过氧化物酶体增殖物激活受体-γ共同激活因子-1α保护缺血性神经元及其机制探讨

发布时间：2019-03-25 14:48

【摘要】：目的：研究过氧化物酶体增殖物激活受体-γ共同激活因子-lα(Peroxisomeproliferative activated receptor-γ activation of auxiliary factor1Alpha PGC-1α)对氧糖剥夺神经元的保护作用及可能机制探讨。方法：本研究采用原代胚胎小鼠大脑皮层神经元作为载体，氧-糖剥夺作为缺血性脑卒中的体外模拟方法，细胞随机分为五组：正常组，氧-糖剥夺（OGD）组；OGD+PGC-1α过表达质粒组（过表达质粒组）；OGD+PGC-1α iRNA组（干扰质粒组），OGD+空载质粒（空载质粒组）。各组在氧糖剥夺24小时后，噻唑蓝比色分析法(MTT)检测神经细胞存活率。流式细胞仪检测神经细胞死亡率及氧自由基(ROS)量，以采用蛋白印迹技术检测PGC-1α、解偶联蛋白-2（UncouplingProtein-2,UCP2）的表达，，另外，利用质粒干扰技术抑制PGC-1α的表达和过表达。结果：OGD组较正常组神经元存活率明显下降（P＜0.01）；予以PGC-1α过表达质粒预处理后，PGC-1α蛋白水平明显上调，其神经元存活率升高，与OGD组及空载质粒组比较具有显著差异性（P＜0.01），而PGC-1αiRNA质粒预处理后神经元存活率明显降低。OGD组神经元死亡率明显上升，PGC-1α过表达质粒组细胞死亡率下降，与OGD组和空载质粒组比较具有统计学差异(P＜0.01)，干扰质粒组细胞死亡率明显升高与PGC-1α过表达组比较有统计学意义（P＜0.01）。OGD组神经元氧自由基（ROS）含量明显高于正常组升高（P＜0.01)；过表达质粒组ROS水平降低，与OGD组及空载质粒组ROS含量比较其差异具有统计学意义(P＜0.01)，干扰质粒组ROS含量明显升高。研究的结果还显示过表达组UCP-2表达较OGD组显著上调，经PGC-1α干扰质粒预处理后，UCP-2表达明显被下调。结论：PGC-α具有抑制氧糖剥夺神经毒性作用；PGC-1α可能通过上调线粒体UCP-2表达，抑制氧自由基生成，发挥神经保护作用。
[Abstract]:Objective: To study the protective effect and possible mechanism of peroxisome proliferator-activated receptor-activated receptor-activation of auxin factor1Alpha PGC-1. Methods: The primary embryonic mouse cerebral cortex neurons were used as the carrier, and the oxygen-sugar deprivation was used as an in-vitro simulation method of the ischemic stroke. The cells were randomly divided into five groups: normal group, oxygen-sugar deprivation (OGD) group, OGD + PGC-1 and overexpressing plasmid group (over-expression plasmid group). ); OGD + PGC-1, iRNA group (interference plasmid group), OGD + no-load plasmid (no-load plasmid group) ). After a 24-hour deprivation of oxygen for each group, the cell survival was detected by a blue-blue colorimetric assay (MTT). The expression of PGC-1 and the expression of unconjugated protein-2 (UCP2) were detected by flow cytometry. The expression of PGC-1 and the overexpression of PGC-1 were also inhibited by plasmid-interference technique. The results showed that the survival rate of the neurons in the OGD group was significantly lower than that in the normal group (P <0.01); after the pre-treatment of the PGC-1 expression plasmid, the level of the PGC-1 protein was significantly increased, and the survival rate of the neurons was increased, and there was a significant difference with the OGD group and the no-load plasmid group (P <0. 01), and the survival of neurons after pre-treatment of PGC-1-iRNA plasmid In the OGD group, the death rate of the cells increased significantly, and the cell death rate of the PGC-1-overexpressing plasmid group decreased significantly (P <0.01), and the cell death rate of the interfering plasmid group was significantly higher than that of the PGC-1-overexpressing group (P <0.01). The content of ROS in the OGD group was significantly higher than that in the normal group (P <0.01). The ROS level of the overexpressing plasmid group was decreased, and the content of ROS in the OGD group and the no-load plasmid group was statistically significant (P <0.01), and the ROS content of the interfering plasmid group was bright. The results of the study also showed that the expression of UCP-2 in the overexpressing group was up-regulated by the OGD group, and the expression of UCP-2 was obvious after the pretreatment of the PGC-1 interference plasmid. Conclusion: PGC-1 can inhibit the neurotoxicity of oxygen. PGC-1 may play a role in inhibiting the formation of oxygen free radicals by up-regulating the expression of UCP-2 in mitochondria and inhibiting the formation of oxygen free radicals.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R459.7

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