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。
