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目的:了解在体外培养条件下丙型肝炎病毒(HCV)刺激小鼠小胶质细胞(BV2)后Toll样受体9(TLR9)蛋白的表达变化及其对MCP-1(人单核细胞趋化蛋白-1)和TNF-α分泌的影响,探讨HCV感染中枢神经系统(CNS)后小胶质细胞TLR9蛋白的表达及其相应细胞因子的变化。方法:体外培养BV2细胞,分为HCV阳性血清组、正常人血清组、空白对照组。每组各20例细胞样本。在12h收集细胞和上清液,用流式细胞术检测TLR9蛋白的表达,ELISA法检测培养上清中MCP-1和TNF-α的含量,并应用SPSS11.0统计软件包进行统计学分析。结果:(1)各组BV2细胞均有TLR9蛋白的表达,HCV阳性血清组中TLR9蛋白的表达高于正常人血清组、空白对照组(P<0.01);正常人血清组和空白对照组无统计学差异(P>0.05);(2)HCV阳性血清组MCP-1和TNF-α含量高于正常人血清组、空白对照组(P<0.01);且HCV阳性血清组TLR9蛋白的表达量与MCP-1和TNF-α的分泌可能呈正相关。结论:HCV阳性血清刺激可使体外培养的BV2细胞表达TLR9蛋白增高,且可能诱导了MCP-1和TNF-α的分泌。TLR9蛋白可能参与了HCV在中枢神经系统感染中的早期固有免疫应答。
OBJECTIVE: To investigate the expression of Toll-like receptor 9 (TLR9) protein in mouse microglia (BV2) stimulated by Hepatitis C virus (HCV) in vitro and its effect on the expression of MCP-1 1) and TNF-α secretion, and to explore the expression of TLR9 protein and the corresponding cytokines in microglia after infection with the central nervous system (CNS). Methods: BV2 cells were cultured in vitro and divided into HCV positive serum group, normal human serum group and blank control group. Each group of 20 cell samples. The cells and supernatant were harvested at 12 hours. The expression of TLR9 protein was detected by flow cytometry. The contents of MCP-1 and TNF-α in culture supernatants were detected by ELISA. The statistical software package SPSS11.0 was used for statistical analysis. Results: (1) The expression of TLR9 protein in BV2 cells of each group was higher than that in normal human serum group and blank control group (P <0.01). The expression of TLR9 protein in HCV positive serum group was higher than that in normal human serum group and blank control group (P> 0.05). (2) The levels of MCP-1 and TNF-α in HCV positive serum were higher than those in normal human serum and blank control group (P <0.01). The expression of TLR9 in HCV positive serum group It may be positively correlated with the secretion of MCP-1 and TNF-α. CONCLUSION: HCV-positive serum can stimulate the expression of TLR9 protein in cultured BV2 cells, and may induce the secretion of MCP-1 and TNF-α. TLR9 protein may be involved in the early innate immune response of HCV in central nervous system infection.