as cleaved poly(ADPribose)polymerase 1 and cleaved caspase-3.Notably,auranofin combined with SA markedly enhanced ROS production,which was mitigated by the ROS scavenger N-acetylcysteine.Additionally,the phosphoinositide 3-kinase(PI3K)/Akt signaling pathway was downregulated in response to auranofin and SA treatment,and further apoptotic effects were observed following PI3K inhibition with LY294002.Conclusions:Auranofin combined with SA promotes apoptosis of hepatocellular carcinoma via ROS generation and inhibition of the PI3K/Akt pathway.展开更多
AIM： To illuminate the molecular targets for schisandrin against cerebrovascular disease based on the combined methods of network pharmacology prediction and experimental verification. METHOD： A protein database was...AIM： To illuminate the molecular targets for schisandrin against cerebrovascular disease based on the combined methods of network pharmacology prediction and experimental verification. METHOD： A protein database was established through constructing the drug-protein network from literature mining data. The protein-protein network was built through an in-depth exploration of the relationships between the proteins. The computational platform was implemented to predict and extract the sensitive sub-network with significant P-values from the protein-protein network. Then the key targets and pathways were identified from the sensitive sub-network. The most related targets and pathways were also confirmed in hydrogen peroxide （H202）-induced PC 12 cells by Western blotting. RESULTS： Twelve differentially expressed proteins （gene names： NFKB1, RELA, TNFSF10, MAPK1, CHUK, CASP8, PIGS2, MAPK 14, CREBI, IFNG, APR and BCL2） were confirmed as the central nodes of the interaction network （45 nodes, 93 edges）. The NF-KB signaling pathway was suggested as the most related pathway of schisandrin for cerebrovascular disease. Furthermore, schisandrin was found to suppress the expression and phosphorylation of 1KKct, as well as p50 and p65 induced by H2O2 in PC12 cells by Western blotting. CONCLUSION： The computational platform that integrates literature mining data, protein-protein interactions, sensitive sub-network, and pathway results in identification of the NF-arB signaling pathway as the key targets and pathways for schisandrin.展开更多
AIM： To investigate the inhibitory effect and possible mechanism of action of schisandrin B in SC-B on gastric cancer cells in vitro.METHODS： SC-B consisted of schisandrin B, aloeemodin, and Astragalus polysaccharid...AIM： To investigate the inhibitory effect and possible mechanism of action of schisandrin B in SC-B on gastric cancer cells in vitro.METHODS： SC-B consisted of schisandrin B, aloeemodin, and Astragalus polysaccharides. Exponentially growing human gastric cancer SGO7901 cells were divided into six treatment groups： （1） control group （RPMI 1640 medium）; （2） negative control group （2% DMSO）; （3） positive control group （50 mg/L 5-Fluorouracil, 5-FU）; （4） low-dose group （LSC, final concentration of schisandrin B, 25 mg/L）, （5） moderate-dose group （MSC, final concentration of schisandrin B, 50 mg/L）; （6） highdose group （HSC, final concentration of schisandrin B, 100 mg/L）. Follow-up was done at 12-48 h. An MTT （Methylthiazolyldiphenyl-tetrazolium bromide） assay was used to examine the inhibitory effect of SOB on gastric cancer cells. The mitosis index was assessed using an inverted microscope. Flow cytometry was used to visualize the cell cycle. An RT-PCR （Reverse transcription-Polymerase chain reaction） -based assay was used to detect mRNA expression for cyclin D1 and glyceraldehyde-3-phosphate dehydrogenase （GAPDH）.RESULTS： The MTT assay showed that the number of living cells in the LSC, MSC and HSC groups was significantly smaller than that in the DMSO-treated group （P 〈 0.05） at 12-48 h. The inhibitory rate （IR） of the LSC group was 41.15% ± 3.86%, 59.24% ± 5.34% and 69.93% ± 7.81% at 12, 24 and 48 h, respectively. The IR of the MSC group was 42.82% ± 4.94%, 62.68% ± 7.58% and 71.79% ± 8.12% at 12, 24 and 48 h, respectively. The IR of the HSC group was 37.50% ± 3.21%, 40.34% ± 2.98% and 61.99% ± 4.88% at 12, 24 and 48 h, respectively. These results suggested that a moderate dosage had the most obvious inhibitory efficacy at 48 h. Compared to the DMSO group, the mitosis index of the LSC, MSC, HSC groups was greatly decreased （P 〈 0.05） at all time points. Any dose of SC-B suppressed mitosis within 12-48 h. Compared to the DMSO group, the percentage of cells in the G0/G1 phase of the MSC group was greatly increased, and that of the S ＋ G2M phase was greatly decreased, while the percentage of cell inhibition （PCI） in the MSC group was greatly increased （P 〈 0.05）. This suggested that SC-B could exclusively arrest cells in the G0/G1 phase. Cyclin D1 mRNA expression was lower in the MSC group than that in the DMSO group （P 〈 0.05）.CONCLUSION： SC-B can inhibit the proliferation and aberrant mitosis of human gastric cancer SCG-7901 cells /n v/tro, This inhibitory effect may be due to the down- regulation of cyclin D1 mRNA expression, which causes cell cycle arrest of gastric cancer cells.展开更多
