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目的:探讨线粒体可溶蛋白诱导小胶质细胞活化作用及和厚朴酚对小胶质细胞活化的影响。方法:培养BV2小胶质细胞,分空白对照组(control)、线粒体可溶蛋白组(MDP)、和厚朴酚干预组(HNK)。ELISA检测不同浓度线粒体可溶蛋白(MSP)刺激不同时间的细胞上清液中IL-6和TNF-α含量;RT-PCR半定量法测定各组转录因子Klf4的表达情况;倒置相差显微镜观察各组细胞形态变化。结果:1.ELISA:MSP(10μg/ml,100μg/ml)处理7h时,IL-6和TNF-α含量与control组比较显著升高(P<0.05);100μg/ml MSP处理1、3、5、7h时,IL-6和TNF-α表达与control组比较明显升高(P<0.05),且7h时HNK组含量明显低于MDP组(P<0.05)。2.RT-PCR结果显示MDP组KLP4的表达显著高于空白对照组和HNK组(P<0.05)。3.倒置相差显微镜观察示空白组胶质细胞形态呈静息状态;MDP组小胶质细胞的胞体变圆或者椭圆,突起消失,呈“阿米巴”状;HNK组活化状态的小胶质细胞明显减少。结论:线粒体可溶蛋白可以激活小胶质细胞,促进白细胞介素IL-6和TNF-α的释放,和厚朴酚能够有效地抑制线粒体可溶蛋白诱导的小胶质细胞活化,其机制可能是通过下调Klf4的表达发挥作用。
Objective: To investigate the mitochondrial soluble protein-induced microglial activation and honokiol on microglial activation. Methods: BV2 microglial cells were cultured and divided into blank control group, mitochondrial soluble protein group (MDP) and honokiol intervention group (HNK). The contents of IL-6 and TNF-α in supernatant of different concentrations of mitochondrial soluble protein (MSP) were measured by ELISA. The expression of Klf4 in each group was determined by reverse transcription-polymerase chain reaction (RT-PCR) Group cell morphology changes. The levels of IL-6 and TNF-α in ELISA group were significantly increased compared with control group (P <0.05) after treatment with ELISA (10μg / ml, 100μg / ml) At 5 and 7h, the expression of IL-6 and TNF-αwere significantly increased compared with control group (P <0.05), and the content of HNK group was significantly lower than that of MDP group at 7h (P <0.05). The results of RT-PCR showed that the expression of KLP4 in MDP group was significantly higher than that in blank control group and HNK group (P <0.05). In inverted phase contrast microscope, the morphology of blank glial cells showed a resting state. The microglial cells in MDP group became round or oval in shape with disappearance of protuberances, showing “amoeba” -like. The activation state of HNK group was small Glial cells decreased significantly. CONCLUSION: Mitochondria soluble protein can activate microglial cells and promote the release of IL-6 and TNF-α. Honokiol can effectively inhibit mitochondrial soluble protein-induced microglial activation, and its mechanism may be Is through down-regulation of Klf4 expression.