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目的检测HCV刺激小鼠小胶质细胞(BV2细胞)后IL-12、IFN-α分泌变化及其与TLR4的关系。方法用含20%HCV阳性血清的培养液刺激BV2细胞,为HCV血清组,并设置正常血清组和空白对照组。于24 h后用RT-qPCR方法观察各组TLR4相对表达,应用ELISA方法检测各组细胞培养上清液中IL-12、IFN-α的含量,同时观察TLR4阻断后各组IL-12、IFN-α的分泌变化。结果 HCV血清组TLR4相对表达和IL-12、IFN-α分泌均高于正常血清组及空白对照组(P<0.01);阻断TLR4的HCV血清组IL-12、IFN-α分泌明显低于非阻断组(P<0.01),高于正常血清组及空白对照组(P<0.01);正常血清组和空白对照组阻断前后无明显变化(P>0.05)。结论 TLR4可能会通过介导HCV刺激小胶质细胞分泌IL-12、IFN-α等来发挥宿主CNS抗HCV的固有免疫应答。
Objective To detect the changes of IL-12 and IFN-α secretion and the relationship with TLR4 in mice microglia (BV2) stimulated by HCV. Methods BV2 cells were stimulated by culture medium containing 20% HCV-positive serum for HCV serogroups, and normal serum group and blank control group were set up. After 24 h, the relative expression of TLR4 in each group was observed by RT-qPCR method. The contents of IL-12 and IFN-α in the culture supernatants of each group were detected by ELISA. The levels of IL-12, Secretion of IFN-α. Results The relative expression of TLR4 and the secretion of IL-12 and IFN-α in HCV serum were higher than those in normal serum and blank control group (P <0.01). The secretion of IL-12 and IFN-α in HCV serum of TLR4 group was significantly lower than Non-blocking group (P <0.01), higher than normal serum group and blank control group (P <0.01); there was no significant difference between normal serum group and blank control group (P> 0.05). Conclusion TLR4 may play an innate immune response to host CNS against HCV by mediating the secretion of IL-12, IFN-α by microglial cells stimulated by HCV.