AIM To explore the protective effects and underlying mechanisms of total polysaccharides of the Sijunzi decoction(TPSJ) on the epithelial barriers in vitro. METHODS Caco-2 cell monolayers were treated with or without ...AIM To explore the protective effects and underlying mechanisms of total polysaccharides of the Sijunzi decoction(TPSJ) on the epithelial barriers in vitro. METHODS Caco-2 cell monolayers were treated with or without TPSJ in the presence or absence of TNF-α, and paracellular permeability and transepithelial electrical resistance(TEER) were measured to evaluate the epithelial barrier function. Immunofluorescence and western blotting were respecti-vely used to evaluate the distribution and expression of the tight junction proteins claudin 1, claudin 2, zo3, and occludin in Caco-2 cells. western blotting was also used to evaluate the cellular expression of myosin light chain(MLC), phosphorylated MLC(pM LC), MLC kinase(MLCK), and nuclear factor(NF)-κB p65. RESULTS TPSJ promoted the proliferation of Caco-2 cells and inhibited TNF-α-induced secretion of pro-inflammatory cyto-kines. Furthermore, TPSJ significantly ameliorated both the reduction of TEER and the increased paracellular permeability observed in tumor necrosis factor(TNF)-α-damaged Caco-2 monolayers. Furthermore, TPSJ remarkably attenuated TNF-α-induced morphological changes, downregulated the expression of claudin 1, claudin 2, zo3, and occludin, and markedly suppressed TNF-α-mediated upregulation of p-MLC and MLCK expression. Finally, TPSJ inhibited the activation and expression of NF-κB p65. CONCLUSION Our results demonstrate that TPSJ alleviates the TNF-α-induced impairment of the intestinal epithelial cell barrier function by suppressing NF-κB p65-mediated phosphorylation of MLCK and MLC.展开更多
[目的]运用网络药理学联合生物信息学方法,探究黄芪四君子汤治疗肝细胞癌(hepatocelluar carcinoma,HCC)的潜在作用机制。[方法]利用中药系统药理学数据库和分析平台(Traditional Chinese Medicine Systems Pharmacology Database and A...[目的]运用网络药理学联合生物信息学方法,探究黄芪四君子汤治疗肝细胞癌(hepatocelluar carcinoma,HCC)的潜在作用机制。[方法]利用中药系统药理学数据库和分析平台(Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,TCMSP)数据库对中药的生物活性成分及其靶点进行鉴定。采用差异分析和加权基因共表达网络分析(weighted gene coexpression net work analysis,WGCNA)鉴定HCC的差异基因和模块基因,在此基础上构建中药成分-靶点网络。随后研究靶点的生物学功能,并构建蛋白互作(protein-protein interaction,PPI)网络,识别药物治疗HCC的关键靶点。最后通过分子对接探索化合物与靶点之间的相互作用。[结果]共鉴定出5种草药的156种化合物和227个靶点,癌症基因组图谱(The Cancer Genome Atlas,TCGA)和基因表达综合(Gene Expression Omnibus,GEO)数据库数据集中差异基因2477和685个,WGCNA关联模块基因2104和2165个。构建了包含4种草药、85种化合物和9个靶点的中药成分-靶点网络。生物功能分析表明,9个靶点主要与细胞周期、p53信号通路、细胞衰老相关,筛选出山柰酚、槲皮素2个药物主要成分和细胞周期蛋白A2(cyclin A2,CCNA2)、雌激素受体1(estrogen receptor alpha,ESR1)、细胞周期蛋白B1(cyclin B1,CCNB1)、细胞周期蛋白依赖性激酶1(cyclin-dependent kinase 1,CDK1)、拓扑异构酶Ⅱα(topoisomeraseⅡalpha,TOP2A)5个核心靶点。分子对接结果显示,山柰酚、槲皮素与对应的核心靶点具有稳定的结合能力。[结论]黄芪四君子汤可能通过调节细胞周期蛋白以及相关通路发挥对HCC的治疗作用,本研究为黄芪四君子汤治疗HCC提供了研究思路和理论支撑。展开更多
