目的:探讨脂多糖(lipopolysacoharides,LPS)对胆管癌细胞上皮间质转化(epithelial-mesenchymaltransition,EMT)的影响和可能机制.方法:将胆管癌ICBD细胞分为4组:正常对照组、LPS诱导实验组(终浓度10g/mL)、LPS+siRNA转染组和LPS+SB-203...目的:探讨脂多糖(lipopolysacoharides,LPS)对胆管癌细胞上皮间质转化(epithelial-mesenchymaltransition,EMT)的影响和可能机制.方法:将胆管癌ICBD细胞分为4组:正常对照组、LPS诱导实验组(终浓度10g/mL)、LPS+siRNA转染组和LPS+SB-203580诱导实验组.应用Real-time RT-PCR与Westernb l o t法检测上皮细胞表面标志E-钙黏蛋白(E-cadherin)和间质细胞表面标志波形蛋白(Vimentin)的表达变化以及Toll样受体4(Toll-like receptors4,TLR4)和p38的表达变化.结果:LPS促进胆管癌细胞系ICBD的EMT发生;ICBD细胞的EMT过程伴随TLR4、p38表达增加;应用siRNA阻断TLR4后,ICBD细胞的EMT消失,LPS导致p38的上调表达作用也消失;应用SB-203580阻断p38后,与正常对照组相比,TLR4的表达增加,与LPS诱导实验组相比无明显变化,但ICBD细胞的EMT消失.结论:LPS可以激活TLR4,并通过p38/MAPK促进胆管癌细胞ICBD的上皮间质转化.展开更多
AIM To investigate the influence of high salt on dextran sulfate sodium(DSS)-induced colitis in mice and explore the underlying mechanisms of this effect.METHODS DSS and NaC l were used to establish the proinflammator...AIM To investigate the influence of high salt on dextran sulfate sodium(DSS)-induced colitis in mice and explore the underlying mechanisms of this effect.METHODS DSS and NaC l were used to establish the proinflammatory animal model. We evaluated the colitis severity. Flow cytometry was employed for detecting the frequencies of Th1, macrophages and Tregs in spleen, mesenteric lymph node and lamina propria. The important role of macrophages in the promotion of DSS-induced colitis by NaCl was evaluated by depleting macrophages with clodronate liposomes. Activated peritoneal macrophages and lamina propria mononuclear cells(LPMCs) were stimulated with NaCl, and proteins were detected by western blotting. Cytokines and inflammation genes were analyzed by enzyme-linked immunosorbent assay and RT-PCR, respectively.RESULTS The study findings indicate that NaC l up-regulates the frequencies of CD11b^+ macrophages and CD4^+IFN-γ^+IL-17^+ T cells in lamina propria in DSS-treated mice. CD3^+CD4^+CD25^+Foxp^3+ T cells, which can secrete high levels of IL-10 and TGF-β, increase through feedback in NaCl-and DSS-treated mice. Furthermore, clodronate liposomes pretreatment significantly alleviated DSSinduced colitis, indicating that macrophages play a vital role in NaCl proinflammatory activity. NaCl aggravates peritoneal macrophage inflammation by promoting the expressions of interleukin(IL)-1, IL-6 and mouse inducible nitric oxide synthase. Specifically, high NaCl concentrations promote p38 phosphorylation in lipopolysaccharide-and IFN-γ-activated LPMCs mediated by SGK1. CONCLUSION Proinflammatory macrophages may play an essential role in the onset and development of NaCl-promoted inflammation in DSS-induced colitis. The underlining mechanism involves up-regulation of the p38/MAPK axis.展开更多
文摘目的:探讨脂多糖(lipopolysacoharides,LPS)对胆管癌细胞上皮间质转化(epithelial-mesenchymaltransition,EMT)的影响和可能机制.方法:将胆管癌ICBD细胞分为4组:正常对照组、LPS诱导实验组(终浓度10g/mL)、LPS+siRNA转染组和LPS+SB-203580诱导实验组.应用Real-time RT-PCR与Westernb l o t法检测上皮细胞表面标志E-钙黏蛋白(E-cadherin)和间质细胞表面标志波形蛋白(Vimentin)的表达变化以及Toll样受体4(Toll-like receptors4,TLR4)和p38的表达变化.结果:LPS促进胆管癌细胞系ICBD的EMT发生;ICBD细胞的EMT过程伴随TLR4、p38表达增加;应用siRNA阻断TLR4后,ICBD细胞的EMT消失,LPS导致p38的上调表达作用也消失;应用SB-203580阻断p38后,与正常对照组相比,TLR4的表达增加,与LPS诱导实验组相比无明显变化,但ICBD细胞的EMT消失.结论:LPS可以激活TLR4,并通过p38/MAPK促进胆管癌细胞ICBD的上皮间质转化.
基金Supported by National Natural Science Foundation of China,No.81271813 and No.81570497
文摘AIM To investigate the influence of high salt on dextran sulfate sodium(DSS)-induced colitis in mice and explore the underlying mechanisms of this effect.METHODS DSS and NaC l were used to establish the proinflammatory animal model. We evaluated the colitis severity. Flow cytometry was employed for detecting the frequencies of Th1, macrophages and Tregs in spleen, mesenteric lymph node and lamina propria. The important role of macrophages in the promotion of DSS-induced colitis by NaCl was evaluated by depleting macrophages with clodronate liposomes. Activated peritoneal macrophages and lamina propria mononuclear cells(LPMCs) were stimulated with NaCl, and proteins were detected by western blotting. Cytokines and inflammation genes were analyzed by enzyme-linked immunosorbent assay and RT-PCR, respectively.RESULTS The study findings indicate that NaC l up-regulates the frequencies of CD11b^+ macrophages and CD4^+IFN-γ^+IL-17^+ T cells in lamina propria in DSS-treated mice. CD3^+CD4^+CD25^+Foxp^3+ T cells, which can secrete high levels of IL-10 and TGF-β, increase through feedback in NaCl-and DSS-treated mice. Furthermore, clodronate liposomes pretreatment significantly alleviated DSSinduced colitis, indicating that macrophages play a vital role in NaCl proinflammatory activity. NaCl aggravates peritoneal macrophage inflammation by promoting the expressions of interleukin(IL)-1, IL-6 and mouse inducible nitric oxide synthase. Specifically, high NaCl concentrations promote p38 phosphorylation in lipopolysaccharide-and IFN-γ-activated LPMCs mediated by SGK1. CONCLUSION Proinflammatory macrophages may play an essential role in the onset and development of NaCl-promoted inflammation in DSS-induced colitis. The underlining mechanism involves up-regulation of the p38/MAPK axis.
