Objective This study aimed to explore the mechanism of obtaining yang from yin in Yougui pill against aging based on network pharmacology and molecular docking technology.Methods The active components and targets of Y...Objective This study aimed to explore the mechanism of obtaining yang from yin in Yougui pill against aging based on network pharmacology and molecular docking technology.Methods The active components and targets of Yougui Pill were obtained by searching the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)database and the Bioinformatics Analysis Tool for Molecular mechANism of Traditional Chinese Medicine(BATMEN-TCM)database,and kidney deficiency syndrome-related targets were obtained in the Symptom Mapping(SymMap)Database,a traditional Chinese medicine(TCM)syndrome correlation database.The protein–protein interaction(PPI)network was constructed by using the STRING11.5 database.Then,we used CytoScape3.9.0 software to construct the network of TCM–active components–potential targets,and the core TCM components and core targets of Yougui Pill for the treatment of kidney deficiency were obtained.The function analysis of Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathway enrichment analysis were performed in the Database for Annotation,Visualization,and Integrated Discovery(DAVID).Finally,preliminary verification was performed with the help of molecular docking technology.Results A total of 147 active components of 9 drugs of Yougui Pill(Fuzi[Aconiti Lateralis Radix Praeparata],Shudihaung[Rehmanniae Radix Praeparata],Gouqi[Lycii Fructus],Shanyao[Rhizoma Dioscoreae],Shanzhuyu[Corni Fructus],Tusizi[Cuscutae Semen],Danggui[Angelicae Sinensis Radix],Duzhong[Eucommiae Cortex],Lujiaojiao[Cervi Cornus Colla])were obtained,corresponding to 233 targets.A total of 2,235targets related to kidney deficiency syndrome and 43 potential therapeutic targets were obtained after the intersection.The core TCM components mainly included quercetin,kaempferol,diosgenin,β-carotene,etc.The core targets involved Trp53(Tp53),Akt1,Pparg,Nr3c1,App,Casp8,Mapk1,Cav1,and Ctnnb1.A total of 27biological processes,10 cellular components,and 11 molecular functions were obtained by gene function enrichment analysis,mainly related to the regulation of gene expression,cell apoptosis and proliferation,and the response to estrogen.A total of 51 KEGG signaling pathways,mainly involving a variety of cancer pathways,apoptosis pathways,longevity regulation pathways,etc.Conclusion Yougui Pill can play a role in preventing and treating kidney deficiency syndrome through multiple targets and pathways.展开更多
OBJECTIVE:To examine,dissect,and understand the molecular mechanisms and combinatorial effects of Zuogui(左归丸,ZGP)and Yougui pills(右归丸,YGP)in 4-vinyl cyclohexene diepoxide(4-VCD)-induced Perimenopausal syndrome(P...OBJECTIVE:To examine,dissect,and understand the molecular mechanisms and combinatorial effects of Zuogui(左归丸,ZGP)and Yougui pills(右归丸,YGP)in 4-vinyl cyclohexene diepoxide(4-VCD)-induced Perimenopausal syndrome(PMS).METHODS:Using the 4-VCD-induced PMS mouse model,uterine and ovary index were measured,and serum sex steroidal hormone levels were evaluated after treatment with ZGP,YGP,ZGP+YGP,estradiol valerate(EV),and Gengnian An(更年安,GNA).Histopathological examinations,ingredient-target network predictions,western blotting,and real-time quantitative polymerase chain reaction(RT-q PCR)analyses were performed to ascertain the possible pharmacological effects and molecular mechanisms of ZYP and YGP.RESULTS:Treatment with ZGP and YGP remarkably improves estrous cyclicity and prevents pathological damage to the uterus.Also,altered sex hormones,including AMH,E2,FSH,LH,P,and T,were restored to normal levels after ZGP and YGP administration.Ingredient-target network analysis showed that the 5 ingredients common to the ZGP and YGP formula modulate 53 targets shared with PMS.Pathwayenrichment analysis further predicted that ZGY and YGP likely regulate of apoptosis and other essential pathways during PMS.In-vivo studies showed that ZGP and YGP suppress PMS via modulating apoptosis through decreasing Caspase-3 and Bcl-2-associated X(Bax)levels and increasing B-cell lymphoma-2(Bcl-2)/Bax and Bcl-2 levels.Importantly,ZGP+YGP treatment modulation effects were somewhat or significantly better compared to ZGP or YGP alone treatment.CONCLUSION:ZGP and YGP represent novel anti-PMS agents whose effects involve restoring altered hormonal levels,protecting the uterus,and regulating apoptosis.展开更多