AIM:To investigate the effects of schisandrin B (Sch B) on free fatty acid (FFA)-induced steatosis in L-02 cells.METHODS:Cellular steatosis was induced by incubating L-02 cells with a FFA mixture (oleate and palmitate...AIM:To investigate the effects of schisandrin B (Sch B) on free fatty acid (FFA)-induced steatosis in L-02 cells.METHODS:Cellular steatosis was induced by incubating L-02 cells with a FFA mixture (oleate and palmitate at the ratio of 2:1) for 24 h.Cytotoxicity and apoptosis were evaluated by 3-(4,5-dmethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay and Annexin V/propidium iodide staining,respectively.Cellular total lipid was determined using a photocolorimetric method after Nile red staining,and triglyceride content was measured using an enzymatic kit.To study the effects of Sch B on steatosis,L-02 cells were treated with Sch B (1-100 μmol/L) in the absence or presence of 1 mmol/L FFA for 24 h,and cellular total lipid and triglyceride levels were measured.To explore the mechanisms of action of Sch B in the steatotic L-02 cells,mRNA levels of several regulators of hepatic lipid metabolism including adipose differentiation related protein (ADRP),sterol regulatory element binding protein 1 (SREBP-1),peroxisome proliferator-activated receptor (PPAR)-α and PPAR-γ were measured by quantitative real-time polymerase chain reaction (PCR),and protein levels of ADRP and SREBP-1 were measured by immunoblotting.RESULTS:Treatment with 1 mmol/L FFA for 24 h induced intracellular lipid accumulation in L-02 cells comparable to that in human steatotic livers without causing apparent apoptosis and cytotoxicity.Sch B mitigated cellular total lipid and triglyceride accumulations in the steatotic L-02 cells in a dose-dependent manner.Quantitative real-time PCR and Western blot analyses revealed that treatment of L-02 cells with 100 μmol/L Sch B reverted the FFA-stimulated up-regulation of ADRP and SREBP-1.CONCLUSION:Sch B inhibits FFA-induced steatosis in L-02 cells by,at least in part,reversing the up-regulation of ADRP and SREBP-1.展开更多
Schisandrin B(Sch B)is a monomer with anti-cancer and anti-inflammatory effects,which are isolated from the plant Schisandra chinensis(Turcz)Baillon.We investigated the anti-gastric cancer(GC)effects of Sch B and its ...Schisandrin B(Sch B)is a monomer with anti-cancer and anti-inflammatory effects,which are isolated from the plant Schisandra chinensis(Turcz)Baillon.We investigated the anti-gastric cancer(GC)effects of Sch B and its underlying molecular mechanisms.The Cell Counting Kit-8 assay was used to determine the effects of Sch B on the viability of GC and normal cell lines.Hoechst/propidium iodide staining and flow cytometry were used to assess the apoptosis induction of Sch B.Western blotting was used to evaluate the effects of Sch B on downstream apoptotic proteins.The DCFH-DA fluorescent probe was used to assess the regulatory effects of Sch B on reactive oxygen species(ROS)levels and related signaling pathways in GC cells.The results showed that Sch B could regulate the phosphorylation level of mitogen-activated protein kinase(MAPK)by upregulating ROS accumulation in gastric cancer cells,and then reduce the expression of nuclear factor kappa B(NF-κB)and phosphorylated transcription 3(p-STAT3).In addition,Sch B downregulated the cell cycle proteins cyclin-dependent kinase 2/4/6 and cyclin D1/E,and arrested cells in the G0/G1 phase.Moreover,it also inhibited cell migration,which was reversed with Nacetylcysteine pretreatment.In summary,Sch B has killing effects on GC cells by upregulating the production of intracellular ROS and regulating the MAPK/STAT3/NF-κB signaling pathway,leading to the migration arrest and apoptosis of GC cells.展开更多
Schisandrin B (Sch B) is one of the active dibenzocyclooctadiene lignans found in the Schisandrae Fructus. Experimental studies have shown that Sch B possesses various pharmacological properties, including anti-cancer...Schisandrin B (Sch B) is one of the active dibenzocyclooctadiene lignans found in the Schisandrae Fructus. Experimental studies have shown that Sch B possesses various pharmacological properties, including anti-cancer, neuroprotective and nephroprotective activities. However, no detailed information on its biotransformation was reported in the literature. Here, we investigated the in vitro and in vivo metabolism of Sch B by using ultra-performance liquid chromatography coupled with tandem mass spectrometry. In vitro study detected and identified one oxygenated metabolite. Four metabolites were detected and identified from the in vivo study. The results indicated that the metabolism of Sch B mainly involved the demethylation of methoxy groups, the opening of five-member ring and the glucuronidation of metabolites in rats. The metabolites were identified for the first time by MS/MS analyses.展开更多
PC12 cell injury was induced using 20 μM amyloid β-protein 25-35 to establish a model of Alzheimer＇s disease. The cells were then treated with 5, 10, and 25 μM Schisandrin B. Methylthiazolyldiphenyl-tetrazolium br...PC12 cell injury was induced using 20 μM amyloid β-protein 25-35 to establish a model of Alzheimer＇s disease. The cells were then treated with 5, 10, and 25 μM Schisandrin B. Methylthiazolyldiphenyl-tetrazolium bromide assays and Hoechst 33342 staining results showed that with increasing Schisandrin B concentration, the survival rate of PC12 cells injured by amyloid β-protein 25-35 gradually increased and the rate of apoptosis gradually decreased. Reverse transcription-PCR, immunocytochemical staining and western blot results showed that with increasing Schisandrin B concentration, the mRNA and protein expression of vacuolar protein sorting 35 and amyloid precursor protein were gradually decreased. Vacuolar protein sorting 35 and amyloid precursor protein showed a consistent trend for change. These findings suggest that 5, 10, and 25 μM Schisandrin B antagonizes the cellular injury induced by amyloid β-protein 25-35 in a dose-dependent manner. This may be caused by decreasing the expression of vacuolar protein sorting 35 and amyloid precursor protein.展开更多