目的初步探讨四君子汤复方总多糖对小鼠肠上皮间淋巴细胞(intestinal intraepithelial lymphocyte,iIEL)免疫功能的影响。方法取NIH小鼠,按体重随机分为正常组、模型组、四君子汤复方总多糖(Polysaccharides of Si-Jun-Zi Decoction,SJZ...目的初步探讨四君子汤复方总多糖对小鼠肠上皮间淋巴细胞(intestinal intraepithelial lymphocyte,iIEL)免疫功能的影响。方法取NIH小鼠,按体重随机分为正常组、模型组、四君子汤复方总多糖(Polysaccharides of Si-Jun-Zi Decoction,SJZPS)组和四君子汤去蛋白多糖(Free protein of SJZPS,SJZFP)组,非连续密度梯度离心法提取iIEL,流式细胞(Flow Cytometry,FCM)技术检测iIEL中CD3+和CD69+细胞比例,ELISA法检测iIEL上清液的IFN-γ和IL-2浓度。结果iIEL中CD3+细胞比例为88.46%;环磷酰胺可使iIEL中CD69+细胞比例下降,iIEL上清液中IFN-γ和IL-2浓度降低,与正常组比较,具有非常显著性差异(P<0.01);SJZPS和SJZFP组中iIEL的CD69+细胞比例以及IFN-γ和IL-2分泌水平明显升高,与模型组比较,具有显著性差异(P<0.05或P<0.01)。结论SJZPS和SJZFP均可提高环磷酰胺抑制的小鼠肠上皮间淋巴细胞的免疫功能,具有肠道黏膜免疫调节作用。展开更多
基于网络药理学探讨四君子汤治疗阿尔茨海默病(AD)的作用机制。借助TCMSP数据库及Uniprot数据库筛选出四君子汤有效成分及靶点基因。通过Drugbank、Dis Ge NET和TTD数据库筛选出阿尔茨海默病的靶点基因;成分靶点与疾病靶点映射后使用Cyt...基于网络药理学探讨四君子汤治疗阿尔茨海默病(AD)的作用机制。借助TCMSP数据库及Uniprot数据库筛选出四君子汤有效成分及靶点基因。通过Drugbank、Dis Ge NET和TTD数据库筛选出阿尔茨海默病的靶点基因;成分靶点与疾病靶点映射后使用Cytoscape 3.7.1软件构建药物有效成分-靶点蛋白相互作用网络,使用String数据库绘制靶点蛋白-靶点蛋白相互作用网络;对靶点蛋白利用Metascape数据库进行GO分析和KEGG分析,最后采用荧光实时定量PCR对网络药理学主要分析结果进行验证。分析结果表明,四君子汤主要关联AD的β淀粉样蛋白聚集、细胞凋亡、炎症反应、氧化应激反应、自噬、胰岛素代谢等;体外实验提示,四君子汤参与调控APP介导的β淀粉样蛋白聚集,Caspase-3、Bcl-2、Bax介导的细胞凋亡,MAOB介导的氧化应激反应,mTOR介导的自噬,以及INSR介导的胰岛素代谢等通路。综上四君子汤治疗AD具有多成分、多靶点的特点,可为进一步研究其作用机制提供依据。展开更多
基金Supported by the National Natural Science Foundation of China,No.81202635the Guangdong Provincial Bureau of Chinese Medicine,No.20151244
文摘AIM To explore the protective effects and underlying mechanisms of total polysaccharides of the Sijunzi decoction(TPSJ) on the epithelial barriers in vitro. METHODS Caco-2 cell monolayers were treated with or without TPSJ in the presence or absence of TNF-α, and paracellular permeability and transepithelial electrical resistance(TEER) were measured to evaluate the epithelial barrier function. Immunofluorescence and western blotting were respecti-vely used to evaluate the distribution and expression of the tight junction proteins claudin 1, claudin 2, zo3, and occludin in Caco-2 cells. western blotting was also used to evaluate the cellular expression of myosin light chain(MLC), phosphorylated MLC(pM LC), MLC kinase(MLCK), and nuclear factor(NF)-κB p65. RESULTS TPSJ promoted the proliferation of Caco-2 cells and inhibited TNF-α-induced secretion of pro-inflammatory cyto-kines. Furthermore, TPSJ significantly ameliorated both the reduction of TEER and the increased paracellular permeability observed in tumor necrosis factor(TNF)-α-damaged Caco-2 monolayers. Furthermore, TPSJ remarkably attenuated TNF-α-induced morphological changes, downregulated the expression of claudin 1, claudin 2, zo3, and occludin, and markedly suppressed TNF-α-mediated upregulation of p-MLC and MLCK expression. Finally, TPSJ inhibited the activation and expression of NF-κB p65. CONCLUSION Our results demonstrate that TPSJ alleviates the TNF-α-induced impairment of the intestinal epithelial cell barrier function by suppressing NF-κB p65-mediated phosphorylation of MLCK and MLC.