BACKGROUND Yougui pills have long been used to treat hypothyroidism,usually in combination with levothyroxine sodium in clinical treatment,while their clinical efficacy and safety are still controversial when compared...BACKGROUND Yougui pills have long been used to treat hypothyroidism,usually in combination with levothyroxine sodium in clinical treatment,while their clinical efficacy and safety are still controversial when compared to levothyroxine treatment alone.AIM To explore the clinical efficacy and safety of Yougui pills combined with levothyroxine sodium in the treatment of hypothyroidism.METHODS This meta-analysis was performed in accordance with the PRISMA guidelines.Randomized controlled trials on Yougui pills in the treatment of hypothyroidism published from 2008 to May 2021 were searched in a total of 8 databases(4 databases in Chinese and 4 databases in English).The quality of the included studies was evaluated according to the Cochrane risk assessment tool.Weighted mean difference(WMD)was used for continuous variables,and relative risk(RR)was used for binary variables.Data were extracted,and the meta-analysis was conducted with the statistical software of Stata15.0 and RevMan5.0.RESULTS A total of 140 articles were retrieved,and 9 of them were finally included,with a total sample size of 936 cases.The main meta-analysis results are as follows:(1)The group of Yougui pills combined with levothyroxine sodium had a significantly higher overall response rate than the group of levothyroxine sodium(RR=1.20,95%CI 1.12,1.28,P<0.00001);(2)Yougui pills combined with levothyroxine sodium achieved significantly better efficacy than levothyroxine sodium alone in alleviating adverse symptoms[standard mean difference(SMD)=-1.10,95%CI:-1.37,-0.84,P<0.00001];(3)The level of thyrotropin stimulating hormone in the group of Yougui pills combined with levothyroxine sodium was significantly lower than in the control group of levothyroxine sodium(WMD=-1.38,95%CI:-2.10,-0.67,P=0.00001);(4)The level of free triiodothyronine in the group of Yougui pills combined with levothyroxine sodium was higher than that in the control group of levothyroxine sodium(WMD=0.41,95%CI:0.03,0.79,P=0.03);(5)The level of free thyroxine in the group of Yougui pills combined with levothyroxine sodium was significantly higher than that in the control group of levothyroxine sodium(SMD=0.83,95%CI:0.44,1.22,P≤0.0001);and(6)The adverse reactions in the group of Yougui pills combined with levothyroxine sodium were significantly less than those in the control group of levothyroxine sodium(RR=0.33,95%CI:0.20,-0.53,P<0.00001).CONCLUSION In the treatment of hypothyroidism,the combination of Yougui pills with levothyroxine sodium may be better than levothyroxine sodium treatment alone.展开更多
Long-term glucocorticoid use may result in sustained suppression of one or more secreted components from the hypothalamo-pituitary-adrenal axis, and often results in apoptosis. Yougui Pill（YGP）, a 10-component tradi...Long-term glucocorticoid use may result in sustained suppression of one or more secreted components from the hypothalamo-pituitary-adrenal axis, and often results in apoptosis. Yougui Pill（YGP）, a 10-component traditional Chinese herbal medicine, has been shown to be clinically effective for glucocorticoid-induced suppression of the hypothalamo-pituitary-adrenal axis. However, the pharmacological and molecular mechanisms remain unclear. We hypothesized that YGP would exert an anti-apoptosis effect on dexamethasone-treated anterior pituitary cells. In vivo experiments showed that YGP significantly reduced the number of apoptotic cells, down-regulated m RNA expression of cytochrome c, caspase-3, and caspase-9, and up-regulated m RNA expression of Bcl-2. These findings suggest that YGP reduced glucocorticoid-induced apoptosis in rat anterior pituitary cells by regulating the mitochondria-mediated apoptosis pathway.展开更多
Objective: Bone marrow mesenchymal stem cells (BMSCs) have the potential to differentiate into chondrocytes, it is the ideal seed cells for treating cartilage injury. The purpose of this study was to investigate the i...Objective: Bone marrow mesenchymal stem cells (BMSCs) have the potential to differentiate into chondrocytes, it is the ideal seed cells for treating cartilage injury. The purpose of this study was to investigate the inducible factors that stimulate the differentiation of bone marrow mesenchymal stem cells into chondrocytes. Methods: Rat BMSCs were isolated and subcultured, and then inoculated on PLGA scaffolds after successfully identificating of BMSCs. They were divided into 5 groups. Blank serum group, TGF-β1 + Yougui Yin drug-containing serum induction