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目的确定羊种布鲁杆菌外膜蛋白31(Omp31)对BV-2小胶质细胞自噬形成的影响。方法 (0.17、0.50、1.50、4.50、13.50)μg/m L Omp31分别处理BV-2细胞6 h,实时定量PCR检测微管相关蛋白1轻链3B(LC3B)mRNA的表达,Western blot法检测LC3BⅡ蛋白、LC3B蛋白的表达;采用透射电镜技术观察各组细胞自噬体,ELISA检测培养细胞上清液肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、IL-10的水平。结果 Omp31处理增加BV-2小胶质细胞LC3BⅡ蛋白水平,且0.5μg/m L Omp31处理LC3BⅡ蛋白增加最明显;0.5μg/m L以下的Omp31对LC3B mRNA的表达有促进作用,大于0.5μg/m L Omp31则抑制LC3B mRNA的表达;0.5μg/m L Omp31能诱导BV-2细胞形成较多自噬体;Omp31能诱导BV-2细胞表达TNF-α、IL-6,但抑制IL-10的表达。结论 0.5μg/m L剂量的Omp31能诱导BV-2细胞发生明显自噬。
Objective To determine the effect of Omp31 on the autophagy of BV-2 microglia. Methods BV-2 cells were treated with Omp31 for 6 h respectively. The expression of LC3B mRNA was detected by real-time quantitative PCR. The expression of LC3BⅡ The expression of TNF-α, IL-6 and IL-10 in supernatant of culture supernatant was detected by transmission electron microscopy. Level. Results Omp31 treatment increased the level of LC3BⅡ protein in BV-2 microglial cells, and the most significant increase of LC3BⅡ protein was observed in 0.5μg / mL L Omp31 treatment. Omp31 at 0.5μg / mL reduced the expression of LC3B mRNA more than 0.5μg / Omp31 induced the expression of TNF-α and IL-6 in BV-2 cells, but inhibited the expression of IL-10 expression. Conclusion Omp31 at a dosage of 0.5μg / m L can induce significant autophagy in BV-2 cells.