The determination method of Schisandrin A and Schisandrin B in Schisandra chinensis was improved with the high performance liquid chromagraphy （HPLC）. The sample was extracted exceedingly in the critical limit of CO...The determination method of Schisandrin A and Schisandrin B in Schisandra chinensis was improved with the high performance liquid chromagraphy （HPLC）. The sample was extracted exceedingly in the critical limit of CO2. The retention time of Schisandrin A and Schisandrin B was reduced, with methano/water （75 ： 25） as mobile phase. The wavelength for detection was 254 nm. The R^2 of standard curve was 0.9998 and the relative standard deviation was 2.31% and 3.17% with the recovery of 96.45% and 97.37%, respectively. The result shows that the rate of veracity of this method is higher and it proves that the determination method of Sehisandrin A and Schisandrin B in Schisandra chinensis is a feasible method.展开更多
Schisandrin A is a natural dibenzocyclooctene lignan with potent neuroprotective activity.However,the specific mechanisms or direct target proteins have not been clarified up to now.In this study,we designed and synth...Schisandrin A is a natural dibenzocyclooctene lignan with potent neuroprotective activity.However,the specific mechanisms or direct target proteins have not been clarified up to now.In this study,we designed and synthesized the probes of schisandrin A with photoreactive diazirine and clickable alkyne to identify its direct target in SH-SY5Y cells by employing activity-based protein profiling(ABPP)technique.Ykt6 was prominent among the 13 proteins obtained with high confidence and we confirmed Ykt6 as the direct target of schisandrin A by CETSA,IF,SPR and knockdown assay.Functionally,schisandrin A protected the cells against the injury induced by glutamate by regulating autophagy via Ykt6.This discovery may provide a novel therapeutic option for various neuronal cell damage-mediated diseases.展开更多
Schisandrae Fructus, containing schisandrin B (Sch B) as its main active component, is recognized in traditional Chinese medicine (TCM) for its Qi-invigorating properties in the five visceral organs. Our laboratory ha...Schisandrae Fructus, containing schisandrin B (Sch B) as its main active component, is recognized in traditional Chinese medicine (TCM) for its Qi-invigorating properties in the five visceral organs. Our laboratory has shown that the Qi-invigorating action of Chinese tonifying herbs is linked to increased mitochondrial ATP generation and an enhancement in mitochondrial glutathione redox status. To explore whether Sch B can exert Qi-invigorating actions across various tissues, we investigated the effects of Sch B treatment on mitochondrial ATP generation and glutathione redox status in multiple mouse tissues ex vivo. In line with TCM theory, which posits that Zheng Qi generation relies on the Qi function of the visceral organs, we also examined Sch B’s impact on natural killer cell activity and antigen-induced splenocyte proliferation, both serving as indirect measures of Zheng Qi. Our findings revealed that Sch B treatment consistently enhanced mitochondrial ATP generation and improved mitochondrial glutathione redox status in mouse tissues. This boost in mitochondrial function was associated with stimulated innate and adaptive immune responses, marked by increased natural killer cell activity and antigen-induced T/B cell proliferation, potentially through the increased generation of Zheng Qi.展开更多
Oligoasthenospermia is the primary cause of infertility.However,there are still enormous challenges in the screening of critical candidates and targets of oligoasthenospermia owing to its complex mechanism.In this stu...Oligoasthenospermia is the primary cause of infertility.However,there are still enormous challenges in the screening of critical candidates and targets of oligoasthenospermia owing to its complex mechanism.In this study,stem cell factor(SCF),c-kit,and transient receptor potential vanilloid 1(TRPV1)biosensors were successfully established and applied to studying apoptosis and autophagy mechanisms.Interestingly,the detection limit reached 2.787×10^(-15)g/L,and the quantitative limit reached 1.0×10^(-13)g/L.Furthermore,biosensors were used to investigate the interplay between autophagy and apoptosis.Schisandrin A is an excellent candidate to form a system with c-kit similar to SCF/c-kit with a detection constant(K_(D))of 5.701×10^(-11)mol/L,whereas it had no affinity for SCF.In addition,it also inhibited autophagy in oligoasthenospermia through antagonizing TRPV1 with a K_(D) of up to 4.181×10^(-10)mol/L.In addition,in vivo and in vitro experiments were highly consistent with the biosensor.In summary,high-potency schisandrin A and two potential targets were identified,through which schisandrin A could reverse the apoptosis caused by excessive autophagy during oligoasthenospermia.Our study provides promising insights into the discovery of effective compounds and potential targets via a well-established in vitro-in vivo strategy.展开更多