group, TGF-β1 + mechanical stimulation induction group, Yougui Yin drug-containing serum + mechanical stimulation induction group, TGF-β1 + Yougui Yin drug-containing serum + mechanical stimulation induction group, 4 weeks later, the adhesion status of cells and scaffolds was observed by Scanning Electron Microscopy, and then chondrocyte induction was performed on different treatment groups. After induction, the expression of Aggrecan and Collagen II was detected by immunofluorescence, and the mRNA expression of Aggrecan and Collagen II was detected by qPCR. Results: BMSCs were identified as rat bone marrow mesenchymal stem cells by immunofluorescence CD34 and CD44 cells. Scanning Electron Microscopy observed that BMSCs grew well on PLGA. Compared with PLGA group, the scaffold space of BMSCS-PLGA composite was significantly reduced, and the growth of cell adhesion scaffold was significantly increased. Compared with the blank serum group, the expression of Aggrecan in TGF-β1 + Yougui Yin drug-containing serum group and Yougui Yin drug-containing serum + mechanical stimulation group was significantly increased. The expression of Collagen II in TGF-β1 + Yougui Yin medicated serum group, Yougui Yin medicated serum + mechanical stimulation group, TGF-β1 + mechanical stimulation group and TGF-β1 + Yougui Yin medicated serum + mechanical stimulation group were significantly increased;PCR results showed that compared with blank serum group, the expression of Aggrecan in other groups was significantly increased, except for Collagen II expression in Yougui Yin drug-containing serum + mechanical stimulation group. Conclusion: BMSCs can differentiate into chondrocytes on PLGA scaffold. Compared with blank serum group, BMSCs can be induced to differentiate into chondrocytes in all induction groups, among all induction groups, TGF-β1 + Yougui-yin drug-containing serum group and TGF-β1 + mechanical stimulation group were more significant, and the difference was statistically significant.展开更多
Background:Postmenopausal osteoporosis(PMO)is the most common primary osteoporosis in older women.This condition imposes a huge economic and medical burden on society.Aside from western medicine,traditional Chinese me...Background:Postmenopausal osteoporosis(PMO)is the most common primary osteoporosis in older women.This condition imposes a huge economic and medical burden on society.Aside from western medicine,traditional Chinese medicine is also widely used for the treatment of PMO,especially in Asian countries.Zuogui pill(ZGP)and Yougui pill(YGP)are classical formulas for the treatment of PMO with potent clinical effects.This study aimed to explore the potential active compounds,potential targets,and potential mechanisms of ZGP and YGP for the treatment of PMO.Methods:Compounds from ZGP and YGP were collected from three online databases,and these compounds were screened by oral bioavailability and drug-likeness.Their corresponding targets were determined from the Medicine Systems Pharmacology Database and Analysis platform.The corresponding targets for PMO were obtained from two disease databases.The target protein-protein interaction was identified through the STRING platforms and the core targets were ascertained by analyzing the protein-protein interaction network by using Cytoscape software.PMO-related genes were identified via Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses to identify specific biological processes,cellular components,molecular functions and key signalling pathways of ZGP and YGP for PMO treatment.Results:A total of 68 compounds were screened from ZGP with 168 putative potential target genes,whereas 99 compounds were screened from YGP with 214 potential target genes associated with PMO.Most of the core components included quercetin and kaempferol,and most of the core targets were TP53,AKT1,MAPK1,JUN,TNF,HSP90AA1,APP,IL6,VEGFA,RELA,MAPK8,NR3C1,EGFR,CXCL8,ESR1,FOS and MAPK14.Gene Ontology enrichment and Kyoto Encyclopedia of Genes and Genomes pathway analyses revealed that ZGP and YGP treat PMO primarily via regulation of bone formation and resorption,anti-inflammatory immunity and hormonal regulation.Conclusion:Our data provided insights into the mechanisms,by which ZGP and YGP treat PMO.This study points out a direction for further research into ZGP and YGP monomers and pathways and offers a new clinical treatment option for non-traditional Chinese medicine clinicians in the treatment of PMO.展开更多