文摘[目的]运用网络药理学联合生物信息学方法,探究黄芪四君子汤治疗肝细胞癌(hepatocelluar carcinoma,HCC)的潜在作用机制。[方法]利用中药系统药理学数据库和分析平台(Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,TCMSP)数据库对中药的生物活性成分及其靶点进行鉴定。采用差异分析和加权基因共表达网络分析(weighted gene coexpression net work analysis,WGCNA)鉴定HCC的差异基因和模块基因,在此基础上构建中药成分-靶点网络。随后研究靶点的生物学功能,并构建蛋白互作(protein-protein interaction,PPI)网络,识别药物治疗HCC的关键靶点。最后通过分子对接探索化合物与靶点之间的相互作用。[结果]共鉴定出5种草药的156种化合物和227个靶点,癌症基因组图谱(The Cancer Genome Atlas,TCGA)和基因表达综合(Gene Expression Omnibus,GEO)数据库数据集中差异基因2477和685个,WGCNA关联模块基因2104和2165个。构建了包含4种草药、85种化合物和9个靶点的中药成分-靶点网络。生物功能分析表明,9个靶点主要与细胞周期、p53信号通路、细胞衰老相关,筛选出山柰酚、槲皮素2个药物主要成分和细胞周期蛋白A2(cyclin A2,CCNA2)、雌激素受体1(estrogen receptor alpha,ESR1)、细胞周期蛋白B1(cyclin B1,CCNB1)、细胞周期蛋白依赖性激酶1(cyclin-dependent kinase 1,CDK1)、拓扑异构酶Ⅱα(topoisomeraseⅡalpha,TOP2A)5个核心靶点。分子对接结果显示,山柰酚、槲皮素与对应的核心靶点具有稳定的结合能力。[结论]黄芪四君子汤可能通过调节细胞周期蛋白以及相关通路发挥对HCC的治疗作用,本研究为黄芪四君子汤治疗HCC提供了研究思路和理论支撑。
文摘目的初步探讨四君子汤复方总多糖对小鼠肠上皮间淋巴细胞(intestinal intraepithelial lymphocyte,iIEL)免疫功能的影响。方法取NIH小鼠,按体重随机分为正常组、模型组、四君子汤复方总多糖(Polysaccharides of Si-Jun-Zi Decoction,SJZPS)组和四君子汤去蛋白多糖(Free protein of SJZPS,SJZFP)组,非连续密度梯度离心法提取iIEL,流式细胞(Flow Cytometry,FCM)技术检测iIEL中CD3+和CD69+细胞比例,ELISA法检测iIEL上清液的IFN-γ和IL-2浓度。结果iIEL中CD3+细胞比例为88.46%;环磷酰胺可使iIEL中CD69+细胞比例下降,iIEL上清液中IFN-γ和IL-2浓度降低,与正常组比较,具有非常显著性差异(P<0.01);SJZPS和SJZFP组中iIEL的CD69+细胞比例以及IFN-γ和IL-2分泌水平明显升高,与模型组比较,具有显著性差异(P<0.05或P<0.01)。结论SJZPS和SJZFP均可提高环磷酰胺抑制的小鼠肠上皮间淋巴细胞的免疫功能,具有肠道黏膜免疫调节作用。
文摘基于网络药理学探讨四君子汤治疗阿尔茨海默病(AD)的作用机制。借助TCMSP数据库及Uniprot数据库筛选出四君子汤有效成分及靶点基因。通过Drugbank、Dis Ge NET和TTD数据库筛选出阿尔茨海默病的靶点基因;成分靶点与疾病靶点映射后使用Cytoscape 3.7.1软件构建药物有效成分-靶点蛋白相互作用网络,使用String数据库绘制靶点蛋白-靶点蛋白相互作用网络;对靶点蛋白利用Metascape数据库进行GO分析和KEGG分析,最后采用荧光实时定量PCR对网络药理学主要分析结果进行验证。分析结果表明,四君子汤主要关联AD的β淀粉样蛋白聚集、细胞凋亡、炎症反应、氧化应激反应、自噬、胰岛素代谢等;体外实验提示,四君子汤参与调控APP介导的β淀粉样蛋白聚集,Caspase-3、Bcl-2、Bax介导的细胞凋亡,MAOB介导的氧化应激反应,mTOR介导的自噬,以及INSR介导的胰岛素代谢等通路。综上四君子汤治疗AD具有多成分、多靶点的特点,可为进一步研究其作用机制提供依据。