Objective To explore the effect and mechanism of schisandrin B(Sch B)in the treatment of cerebral ischemia in rats.Methods The cerebral ischemia models were induced by middle cerebral artery occlusion(MCAO)and reperfu...Objective To explore the effect and mechanism of schisandrin B(Sch B)in the treatment of cerebral ischemia in rats.Methods The cerebral ischemia models were induced by middle cerebral artery occlusion(MCAO)and reperfusion.Sprague-Dawley rats were divided into 6 groups using a random number table,including sham,MCAO,MCAO+Sch B(50 mg/kg),MCAO+Sch B(100 mg/kg),MCAO+Sch B(100 mg/kg)+LY294002,and MCAO+Sch B(100 mg/kg)+wortmannin groups.The effects of Sch B on pathological indicators,including neurological deficit scores,cerebral infarct volume,and brain edema,were subsequently studied.Tissue apoptosis was identified by terminal transferase-mediated dUTP nick end-labeling(TUNEL)staining.The protein expressions involved in apoptosis,inflammation response and oxidative stress were examined by immunofluorescent staining,biochemical analysis and Western blot analysis,respectively.The effect of Sch B on phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)signaling was also explored.Results Sch B treatment decreased neurological deficit scores,cerebral water content,and infarct volume in MCAO rats(P<0.05 or P<0.01).Neuronal nuclei and TUNEL staining indicated that Sch B also reduced apoptosis in brain tissues,as well as the Bax/Bcl-2 ratio and caspase-3 expression(P<0.01).Sch B regulated the production of myeloperoxidase,malondialdehyde,nitric oxide and superoxide dismutase,as well as the release of cytokine interleukin(IL)-1βand IL-18,in MCAO rats(P<0.05 or P<0.01).Sch B promoted the phosphorylation of PI3K and AKT.Blocking the PI3K/AKT signaling pathway with LY294002 or wortmannin reduced the protective effect of Sch B against cerebral ischemia(P<0.05 or P<0.01).Conclusions Sch B reduced apoptosis,inflammatory response,and oxidative stress of MCAO rats by modulating the PI3K/AKT pathway.Sch B had a potential for treating cerebral ischemia.展开更多
Objective: To determine whether Schisandrin B(Sch B) attenuates early brain injury(EBI) in rats with subarachnoid hemorrhage(SAH). Methods: Sprague-Dawley rats were divided into sham(sham operation), SAH, SAH+vehicle,...Objective: To determine whether Schisandrin B(Sch B) attenuates early brain injury(EBI) in rats with subarachnoid hemorrhage(SAH). Methods: Sprague-Dawley rats were divided into sham(sham operation), SAH, SAH+vehicle, and SAH+Sch B groups using a random number table. Rats underwent SAH by endovascular perforation and received Sch B(100 mg/kg) or normal saline after 2 and 12 h of SAH. SAH grading, neurological scores, brain water content, Evan’s blue extravasation, and terminal transferase-mediated dUTP nick end-labeling(TUNEL) staining were carried out 24 h after SAH. Immunofluorescent staining was performed to detect the expressions of ionized calcium binding adapter molecule 1(Iba-1) and myeloperoxidase(MPO) in the rat brain, while the expressions of B-cell lymphoma 2(Bcl-2), Bax, Caspase-3, nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3(NLRP3), apoptosis-associated specklike protein containing the caspase-1 activator domain(ASC), Caspase-1, interleukin(IL)-1β, and IL-18 in the rat brains were detected by Western blot. Results: Compared with the SAH group, Sch B significantly improved the neurological function, reduced brain water content, Evan’s blue content, and apoptotic cells number in the brain of rats(P<0.05 or P<0.01). Moreover, Sch B decreased SAH-induced expressions of Iba-1 and MPO(P<0.01). SAH caused the elevated expressions of Bax, Caspase-3, NLRP3, ASC, Caspase-1, IL-1β, and IL-18 in the rat brain(P<0.01), all of which were inhibited by Sch B(P<0.01). In addition, Sch B increased the Bcl-2 expression(P<0.01). Conclusion: Sch B attenuated SAH-induced EBI, which might be associated with the inhibition of neuroinflammation, neuronal apoptosis, and the NLRP3 inflammatory signaling pathway.展开更多
Abstract Objective To determine the effect of dimethyl 4, 4' dimethoxy 5, 6, 5', 6 dimethylene dioxybiphenyl 2, 2' dicarboxylate (HpPro) on patients with acute and chronic liver diseases. Methods...Abstract Objective To determine the effect of dimethyl 4, 4' dimethoxy 5, 6, 5', 6 dimethylene dioxybiphenyl 2, 2' dicarboxylate (HpPro) on patients with acute and chronic liver diseases. Methods An open trial and a prospective randomized and controlled study were performed. The open trial consisted of 56 cases (16 cases of acute hepatitis, 20 cases of chronic hepatitis, 14 cases of liver cirrhosis and 6 cases of fatty liver). Controlled study consisted of 20 cases of Child A chronic hepatitis which were randomly treated with either HpPro or a mixture of known drugs which used as a liver protective agent in Indonesia as control for one week. The patients were then crossed over those two drugs in the next week. Results In the open trial, after 4 weeks' treatment with HpPro 7.5 mg orally three times daily, acute hepatitis, chronic hepatitis and fatty liver cases showed rapid decrease of SGOT and SGPT. In the liver cirrhosis cases, SGOT and SGPT were decreased slowly. In the controlled trial, nine patients received HpPro 7.5 mg three times daily orally and eleven were treated with a mixture of known drugs as the controls. After one week treatment, HpPro group clinically showed significant decrease of SGPT and SGOT levels compared to control group (P=0.035). At the second week, HpPro group showed significant decrease of SGOT compared to control group (P=0.038) but the decrease of SGPT was not significant (P=0.096). Conclusion Treatment with HpPro is effective to reduce liver impairment in acute and chronic liver diseases on Indonesian patients. No side effect of HpPro was observed.展开更多
基金supported by the National Research Foundation of Korea grant funded by the Korea government(RS-2023-00213236).