Objective：To study the effects of Zuogui Pill（左归丸,ZGP）and Yougui Pill（右归丸,YGP）on the expressions of brain-derived neurotrophic factor（BDNF）and cyclic adenosine monophosphate（cAMP）/protein kinase A（PKA...Objective：To study the effects of Zuogui Pill（左归丸,ZGP）and Yougui Pill（右归丸,YGP）on the expressions of brain-derived neurotrophic factor（BDNF）and cyclic adenosine monophosphate（cAMP）/protein kinase A（PKA）signaling of axonal regeneration in the Lewis rats with experimental autoimmune encephalomyelitis（EAE）,in order to explore the possible mechanism of ZGP and YGP on promoting axonal regeneration.Methods：The rats were randomly divided into normal control（NC）,model（MO）,prednisone acetate（PA）,ZGP and YGP groups.The EAE model of rat was established by injecting antigen containing myelin basic protein（MBP）68-86.The brain and spinal cord were harvested on the 14th and 28th day postimmunization（PI）,the protein and mRNA expression of BDNF and PKA in the brain and spinal cord of rats were detected by Western blot analysis and real-time quantitative polymerase chain reaction（PCR）,and the cAMP levels were detected by using enzyme-immunoassay method.Results：（1）On the 28th day PI,the mRNA expression of BDNF in brain white matter and spinal cord of rats in ZGP and YGP groups were up-regulated,especially in YGP group（P〈0.05 or P〈0.01）.（2）On the 14th day PI,the cAMP levels in brain white matters significantly increased in PA and YGP groups compared with MO group（P〈0.05 or P〈0.01）,and the cAMP level in YGP group was higher than that in ZGP group（P〈0.05）.The cAMP level in spinal cord also significantly increased in YGP group compared with MO,PA and ZGP groups,respectively（P〈0.01）.（3）On the 14th day PI,the PKA expression in spinal cord of rats in ZGP group was significantly decreased compared with MO and YGP groups,respectively（P〈0.05）.（4）On the 28th day PI,there was a positive correlation between cAMP and PKA expression in the brain white matter of YGP rats.Conclusions：The results suggest that ZGP and YGP may promote axonal regeneration by modulating cAMP/PKA signal transduction pathway,but the targets of molecular mechanism of ZGP may be different from those of YGP.展开更多
Objective：To study the molecular mechanism of Zuogui Pill（左归丸,ZGP） and Yougui Pill（右归丸,YGP） on axonal regeneration in rats with experimental autoimmune encephalomyelitis（EAE）.Methods： EAE rat model was e...Objective：To study the molecular mechanism of Zuogui Pill（左归丸,ZGP） and Yougui Pill（右归丸,YGP） on axonal regeneration in rats with experimental autoimmune encephalomyelitis（EAE）.Methods： EAE rat model was established by bilateral rear pedes subcutaneous injection of antigen made by mixing myelin basic protein（MBP） and complete Freud＇s adjuvant（CFA） in the volume ratio of 1：1.The pathological changes of axonal injury and regeneration in the brain and the spinal cord were observed on the 14th（the acute stage） and the 28th day（the remission stage） after modeling,with hematoxylin-eosin（HE） staining,silver stain, and immunohistochemical staining.The rats treated with prednisone acetate were taken as controls.Results： Observation under the light microscope with HE staining showed a sleeve-like change in rats＇ cerebrospinal parenchyma with inflammatory cell infiltration around the small vessels and neuronic denaturation,while silver staining showed excessive tumefaction and abscission of axon,and immunohistochemical analysis showed decreasing of nerve growth factor（NGF） expression at the acute stage of EAE,which was even more remarkable at the remission stage,showing significant difference as compared with the normal control（P〈0.05）. And the expressions of Nogo A,an axon growth inhibitor,and its receptor（Nogo-66 receptor,Ng R） were significantly higher than those in the normal control at the acute stage（P〈0.01）.However,after the intervention of ZGP and YGP,the pathological changes and axon damage in rats＇ brain and spinal cord were much more alleviated,and the NGF expression was significantly higher than that in the model group at the acute stage （P〈0.05）.The expression of NGF was even stronger during the remission stage,and a better effect was shown by YGP.As for Nogo A and Ng R expressions,they were significantly lower than those in the model group at the acute stage（P〈0.05）,but a better effect was shown by ZGP.Conclusions：ZGP and YGP can prevent axonal injury and promote the axonal regeneration in rats of EAE,and the possible mechanism is to increase the expression of NGF and reduce the expression of Nogo A and its receptor.However,some differences are observed between the two Chinese preparations in their acting times and points,which provides a certain basis for revealing the modern connotation of the Chinese medicine theory on tonifying Shen（肾）-yin and Shen-yang.展开更多
