丙型肝炎病毒阳性血清对小胶质细胞TLR7蛋白及炎性因子表达的影响  

The influence of hepatitis C virus-positive serum on the expressions of TLR7 protein and other inflammatory factors in microglial cells

作　　者：赵莉[1] 王振海[1] 

机构地区：[1]宁夏医科大学附属医院神经内科宁夏颅脑疾病重点实验室,银川750004

出　　处：《现代免疫学》2011年第1期52-55,共4页Current Immunology

基　　金：国家自然科学基金资助项目(30760076);教育部基金资助项目(Z2008-1-75002);宁夏自然科学基金资助项目(NZ0790)

摘　　要：通过观察丙型肝炎病毒阳性血清处理后小鼠小胶质细胞BV2(BV2细胞)内TLR7蛋白及TNF-α、IFN-α的表达变化,探讨HCV对中枢神经系统损伤的免疫发病机制。用20%HCV阳性血清处理BV2细胞,在处理后24 h收集细胞及培养上清液,应用间接免疫荧光流式细胞术及酶联免疫吸附法(ELISA)分别检测细胞内TLR7蛋白和培养上清液中TNF-α、IFN-α的表达,同时设同型对照、空白组和正常人血清组。结果表明,BV2细胞内有TLR7蛋白的表达,其中HCV阳性血清组表达明显升高,较空白组及正常人血清组比较具有统计学意义(P<0.05),正常人血清组与空白对照组比较无统计学意义。HCV阳性血清处理BV2细胞24h后TNF-α、IFN-α水平均较空白组及正常人血清组增高,具有统计学意义(P<0.05),正常人血清组与空白对照组比较无统计学意义(P>0.05)。本研究初步阐明了HCV阳性血清处理后BV2细胞中TLR7表达显著升高,TLR7激活后能够启动BV2细胞的固有免疫反应,因此TLR7可能在HCV导致的CNS的发病中发挥重要作用。In order to clarify the immune pathogenesis of damages in central nervous system induced by HCV through the observation on the alterations in the expressions of TLR7 protein, TNF-α and IFN-α in murine mieroglial cells treated with HCV-positive serum, the murine microglial cell BV2 cells were cultured and infected with 20% HCV positive-serum. The BV2 cells and the cultural supernatants were collected 24 hrs after infection and indirect immunofluorescence flow cytometry and enzyme-linked immunosorbent assay（ELISA） were used to detect the expressions of TLR7 protein, TNF-α, and IFN-α. Meanwhile, the isotype control, blank control and normal human serum control were set up. It was demonstrated that the expression of TLR7 protein could be detected in BV2 cells, in which the expression of TLR7 protein expression of the HCV-positive serum group was significantly higher than those of the blank control and group of normal human serum control and the difference was significant （P〈0.05）, while no statistical difference existed between the group of blank control and group of normal human serum control （P〉0.05）. Also, TNF-α, IFN-α levels in culture supernatant increased significantly （P〈0.05） than the control group and the normal serum group, normal serum group compared with the control group showed no statistical significance （P〉0.05）. From the results of the present study, it is evident that TLR7 may be activated by HCV positive serum and pro- moted the innate immune response of microglial cells, so it may play a role in the pathogenesis of central nervous system disorders induced by hepatitis C virus.

关 键 词：TLR7 HCV 小胶质细胞 流式细胞术 TNF-α IFN—α 

分 类 号：R74[医药卫生—神经病学与精神病学]