文摘Objective:To investigate the synergistic effects of auranofin and schisandrin A(SA)on cell proliferation inhibition and apoptosis induction in human hepatocellular carcinoma Hep3B cells.Methods:Cell viability was assessed using MTT to determine the synergistic effects of auranofin and SA.Three-dimensional(3D)culture models were used to evaluate the effects on spheroid structure and size.Apoptosis was analyzed by flow cytometry for sub-G1 populations,annexin V staining,and Western blotting for apoptotic markers.Reactive oxygen species(ROS)production was measured using DCF-DA staining.Results:Our results showed that combined treatment with auranofin and SA led to a significant reduction in cell viability compared with either compound alone,with isobologram analysis confirming their synergistic interactions.Under 3D culture conditions,auranofin and SA disrupted the compact structure of spheroids,leading to a loosened and disorganized morphology at the periphery,which appeared as an increase in spheroid size.Moreover,the induction of apoptosis by auranofin and SA was evidenced by elevated sub-G1 phase populations,increased annexin V-positive cells,and upregulation of apoptotic markers such as cleaved poly(ADPribose)polymerase 1 and cleaved caspase-3.Notably,auranofin combined with SA markedly enhanced ROS production,which was mitigated by the ROS scavenger N-acetylcysteine.Additionally,the phosphoinositide 3-kinase(PI3K)/Akt signaling pathway was downregulated in response to auranofin and SA treatment,and further apoptotic effects were observed following PI3K inhibition with LY294002.Conclusions:Auranofin combined with SA promotes apoptosis of hepatocellular carcinoma via ROS generation and inhibition of the PI3K/Akt pathway.
基金supported by the National Natural Science Foundation of China(No.81274004)National Key Technologies R&D Program of China(No.2008BAI51B03)+1 种基金2011’Program for Excellent Scientific and Technological Innovation Team of Jiangsu Higher Education,a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions,the Project Program of the State Key Laboratory of Natural Medicines,China Pharmaceutical University(No.JKGZ201107)the Graduate Student Scientific Research Innovation Plan of Jiangsu Higher Education Institutions(No.CXZZ11_0795)
文摘AIM： To illuminate the molecular targets for schisandrin against cerebrovascular disease based on the combined methods of network pharmacology prediction and experimental verification. METHOD： A protein database was established through constructing the drug-protein network from literature mining data. The protein-protein network was built through an in-depth exploration of the relationships between the proteins. The computational platform was implemented to predict and extract the sensitive sub-network with significant P-values from the protein-protein network. Then the key targets and pathways were identified from the sensitive sub-network. The most related targets and pathways were also confirmed in hydrogen peroxide （H202）-induced PC 12 cells by Western blotting. RESULTS： Twelve differentially expressed proteins （gene names： NFKB1, RELA, TNFSF10, MAPK1, CHUK, CASP8, PIGS2, MAPK 14, CREBI, IFNG, APR and BCL2） were confirmed as the central nodes of the interaction network （45 nodes, 93 edges）. The NF-KB signaling pathway was suggested as the most related pathway of schisandrin for cerebrovascular disease. Furthermore, schisandrin was found to suppress the expression and phosphorylation of 1KKct, as well as p50 and p65 induced by H2O2 in PC12 cells by Western blotting. CONCLUSION： The computational platform that integrates literature mining data, protein-protein interactions, sensitive sub-network, and pathway results in identification of the NF-arB signaling pathway as the key targets and pathways for schisandrin.
基金Supported by The Project of Heilongjiang Natural Science Foundation No. TD 2005-01Department of Health, Heilongjiang Province No. 2006-391
文摘AIM： To investigate the inhibitory effect and possible mechanism of action of schisandrin B in SC-B on gastric cancer cells in vitro.METHODS： SC-B consisted of schisandrin B, aloeemodin, and Astragalus polysaccharides. Exponentially growing human gastric cancer SGO7901 cells were divided into six treatment groups： （1） control group （RPMI 1640 medium）; （2） negative control group （2% DMSO）; （3） positive control group （50 mg/L 5-Fluorouracil, 5-FU）; （4） low-dose group （LSC, final concentration of schisandrin B, 25 mg/L）, （5） moderate-dose group （MSC, final concentration of schisandrin B, 50 mg/L）; （6） highdose group （HSC, final concentration of schisandrin B, 100 mg/L）. Follow-up was done at 12-48 h. An MTT （Methylthiazolyldiphenyl-tetrazolium bromide） assay was used to examine the inhibitory effect of SOB on gastric cancer cells. The mitosis index was assessed using an inverted microscope. Flow cytometry was used to visualize the cell cycle. An RT-PCR （Reverse transcription-Polymerase chain reaction） -based assay was used to detect mRNA expression for cyclin D1 and glyceraldehyde-3-phosphate dehydrogenase （GAPDH）.RESULTS： The MTT assay showed that the number of living cells in the LSC, MSC and HSC groups was significantly smaller than that in the DMSO-treated group （P 〈 0.05） at 12-48 h. The inhibitory rate （IR） of the LSC group was 41.15% ± 3.86%, 59.24% ± 5.34% and 69.93% ± 7.81% at 12, 24 and 48 h, respectively. The IR of the MSC group was 42.82% ± 4.94%, 62.68% ± 7.58% and 71.79% ± 8.12% at 12, 24 and 48 h, respectively. The IR of the HSC group was 37.50% ± 3.21%, 40.34% ± 2.98% and 61.99% ± 4.88% at 12, 24 and 48 h, respectively. These results suggested that a moderate dosage had the most obvious inhibitory efficacy at 48 h. Compared to the DMSO group, the mitosis index of the LSC, MSC, HSC groups was greatly decreased （P 〈 0.05） at all time points. Any dose of SC-B suppressed mitosis within 12-48 h. Compared to the DMSO group, the percentage of cells in the G0/G1 phase of the MSC group was greatly increased, and that of the S ＋ G2M phase was greatly decreased, while the percentage of cell inhibition （PCI） in the MSC group was greatly increased （P 〈 0.05）. This suggested that SC-B could exclusively arrest cells in the G0/G1 phase. Cyclin D1 mRNA expression was lower in the MSC group than that in the DMSO group （P 〈 0.05）.CONCLUSION： SC-B can inhibit the proliferation and aberrant mitosis of human gastric cancer SCG-7901 cells /n v/tro, This inhibitory effect may be due to the down- regulation of cyclin D1 mRNA expression, which causes cell cycle arrest of gastric cancer cells.