基金Jiangxi University of Traditional Chinese Medicine Schoollevel Postgraduate Innovation Special Funds for Funding Projects(School Word[2023]No.33)Jiangxi Province Traditional Chinese Medicine Young and Middle-aged Backbone Talent Training Program(Third Batch)Project(Gan Traditional Chinese Medicine Science and Education[2021]No.4)。
文摘Objective This study aimed to explore the mechanism of obtaining yang from yin in Yougui pill against aging based on network pharmacology and molecular docking technology.Methods The active components and targets of Yougui Pill were obtained by searching the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)database and the Bioinformatics Analysis Tool for Molecular mechANism of Traditional Chinese Medicine(BATMEN-TCM)database,and kidney deficiency syndrome-related targets were obtained in the Symptom Mapping(SymMap)Database,a traditional Chinese medicine(TCM)syndrome correlation database.The protein–protein interaction(PPI)network was constructed by using the STRING11.5 database.Then,we used CytoScape3.9.0 software to construct the network of TCM–active components–potential targets,and the core TCM components and core targets of Yougui Pill for the treatment of kidney deficiency were obtained.The function analysis of Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathway enrichment analysis were performed in the Database for Annotation,Visualization,and Integrated Discovery(DAVID).Finally,preliminary verification was performed with the help of molecular docking technology.Results A total of 147 active components of 9 drugs of Yougui Pill(Fuzi[Aconiti Lateralis Radix Praeparata],Shudihaung[Rehmanniae Radix Praeparata],Gouqi[Lycii Fructus],Shanyao[Rhizoma Dioscoreae],Shanzhuyu[Corni Fructus],Tusizi[Cuscutae Semen],Danggui[Angelicae Sinensis Radix],Duzhong[Eucommiae Cortex],Lujiaojiao[Cervi Cornus Colla])were obtained,corresponding to 233 targets.A total of 2,235targets related to kidney deficiency syndrome and 43 potential therapeutic targets were obtained after the intersection.The core TCM components mainly included quercetin,kaempferol,diosgenin,β-carotene,etc.The core targets involved Trp53(Tp53),Akt1,Pparg,Nr3c1,App,Casp8,Mapk1,Cav1,and Ctnnb1.A total of 27biological processes,10 cellular components,and 11 molecular functions were obtained by gene function enrichment analysis,mainly related to the regulation of gene expression,cell apoptosis and proliferation,and the response to estrogen.A total of 51 KEGG signaling pathways,mainly involving a variety of cancer pathways,apoptosis pathways,longevity regulation pathways,etc.Conclusion Yougui Pill can play a role in preventing and treating kidney deficiency syndrome through multiple targets and pathways.
基金Supported by the National Natural Science Foundation of China Project:Study on the Biological Basis of the Classic Prescription Erzhi pills for Tonifying the Liver and Kidney(No.81630106)。
文摘OBJECTIVE:To examine,dissect,and understand the molecular mechanisms and combinatorial effects of Zuogui(左归丸,ZGP)and Yougui pills(右归丸,YGP)in 4-vinyl cyclohexene diepoxide(4-VCD)-induced Perimenopausal syndrome(PMS).METHODS:Using the 4-VCD-induced PMS mouse model,uterine and ovary index were measured,and serum sex steroidal hormone levels were evaluated after treatment with ZGP,YGP,ZGP+YGP,estradiol valerate(EV),and Gengnian An(更年安,GNA).Histopathological examinations,ingredient-target network predictions,western blotting,and real-time quantitative polymerase chain reaction(RT-q PCR)analyses were performed to ascertain the possible pharmacological effects and molecular mechanisms of ZYP and YGP.RESULTS:Treatment with ZGP and YGP remarkably improves estrous cyclicity and prevents pathological damage to the uterus.Also,altered sex hormones,including AMH,E2,FSH,LH,P,and T,were restored to normal levels after ZGP and YGP administration.Ingredient-target network analysis showed that the 5 ingredients common to the ZGP and YGP formula modulate 53 targets shared with PMS.Pathwayenrichment analysis further predicted that ZGY and YGP likely regulate of apoptosis and other essential pathways during PMS.In-vivo studies showed that ZGP and YGP suppress PMS via modulating apoptosis through decreasing Caspase-3 and Bcl-2-associated X(Bax)levels and increasing B-cell lymphoma-2(Bcl-2)/Bax and Bcl-2 levels.Importantly,ZGP+YGP treatment modulation effects were somewhat or significantly better compared to ZGP or YGP alone treatment.CONCLUSION:ZGP and YGP represent novel anti-PMS agents whose effects involve restoring altered hormonal levels,protecting the uterus,and regulating apoptosis.
