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[小胶质细胞中PRDX6的敲低降低了OGD/R损伤后神经元的活力]

[Knockdown of PRDX6 in microglia reduces neuron viability after OGD/R injury].

作者信息

Tan Li, Zhao Yong, Jiang Beibei, Yang Bo, Zhang Hui

机构信息

Department of Pathology, Molecular Medicine and Cancer Research Center, College of Basic Medical Sciences, Chongqing Medical University, Chongqing 400016, China.

Department of Pathology, Molecular Medicine and Cancer Research Center, College of Basic Medical Sciences, Chongqing Medical University, Chongqing 400016, China. *Corresponding author, E-mail:

出版信息

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2016 Aug;32(8):1014-20.

Abstract

Objective To observe the effects of peroxiredoxin 6 (PRDX6) knockdown in the microglia on neuron viability after oxygen-glucose deprivation and reoxygenation (OGD/R). Methods Microglia was treated with lentivirus PRDX6-siRNA and Ca(2+)-independent phospholipase A2 (iPLA2) inhibitor, 1-hexadecyl-3-(trifluoroethgl)-sn-glycerol-2 phosphomethanol (MJ33). Twenty-four hours later, it was co-cultured with primary neuron to establish the microglia-neuron co-culture OGD/R model. According to the different treatment of microglia, the cells were divided into normal group, OGD/R group, negative control-siRNA treated OGD/R group, PRDX6-siRNA treated OGD/R group and PRDX6-siRNA combined with MJ33 treated OGD/R group. Western blot analysis and real-time quantitative PCR were respectively performed to detect PRDX6 protein and mRNA levels after knockdown of PRDX6 in microglia. The iPLA2 activity was measured by ELISA. MTS and lactate dehydrogenase (LDH) assay were used to measure neuron viability and cell damage. The oxidative stress level of neuron was determined by measuring superoxide dismutase (SOD) and malonaldehyde (MDA) content. Results In PRDX6-siRNA group, neuron viability was inhibited and oxidative stress damage was aggravated compared with OGD/R group. In PRDX6-siRNA combined with MJ33 group, cell viability was promoted and oxidative stress damage was alleviated compared with PRDX6-siRNA group. Conclusion PRDX6 in microglia protects neuron against OGD/R-induced injury, and iPLA2 activity has an effect on PRDX6.

摘要

目的 观察小胶质细胞中过氧化物还原酶6(PRDX6)基因敲低对氧糖剥夺复氧(OGD/R)后神经元活力的影响。方法 用慢病毒PRDX6-siRNA和钙离子非依赖性磷脂酶A2(iPLA2)抑制剂1-十六烷基-3-(三氟乙基)-sn-甘油-2-磷酸甲醇(MJ33)处理小胶质细胞。24小时后,将其与原代神经元共培养,建立小胶质细胞-神经元共培养OGD/R模型。根据小胶质细胞的不同处理方式,将细胞分为正常组、OGD/R组、阴性对照-siRNA处理的OGD/R组、PRDX6-siRNA处理的OGD/R组和PRDX6-siRNA联合MJ33处理的OGD/R组。分别采用蛋白质免疫印迹分析和实时定量PCR检测小胶质细胞中PRDX6基因敲低后的PRDX6蛋白和mRNA水平。用酶联免疫吸附测定法(ELISA)检测iPLA2活性。采用噻唑蓝(MTS)法和乳酸脱氢酶(LDH)测定法检测神经元活力和细胞损伤情况。通过检测超氧化物歧化酶(SOD)和丙二醛(MDA)含量来测定神经元的氧化应激水平。结果 与OGD/R组相比,PRDX6-siRNA组神经元活力受到抑制,氧化应激损伤加重。与PRDX6-siRNA组相比,PRDX6-siRNA联合MJ33组细胞活力得到促进,氧化应激损伤减轻。结论 小胶质细胞中的PRDX6可保护神经元免受OGD/R诱导的损伤,且iPLA2活性对PRDX6有影响。

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