基金Supported by The Hong Kong Baptist University,No.FRG 08-09/II-30
文摘AIM:To investigate the effects of schisandrin B (Sch B) on free fatty acid (FFA)-induced steatosis in L-02 cells.METHODS:Cellular steatosis was induced by incubating L-02 cells with a FFA mixture (oleate and palmitate at the ratio of 2:1) for 24 h.Cytotoxicity and apoptosis were evaluated by 3-(4,5-dmethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay and Annexin V/propidium iodide staining,respectively.Cellular total lipid was determined using a photocolorimetric method after Nile red staining,and triglyceride content was measured using an enzymatic kit.To study the effects of Sch B on steatosis,L-02 cells were treated with Sch B (1-100 μmol/L) in the absence or presence of 1 mmol/L FFA for 24 h,and cellular total lipid and triglyceride levels were measured.To explore the mechanisms of action of Sch B in the steatotic L-02 cells,mRNA levels of several regulators of hepatic lipid metabolism including adipose differentiation related protein (ADRP),sterol regulatory element binding protein 1 (SREBP-1),peroxisome proliferator-activated receptor (PPAR)-α and PPAR-γ were measured by quantitative real-time polymerase chain reaction (PCR),and protein levels of ADRP and SREBP-1 were measured by immunoblotting.RESULTS:Treatment with 1 mmol/L FFA for 24 h induced intracellular lipid accumulation in L-02 cells comparable to that in human steatotic livers without causing apparent apoptosis and cytotoxicity.Sch B mitigated cellular total lipid and triglyceride accumulations in the steatotic L-02 cells in a dose-dependent manner.Quantitative real-time PCR and Western blot analyses revealed that treatment of L-02 cells with 100 μmol/L Sch B reverted the FFA-stimulated up-regulation of ADRP and SREBP-1.CONCLUSION:Sch B inhibits FFA-induced steatosis in L-02 cells by,at least in part,reversing the up-regulation of ADRP and SREBP-1.
基金This work was supported by the National Natural Science Foundation of China[Grant No.82060118]the Research Program of Science and Technology at Universities of Inner Mongolia Autonomous Region[Grant No.NJZY20203]+3 种基金the Program for Young Talents of Chifeng University[Grant No.CFXYYT2202]the Central Government Supports Local College Reform and Development Fund Talent Training Projects[Grant No.2020GSP16]the Heilongjiang Touyan Innovation Team Program[Grant No.2019HTY078]the Project for Heilongjiang Bayi Agricultural University[Grant No.XDB202012].
文摘Schisandrin B(Sch B)is a monomer with anti-cancer and anti-inflammatory effects,which are isolated from the plant Schisandra chinensis(Turcz)Baillon.We investigated the anti-gastric cancer(GC)effects of Sch B and its underlying molecular mechanisms.The Cell Counting Kit-8 assay was used to determine the effects of Sch B on the viability of GC and normal cell lines.Hoechst/propidium iodide staining and flow cytometry were used to assess the apoptosis induction of Sch B.Western blotting was used to evaluate the effects of Sch B on downstream apoptotic proteins.The DCFH-DA fluorescent probe was used to assess the regulatory effects of Sch B on reactive oxygen species(ROS)levels and related signaling pathways in GC cells.The results showed that Sch B could regulate the phosphorylation level of mitogen-activated protein kinase(MAPK)by upregulating ROS accumulation in gastric cancer cells,and then reduce the expression of nuclear factor kappa B(NF-κB)and phosphorylated transcription 3(p-STAT3).In addition,Sch B downregulated the cell cycle proteins cyclin-dependent kinase 2/4/6 and cyclin D1/E,and arrested cells in the G0/G1 phase.Moreover,it also inhibited cell migration,which was reversed with Nacetylcysteine pretreatment.In summary,Sch B has killing effects on GC cells by upregulating the production of intracellular ROS and regulating the MAPK/STAT3/NF-κB signaling pathway,leading to the migration arrest and apoptosis of GC cells.
文摘Schisandrin B (Sch B) is one of the active dibenzocyclooctadiene lignans found in the Schisandrae Fructus. Experimental studies have shown that Sch B possesses various pharmacological properties, including anti-cancer, neuroprotective and nephroprotective activities. However, no detailed information on its biotransformation was reported in the literature. Here, we investigated the in vitro and in vivo metabolism of Sch B by using ultra-performance liquid chromatography coupled with tandem mass spectrometry. In vitro study detected and identified one oxygenated metabolite. Four metabolites were detected and identified from the in vivo study. The results indicated that the metabolism of Sch B mainly involved the demethylation of methoxy groups, the opening of five-member ring and the glucuronidation of metabolites in rats. The metabolites were identified for the first time by MS/MS analyses.