基金Supported by the National Natural Science Foundation of China,No.81973754Shaanxi University of Traditional Chinese Medicine Innovation Team Project,No.2019-QN06.
文摘BACKGROUND Yougui pills have long been used to treat hypothyroidism,usually in combination with levothyroxine sodium in clinical treatment,while their clinical efficacy and safety are still controversial when compared to levothyroxine treatment alone.AIM To explore the clinical efficacy and safety of Yougui pills combined with levothyroxine sodium in the treatment of hypothyroidism.METHODS This meta-analysis was performed in accordance with the PRISMA guidelines.Randomized controlled trials on Yougui pills in the treatment of hypothyroidism published from 2008 to May 2021 were searched in a total of 8 databases(4 databases in Chinese and 4 databases in English).The quality of the included studies was evaluated according to the Cochrane risk assessment tool.Weighted mean difference(WMD)was used for continuous variables,and relative risk(RR)was used for binary variables.Data were extracted,and the meta-analysis was conducted with the statistical software of Stata15.0 and RevMan5.0.RESULTS A total of 140 articles were retrieved,and 9 of them were finally included,with a total sample size of 936 cases.The main meta-analysis results are as follows:(1)The group of Yougui pills combined with levothyroxine sodium had a significantly higher overall response rate than the group of levothyroxine sodium(RR=1.20,95%CI 1.12,1.28,P<0.00001);(2)Yougui pills combined with levothyroxine sodium achieved significantly better efficacy than levothyroxine sodium alone in alleviating adverse symptoms[standard mean difference(SMD)=-1.10,95%CI:-1.37,-0.84,P<0.00001];(3)The level of thyrotropin stimulating hormone in the group of Yougui pills combined with levothyroxine sodium was significantly lower than in the control group of levothyroxine sodium(WMD=-1.38,95%CI:-2.10,-0.67,P=0.00001);(4)The level of free triiodothyronine in the group of Yougui pills combined with levothyroxine sodium was higher than that in the control group of levothyroxine sodium(WMD=0.41,95%CI:0.03,0.79,P=0.03);(5)The level of free thyroxine in the group of Yougui pills combined with levothyroxine sodium was significantly higher than that in the control group of levothyroxine sodium(SMD=0.83,95%CI:0.44,1.22,P≤0.0001);and(6)The adverse reactions in the group of Yougui pills combined with levothyroxine sodium were significantly less than those in the control group of levothyroxine sodium(RR=0.33,95%CI:0.20,-0.53,P<0.00001).CONCLUSION In the treatment of hypothyroidism,the combination of Yougui pills with levothyroxine sodium may be better than levothyroxine sodium treatment alone.
基金supported by the National Natural Science Foundation of China,No.30471576,and 30872277a grant from the Innovative Research Program in Universities of the Ministry of Education of China,No.IRT0760
文摘Long-term glucocorticoid use may result in sustained suppression of one or more secreted components from the hypothalamo-pituitary-adrenal axis, and often results in apoptosis. Yougui Pill（YGP）, a 10-component traditional Chinese herbal medicine, has been shown to be clinically effective for glucocorticoid-induced suppression of the hypothalamo-pituitary-adrenal axis. However, the pharmacological and molecular mechanisms remain unclear. We hypothesized that YGP would exert an anti-apoptosis effect on dexamethasone-treated anterior pituitary cells. In vivo experiments showed that YGP significantly reduced the number of apoptotic cells, down-regulated m RNA expression of cytochrome c, caspase-3, and caspase-9, and up-regulated m RNA expression of Bcl-2. These findings suggest that YGP reduced glucocorticoid-induced apoptosis in rat anterior pituitary cells by regulating the mitochondria-mediated apoptosis pathway.
