活化星形胶质细胞中线粒体生物发生增强

摘要

Objective To investigate the changes of mitochondrial biogenesis in activated astrocytes (ASs).Methods The ASs isolated from neonatal rat cerebral cortex were purified and cultured; and then,they were randomly divided into 4 groups:control group,and lipopolysaccharide (LPS) and interferon-γ (IFN-γ) insulted 6,12 and 24 h groups.The levels of tumor necrosis factor-α (TNF-o) and interleukin-6 (IL-6) were detected by enzyme-linked immuno sorbent assay.The expressions of peroxisomal proliferator-activated receptor gamma coactivator 1 alpha (PGC-1α),cytochrome C oxidase 1 (COX1),COX2,optic atrophy 1 gene protein (OPA1) and dynamin-related protein 1 (DRP1) were detected by Western blotting.The mitochondrial DNA relative content was detected by real-time polymerase chain reaction (RT-PCR).The mitochondrial volume density was observed by transmission electron microscopy.Results The levels of TNF-α and IL-6 in the LPS and IFN-γinsulted 6,12 and 24 h groups were significantly increased as compared with those in the control group (P＜0.05).The protein expressions of PGC-1α,COX1,COX2,OPA1 and DRP1 in LPS and IFN-γ insulted 6 and 12 h groups were significantly increased as compared with those in the control group (P＜0.05).The mitochondrial DNA relative contents in LPS and IFN-γ insulted 12 and 24 h groups were significantly increased as compared with those in control group (P＜0.05).The mitochondrial volume density was dramatically increased in LPS and IFN-γ insulted 24 h group as compared with that in the control group (P＜0.05) Conclusion Mitochondrial biogenesis is increased in ASs.%目的 观察星形胶质细胞(AS)活化后其线粒体生物发生的变化情况.方法 分离新生大鼠大脑皮质AS并纯化培养,随机分为正常对照组、脂多糖(LPS)+干扰素-γ(IFN-γ)联合刺激6h组、12h组和24 h组,酶联免疫吸附法检测AS分泌肿瘤坏死因子(TNF-α)和白细胞介素-6(IL-6)浓度,蛋白质印迹法检测过氧化物酶体增殖物激活受体γ辅助激活因子-1α(PGC-1α)、线粒体细胞色素C氧化酶亚单位1(COX1)、线粒体细胞色素C氧化酶亚单位2(COX2)、视神经萎缩1基因蛋白(OPA1)和动力相关蛋白1(DRP1)蛋白的表达情况,实时定量聚合酶链反应(RT-PCR)检测AS线粒体DNA相对含量,透射电镜观察计算AS线粒体体积密度.结果 6h组、12h组和24 h组TNF-α和IL-6浓度均较正常对照组显著升高,差异具有统计学意义(P＜0.05);6h组和12h组PGC-1α、COX1、COX2、OPA1和DRP1蛋白浓度均较正常对照组显著增加,差异具有统计学意义(P＜0.05); 12 h组和24 h组AS线粒体DNA相对含量较正常对照组显著增加,差异具有统计学意义(P＜0.05);24 h组AS线粒体体积密度较正常对照组显著增加,差异具有统计学意义(P＜0.05).结论 线粒体生物发生在活化AS中增强.