基金supported by the National 985 Project "linguistic science technology and the construction of interdisciplinary innovation platform in current society",No.985yk002the National 985 Project "cognitive and neural information science platform",No.904273258
文摘PC12 cell injury was induced using 20 μM amyloid β-protein 25-35 to establish a model of Alzheimer＇s disease. The cells were then treated with 5, 10, and 25 μM Schisandrin B. Methylthiazolyldiphenyl-tetrazolium bromide assays and Hoechst 33342 staining results showed that with increasing Schisandrin B concentration, the survival rate of PC12 cells injured by amyloid β-protein 25-35 gradually increased and the rate of apoptosis gradually decreased. Reverse transcription-PCR, immunocytochemical staining and western blot results showed that with increasing Schisandrin B concentration, the mRNA and protein expression of vacuolar protein sorting 35 and amyloid precursor protein were gradually decreased. Vacuolar protein sorting 35 and amyloid precursor protein showed a consistent trend for change. These findings suggest that 5, 10, and 25 μM Schisandrin B antagonizes the cellular injury induced by amyloid β-protein 25-35 in a dose-dependent manner. This may be caused by decreasing the expression of vacuolar protein sorting 35 and amyloid precursor protein.
基金Supported by National Basic Research Program of China(2004CB11750-5)
文摘The determination method of Schisandrin A and Schisandrin B in Schisandra chinensis was improved with the high performance liquid chromagraphy （HPLC）. The sample was extracted exceedingly in the critical limit of CO2. The retention time of Schisandrin A and Schisandrin B was reduced, with methano/water （75 ： 25） as mobile phase. The wavelength for detection was 254 nm. The R^2 of standard curve was 0.9998 and the relative standard deviation was 2.31% and 3.17% with the recovery of 96.45% and 97.37%, respectively. The result shows that the rate of veracity of this method is higher and it proves that the determination method of Sehisandrin A and Schisandrin B in Schisandra chinensis is a feasible method.
基金supported by the CAMS Innovation Fund for Medical Sciences(CIFMS,Nos.2021-I2M-1-069,2022-12M-2-002)Beijing Municipal Natural Science Foundation,China(No.7222259)。
文摘Schisandrin A is a natural dibenzocyclooctene lignan with potent neuroprotective activity.However,the specific mechanisms or direct target proteins have not been clarified up to now.In this study,we designed and synthesized the probes of schisandrin A with photoreactive diazirine and clickable alkyne to identify its direct target in SH-SY5Y cells by employing activity-based protein profiling(ABPP)technique.Ykt6 was prominent among the 13 proteins obtained with high confidence and we confirmed Ykt6 as the direct target of schisandrin A by CETSA,IF,SPR and knockdown assay.Functionally,schisandrin A protected the cells against the injury induced by glutamate by regulating autophagy via Ykt6.This discovery may provide a novel therapeutic option for various neuronal cell damage-mediated diseases.
文摘Schisandrae Fructus, containing schisandrin B (Sch B) as its main active component, is recognized in traditional Chinese medicine (TCM) for its Qi-invigorating properties in the five visceral organs. Our laboratory has shown that the Qi-invigorating action of Chinese tonifying herbs is linked to increased mitochondrial ATP generation and an enhancement in mitochondrial glutathione redox status. To explore whether Sch B can exert Qi-invigorating actions across various tissues, we investigated the effects of Sch B treatment on mitochondrial ATP generation and glutathione redox status in multiple mouse tissues ex vivo. In line with TCM theory, which posits that Zheng Qi generation relies on the Qi function of the visceral organs, we also examined Sch B’s impact on natural killer cell activity and antigen-induced splenocyte proliferation, both serving as indirect measures of Zheng Qi. Our findings revealed that Sch B treatment consistently enhanced mitochondrial ATP generation and improved mitochondrial glutathione redox status in mouse tissues. This boost in mitochondrial function was associated with stimulated innate and adaptive immune responses, marked by increased natural killer cell activity and antigen-induced T/B cell proliferation, potentially through the increased generation of Zheng Qi.
基金co-supported by National Outstanding Youth Science Fund Project of National Natural Science Foundation of China(No.82022073)National Natural Science Foundation of China(No.82174389)+1 种基金Natural Science Foundation of Beijing Municipality(No.7202115,China)Postdoctoral Research Foundation of China(No.2021M690474)。
文摘Oligoasthenospermia is the primary cause of infertility.However,there are still enormous challenges in the screening of critical candidates and targets of oligoasthenospermia owing to its complex mechanism.In this study,stem cell factor(SCF),c-kit,and transient receptor potential vanilloid 1(TRPV1)biosensors were successfully established and applied to studying apoptosis and autophagy mechanisms.Interestingly,the detection limit reached 2.787×10^(-15)g/L,and the quantitative limit reached 1.0×10^(-13)g/L.Furthermore,biosensors were used to investigate the interplay between autophagy and apoptosis.Schisandrin A is an excellent candidate to form a system with c-kit similar to SCF/c-kit with a detection constant(K_(D))of 5.701×10^(-11)mol/L,whereas it had no affinity for SCF.In addition,it also inhibited autophagy in oligoasthenospermia through antagonizing TRPV1 with a K_(D) of up to 4.181×10^(-10)mol/L.In addition,in vivo and in vitro experiments were highly consistent with the biosensor.In summary,high-potency schisandrin A and two potential targets were identified,through which schisandrin A could reverse the apoptosis caused by excessive autophagy during oligoasthenospermia.Our study provides promising insights into the discovery of effective compounds and potential targets via a well-established in vitro-in vivo strategy.