文摘Objective: Bone marrow mesenchymal stem cells (BMSCs) have the potential to differentiate into chondrocytes, it is the ideal seed cells for treating cartilage injury. The purpose of this study was to investigate the inducible factors that stimulate the differentiation of bone marrow mesenchymal stem cells into chondrocytes. Methods: Rat BMSCs were isolated and subcultured, and then inoculated on PLGA scaffolds after successfully identificating of BMSCs. They were divided into 5 groups. Blank serum group, TGF-β1 + Yougui Yin drug-containing serum induction group, TGF-β1 + mechanical stimulation induction group, Yougui Yin drug-containing serum + mechanical stimulation induction group, TGF-β1 + Yougui Yin drug-containing serum + mechanical stimulation induction group, 4 weeks later, the adhesion status of cells and scaffolds was observed by Scanning Electron Microscopy, and then chondrocyte induction was performed on different treatment groups. After induction, the expression of Aggrecan and Collagen II was detected by immunofluorescence, and the mRNA expression of Aggrecan and Collagen II was detected by qPCR. Results: BMSCs were identified as rat bone marrow mesenchymal stem cells by immunofluorescence CD34 and CD44 cells. Scanning Electron Microscopy observed that BMSCs grew well on PLGA. Compared with PLGA group, the scaffold space of BMSCS-PLGA composite was significantly reduced, and the growth of cell adhesion scaffold was significantly increased. Compared with the blank serum group, the expression of Aggrecan in TGF-β1 + Yougui Yin drug-containing serum group and Yougui Yin drug-containing serum + mechanical stimulation group was significantly increased. The expression of Collagen II in TGF-β1 + Yougui Yin medicated serum group, Yougui Yin medicated serum + mechanical stimulation group, TGF-β1 + mechanical stimulation group and TGF-β1 + Yougui Yin medicated serum + mechanical stimulation group were significantly increased;PCR results showed that compared with blank serum group, the expression of Aggrecan in other groups was significantly increased, except for Collagen II expression in Yougui Yin drug-containing serum + mechanical stimulation group. Conclusion: BMSCs can differentiate into chondrocytes on PLGA scaffold. Compared with blank serum group, BMSCs can be induced to differentiate into chondrocytes in all induction groups, among all induction groups, TGF-β1 + Yougui-yin drug-containing serum group and TGF-β1 + mechanical stimulation group were more significant, and the difference was statistically significant.
基金supported by Science and Technology Commission of Shanghai Municipality,China(20Z21900400).
文摘Background:Postmenopausal osteoporosis(PMO)is the most common primary osteoporosis in older women.This condition imposes a huge economic and medical burden on society.Aside from western medicine,traditional Chinese medicine is also widely used for the treatment of PMO,especially in Asian countries.Zuogui pill(ZGP)and Yougui pill(YGP)are classical formulas for the treatment of PMO with potent clinical effects.This study aimed to explore the potential active compounds,potential targets,and potential mechanisms of ZGP and YGP for the treatment of PMO.Methods:Compounds from ZGP and YGP were collected from three online databases,and these compounds were screened by oral bioavailability and drug-likeness.Their corresponding targets were determined from the Medicine Systems Pharmacology Database and Analysis platform.The corresponding targets for PMO were obtained from two disease databases.The target protein-protein interaction was identified through the STRING platforms and the core targets were ascertained by analyzing the protein-protein interaction network by using Cytoscape software.PMO-related genes were identified via Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses to identify specific biological processes,cellular components,molecular functions and key signalling pathways of ZGP and YGP for PMO treatment.Results:A total of 68 compounds were screened from ZGP with 168 putative potential target genes,whereas 99 compounds were screened from YGP with 214 potential target genes associated with PMO.Most of the core components included quercetin and kaempferol,and most of the core targets were TP53,AKT1,MAPK1,JUN,TNF,HSP90AA1,APP,IL6,VEGFA,RELA,MAPK8,NR3C1,EGFR,CXCL8,ESR1,FOS and MAPK14.Gene Ontology enrichment and Kyoto Encyclopedia of Genes and Genomes pathway analyses revealed that ZGP and YGP treat PMO primarily via regulation of bone formation and resorption,anti-inflammatory immunity and hormonal regulation.Conclusion:Our data provided insights into the mechanisms,by which ZGP and YGP treat PMO.This study points out a direction for further research into ZGP and YGP monomers and pathways and offers a new clinical treatment option for non-traditional Chinese medicine clinicians in the treatment of PMO.
