Shi Haitao, Dong Lei, Liu Yaping, Jia Miao, Zhao Gang, Zhao Juhui, Lu Xiaolan
Department of Gastroenterology, Second Affiliated Hospital of Xian Jiaotong University College of Medicine, Xi'an 710004, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2013 Mar;33(3):386-90.
To investigate the effect of high glucose on the expressions of toll-like receptor 4 (TLR4) and proinflammatory cytokine production induced by lipopolysaccharide (LPS) in hepatic stellate cells in vitro.
Hepatic stellate cell line T6 was cultured in vitro and stimulated by high glucose. The mRNA and protein expression of TLR4 were detected by RT-PCR and Western blotting, respectively. After a 24-h pretreatment with high or low glucose, the cells were stimulated with LPS for 2 h, and Western blotting was used to detect the nuclear translocation of nuclear factor-κB (NF-κB); at 24 h of LPS exposure, the cells were examined for MCP-1 and IL-6 mRNA and protein expression levels with RT-PCR and ELISA, respectively.
High glucose significantly increased the mRNA and protein expressions of TLR4 (P<0.01) in a time- and dose-dependent manner. High glucose promoted NF-κB nuclear translocation and significantly enhanced the expression and secretion of both MCP-1 and IL-6 (P<0.01). Pretreatment with high glucose significantly promoted LPS-induced NF-κB nuclear translocation (P<0.01) and the mRNA expression and secretion of MCP-1 and IL-6.
High glucose can increase TLR4 mRNA and protein expressions in hepatic stellate cells and promote LPS-induced NF-κB activation and production of proinflammatory cytokines.
探讨高糖对体外培养的肝星状细胞中Toll样受体4（TLR4）表达及脂多糖（LPS）诱导的促炎细胞因子产生的影响。
体外培养肝星状细胞系T6并用高糖刺激。分别采用逆转录-聚合酶链反应（RT-PCR）和蛋白质印迹法检测TLR4的mRNA和蛋白表达。用高糖或低糖预处理细胞24小时后，用LPS刺激细胞2小时，采用蛋白质印迹法检测核因子-κB（NF-κB）的核转位；在LPS作用24小时时，分别用RT-PCR和酶联免疫吸附测定（ELISA）检测细胞中单核细胞趋化蛋白-1（MCP-1）和白细胞介素-6（IL-6）的mRNA和蛋白表达水平。
高糖以时间和剂量依赖的方式显著增加TLR4的mRNA和蛋白表达（P<0.01）。高糖促进NF-κB核转位，并显著增强MCP-1和IL-6的表达及分泌（P<0.01）。高糖预处理显著促进LPS诱导的NF-κB核转位（P<0.01）以及MCP-1和IL-6的mRNA表达及分泌。
高糖可增加肝星状细胞中TLR4的mRNA和蛋白表达，并促进LPS诱导的NF-κB活化及促炎细胞因子的产生。