基金the Natural Science Foundation of Fujian Province of China(No.2022J01245)the Quanzhou City Science&Technology Program of China(No.2020N020s)the Excellent Young Scholars Cultivation Project of Fujian Medical University Union Hospital(No.2022XH040)。
文摘Objective To explore the effect and mechanism of schisandrin B(Sch B)in the treatment of cerebral ischemia in rats.Methods The cerebral ischemia models were induced by middle cerebral artery occlusion(MCAO)and reperfusion.Sprague-Dawley rats were divided into 6 groups using a random number table,including sham,MCAO,MCAO+Sch B(50 mg/kg),MCAO+Sch B(100 mg/kg),MCAO+Sch B(100 mg/kg)+LY294002,and MCAO+Sch B(100 mg/kg)+wortmannin groups.The effects of Sch B on pathological indicators,including neurological deficit scores,cerebral infarct volume,and brain edema,were subsequently studied.Tissue apoptosis was identified by terminal transferase-mediated dUTP nick end-labeling(TUNEL)staining.The protein expressions involved in apoptosis,inflammation response and oxidative stress were examined by immunofluorescent staining,biochemical analysis and Western blot analysis,respectively.The effect of Sch B on phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)signaling was also explored.Results Sch B treatment decreased neurological deficit scores,cerebral water content,and infarct volume in MCAO rats(P<0.05 or P<0.01).Neuronal nuclei and TUNEL staining indicated that Sch B also reduced apoptosis in brain tissues,as well as the Bax/Bcl-2 ratio and caspase-3 expression(P<0.01).Sch B regulated the production of myeloperoxidase,malondialdehyde,nitric oxide and superoxide dismutase,as well as the release of cytokine interleukin(IL)-1βand IL-18,in MCAO rats(P<0.05 or P<0.01).Sch B promoted the phosphorylation of PI3K and AKT.Blocking the PI3K/AKT signaling pathway with LY294002 or wortmannin reduced the protective effect of Sch B against cerebral ischemia(P<0.05 or P<0.01).Conclusions Sch B reduced apoptosis,inflammatory response,and oxidative stress of MCAO rats by modulating the PI3K/AKT pathway.Sch B had a potential for treating cerebral ischemia.
基金Supported by the Natural Science Foundation of Fujian Province of China (No. 2020J011016)the Joint Funds for the Innovation of Science and Technology of Fujian Province (No. 2018Y9004)the Startup Fund for Scientific Research of Fujian Medical University (No. 2019QH1055)。
文摘Objective: To determine whether Schisandrin B(Sch B) attenuates early brain injury(EBI) in rats with subarachnoid hemorrhage(SAH). Methods: Sprague-Dawley rats were divided into sham(sham operation), SAH, SAH+vehicle, and SAH+Sch B groups using a random number table. Rats underwent SAH by endovascular perforation and received Sch B(100 mg/kg) or normal saline after 2 and 12 h of SAH. SAH grading, neurological scores, brain water content, Evan’s blue extravasation, and terminal transferase-mediated dUTP nick end-labeling(TUNEL) staining were carried out 24 h after SAH. Immunofluorescent staining was performed to detect the expressions of ionized calcium binding adapter molecule 1(Iba-1) and myeloperoxidase(MPO) in the rat brain, while the expressions of B-cell lymphoma 2(Bcl-2), Bax, Caspase-3, nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3(NLRP3), apoptosis-associated specklike protein containing the caspase-1 activator domain(ASC), Caspase-1, interleukin(IL)-1β, and IL-18 in the rat brains were detected by Western blot. Results: Compared with the SAH group, Sch B significantly improved the neurological function, reduced brain water content, Evan’s blue content, and apoptotic cells number in the brain of rats(P<0.05 or P<0.01). Moreover, Sch B decreased SAH-induced expressions of Iba-1 and MPO(P<0.01). SAH caused the elevated expressions of Bax, Caspase-3, NLRP3, ASC, Caspase-1, IL-1β, and IL-18 in the rat brain(P<0.01), all of which were inhibited by Sch B(P<0.01). In addition, Sch B increased the Bcl-2 expression(P<0.01). Conclusion: Sch B attenuated SAH-induced EBI, which might be associated with the inhibition of neuroinflammation, neuronal apoptosis, and the NLRP3 inflammatory signaling pathway.
文摘Abstract Objective To determine the effect of dimethyl 4, 4' dimethoxy 5, 6, 5', 6 dimethylene dioxybiphenyl 2, 2' dicarboxylate (HpPro) on patients with acute and chronic liver diseases. Methods An open trial and a prospective randomized and controlled study were performed. The open trial consisted of 56 cases (16 cases of acute hepatitis, 20 cases of chronic hepatitis, 14 cases of liver cirrhosis and 6 cases of fatty liver). Controlled study consisted of 20 cases of Child A chronic hepatitis which were randomly treated with either HpPro or a mixture of known drugs which used as a liver protective agent in Indonesia as control for one week. The patients were then crossed over those two drugs in the next week. Results In the open trial, after 4 weeks' treatment with HpPro 7.5 mg orally three times daily, acute hepatitis, chronic hepatitis and fatty liver cases showed rapid decrease of SGOT and SGPT. In the liver cirrhosis cases, SGOT and SGPT were decreased slowly. In the controlled trial, nine patients received HpPro 7.5 mg three times daily orally and eleven were treated with a mixture of known drugs as the controls. After one week treatment, HpPro group clinically showed significant decrease of SGPT and SGOT levels compared to control group (P=0.035). At the second week, HpPro group showed significant decrease of SGOT compared to control group (P=0.038) but the decrease of SGPT was not significant (P=0.096). Conclusion Treatment with HpPro is effective to reduce liver impairment in acute and chronic liver diseases on Indonesian patients. No side effect of HpPro was observed.