基金Supported by the National Natural Science Foundation of China(No.30873230)Beijing Natural Science Foundation(No.7092014)+1 种基金Scientific Research Common Program of Beijing Municipal Education Commission(No.KM2007100025015)Funding Project for Academic Human Resources Development in Institutions of Higher Learning Under the Jurisdiction of Beijing Municipality(No.PHR201008401)
文摘Objective：To study the effects of Zuogui Pill（左归丸,ZGP）and Yougui Pill（右归丸,YGP）on the expressions of brain-derived neurotrophic factor（BDNF）and cyclic adenosine monophosphate（cAMP）/protein kinase A（PKA）signaling of axonal regeneration in the Lewis rats with experimental autoimmune encephalomyelitis（EAE）,in order to explore the possible mechanism of ZGP and YGP on promoting axonal regeneration.Methods：The rats were randomly divided into normal control（NC）,model（MO）,prednisone acetate（PA）,ZGP and YGP groups.The EAE model of rat was established by injecting antigen containing myelin basic protein（MBP）68-86.The brain and spinal cord were harvested on the 14th and 28th day postimmunization（PI）,the protein and mRNA expression of BDNF and PKA in the brain and spinal cord of rats were detected by Western blot analysis and real-time quantitative polymerase chain reaction（PCR）,and the cAMP levels were detected by using enzyme-immunoassay method.Results：（1）On the 28th day PI,the mRNA expression of BDNF in brain white matter and spinal cord of rats in ZGP and YGP groups were up-regulated,especially in YGP group（P〈0.05 or P〈0.01）.（2）On the 14th day PI,the cAMP levels in brain white matters significantly increased in PA and YGP groups compared with MO group（P〈0.05 or P〈0.01）,and the cAMP level in YGP group was higher than that in ZGP group（P〈0.05）.The cAMP level in spinal cord also significantly increased in YGP group compared with MO,PA and ZGP groups,respectively（P〈0.01）.（3）On the 14th day PI,the PKA expression in spinal cord of rats in ZGP group was significantly decreased compared with MO and YGP groups,respectively（P〈0.05）.（4）On the 28th day PI,there was a positive correlation between cAMP and PKA expression in the brain white matter of YGP rats.Conclusions：The results suggest that ZGP and YGP may promote axonal regeneration by modulating cAMP/PKA signal transduction pathway,but the targets of molecular mechanism of ZGP may be different from those of YGP.
基金Supported by the National Natural Science Foundation of China (No.30873230)Beijing Natural Science Foundation(No. 7062022,No.7092014)
文摘Objective：To study the molecular mechanism of Zuogui Pill（左归丸,ZGP） and Yougui Pill（右归丸,YGP） on axonal regeneration in rats with experimental autoimmune encephalomyelitis（EAE）.Methods： EAE rat model was established by bilateral rear pedes subcutaneous injection of antigen made by mixing myelin basic protein（MBP） and complete Freud＇s adjuvant（CFA） in the volume ratio of 1：1.The pathological changes of axonal injury and regeneration in the brain and the spinal cord were observed on the 14th（the acute stage） and the 28th day（the remission stage） after modeling,with hematoxylin-eosin（HE） staining,silver stain, and immunohistochemical staining.The rats treated with prednisone acetate were taken as controls.Results： Observation under the light microscope with HE staining showed a sleeve-like change in rats＇ cerebrospinal parenchyma with inflammatory cell infiltration around the small vessels and neuronic denaturation,while silver staining showed excessive tumefaction and abscission of axon,and immunohistochemical analysis showed decreasing of nerve growth factor（NGF） expression at the acute stage of EAE,which was even more remarkable at the remission stage,showing significant difference as compared with the normal control（P〈0.05）. And the expressions of Nogo A,an axon growth inhibitor,and its receptor（Nogo-66 receptor,Ng R） were significantly higher than those in the normal control at the acute stage（P〈0.01）.However,after the intervention of ZGP and YGP,the pathological changes and axon damage in rats＇ brain and spinal cord were much more alleviated,and the NGF expression was significantly higher than that in the model group at the acute stage （P〈0.05）.The expression of NGF was even stronger during the remission stage,and a better effect was shown by YGP.As for Nogo A and Ng R expressions,they were significantly lower than those in the model group at the acute stage（P〈0.05）,but a better effect was shown by ZGP.Conclusions：ZGP and YGP can prevent axonal injury and promote the axonal regeneration in rats of EAE,and the possible mechanism is to increase the expression of NGF and reduce the expression of Nogo A and its receptor.However,some differences are observed between the two Chinese preparations in their acting times and points,which provides a certain basis for revealing the modern connotation of the Chinese medicine theory on tonifying Shen（肾）-yin and Shen-yang.
