高糖环境对肝星状细胞TLR4 表达及脂多糖诱导的炎症因子表达的影响.docVIP

·386·  doi 10.3969/j.issn.1673-4254.2013.03.16  J South Med Univ, 2013, 33(3): 386-390  基础研究 高糖环境对肝星状细胞 TLR4 表达及脂多糖诱导的炎症因子表 达的影响 史海涛，董 蕾，刘亚萍，贾 淼，赵 刚，赵菊辉，鲁晓岚 西安交通大学医学院第二附属医院，陕西 西安 710004 摘要：目的 研究高糖环境对肝星状细胞 Toll 样受体 4(TLR4)表达及脂多糖（LPS）诱导的炎症因子表达的影响。方法 体外培养 大鼠肝星状细胞株 HSC-T6，采用高糖刺激，Western blot 及 RT-PCR 检测细胞 TLR4 蛋白及 mRNA 表达；低糖或高糖培养基预处 理肝星状细胞 24 h 后采用 LPS（100 ng/ml）刺激，2 h 后 Western blot 检测 NF-κB 核转位，24 h 后 RT-PCR 检测 MCP-1 和 IL-6 mRNA 表达，ELISA 检测细胞上清两种炎症因子的浓度。结果 高糖环境可上调肝星状细胞 TLR4 蛋白及 mRNA 表达（均为 P 0.01），且呈明显的时效-量效关系。高糖可促进肝星状细胞 NF-κB 核转位（P0.01），并促进 MCP-1 和 IL-6 表达及分泌（均为 P 0.01）。高糖预处理可增强 LPS 诱导的肝星状细胞 NF-κB 核转位（P0.01）、MCP-1 和 IL-6 表达及分泌（均为 P0.01）。结论 高 糖可上调肝星状细胞 TLR4 的表达，并可促进脂多糖激活 NF-κB 而增加炎症因子表达及分泌。 关键词：高糖；肝星状细胞；Toll 样受体 4  Effect of high glucose on toll-like  receptor  4  expression and  LPS-induced proinflammatory cytokine production in hepatic stellate cells in vitro 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, Xian 710004, China Abstract: Objective 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. Methods 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. Results High glucose significantly increased the mRNA and protein expressions of TLR4 (P0.01) in a time- and dose-dependent manner. High glucose promoted NF-κB nuclea