探讨BV2细胞活化中是否存在p38MAPK及JAK2-STAT通路的磷酸化激活

Investigate whether p38MAPK and JAK2-STAT pathways can be phosphorylated activation when BV2 cells are activated

目的:探索CD200R在LPS诱导的小胶质细胞炎症模型中的作用以及是否有pho-p38MAPK及pho JAK2-STAT通路的激活。方法:采用小胶质细胞进行研究,将细胞用CD200R抗体及LPS处理,ELISA检测TNF-α及IL-1β的分泌。Western blot检验p38MAPK、JAK2、pho-p38MAPK及pho-JAK2-STAT表达。为证实这两条通路是炎症的下游通路,分别应用两者的阻断剂处理细胞后再次检测炎症因子的分泌。结果:在小胶质细胞表面有CD200R的表达;应用CD200R的阻断性抗体后,TNF-α及IL-1β分泌均增多,较对照组及LPS组比较差异具有统计学意义(P<0.05),并且有pho-p38MAPK及pho-JAK2-STAT的激活;阻断剂SB203580及AG490均能抑制TNF-α及IL-1β的分泌。结论:pho-p38MAPK及pho-JAK2-STAT两条通路是LPS及CD200R抗体诱导的小胶质细胞炎症反应的下游通路。

Objective： To explore the important role of CD200 R in the microglia inflammation model induced by LPS as well as whether there is the activation of the pho-p38 MAPK and pho-JAK2-STAT pathway or not. Methods： Microglia was used for this study.Immunofluorescence confirms there was the expression of CD200 R in microglia. Enzyme-linked immunosorbent assay detected the secretory content of TNF-α and IL-1β after using CD200R-blocking antibody and LPS handle the microglia. Western blot disclose whether p38 MAPK and JAK2-STAT pathway can be phosphorylated or not. Finally,we detect the secretion of TNF-α,IL-1β after applicating different doses of channel blockers. Results： We observed that CD200 R expresses on the surface of microglia. To determine the relationship of CD200 R and inflammatory cytokines,we tested whether blocking CD200 R promotes the release of inflammatory cytokines or not. After using administrating antibodies of CD200 R,TNF-α and IL-1β secreted by microglia was significantly increased and phop38 MAPK and pho-JAK2-STAT were activated,which let us predict that pho-p38 MAPK and pho JAK2-STAT participate in the microglia activation of LPS induction. To further confirm the pho-p38 MAPK and pho-JAK2-STAT pathway were involved in microglia activation,we apply SB203580 and AG490 to inhibit both pathways respectively,and then we found that the number of TNF-α and IL-1β was significantly decreased,and the number of TNF-α and IL-1β was negatively correlation with the concentration of AG490 and SB203580. Conclusion： Not only pho-p38 MAPK but also pho-JAK2-STAT are involved in microglia activation induced by LPS and CD200 R antibody response.