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目的观察敲低小胶质细胞中过氧还原素6(PRDX6)对氧糖剥夺再复氧(OGD/R)神经元存活的影响。方法小胶质细胞采用PRDX6-siRNA慢病毒感染,同时培养基中加入非Ca2+依赖性磷脂酶A2(i PLA2)活性抑制剂MJ33,24 h后与原代神经元共培养,并进行OGD/R处理。实验分为正常组、OGD/R组、阴性对照siRNA处理的OGD/R组、PRDX6-siRNA处理的OGD/R组、PRDX6-siRNA联合MJ33处理的OGD/R组。Western blot法检测小胶质细胞PRDX6蛋白的表达,实时定量PCR检测小胶质细胞PRDX6 mRNA的表达。ELISA检测i PLA2活性;MTS、乳酸脱氢酶(LDH)法检测神经元的存活率;采用比色法检测超氧化物歧化酶(SOD)和丙二醛(MDA)含量以反映神经元的氧化应激水平。结果与OGD/R组比较,PRDX6-siRNA处理的神经元存活率降低,MDA增加、SOD降低、氧化应激损伤加重;与PRDX6-siRNA组比较,同时加入i PLA2活性抑制剂MJ33组神经元存活率增加,MDA降低、SOD增加、氧化应激水平下降。结论小胶质细胞PRDX6对神经元氧糖剥夺损伤有保护作用,而i PLA2活性对PRDX6的作用有影响。
Objective To observe the effect of PRDX6 on the survival of neurons in OGD / R after knocking down microglia. Methods The microglia cells were infected with PRDX6-siRNA lentivirus. MJ33, a non-Ca2 + -dependent phospholipase A2 (i PLA2) inhibitor, was added to the culture medium for 24 h, then co-cultured with primary neurons. OGD / R deal with. The experiment was divided into OGD / R group, OGD / R group, OGD / R group, PRDX6-siRNA-treated OGD / R group and PRDX6-siRNA combined with MJ33-treated OGD / R group. The expression of PRDX6 in microglia was detected by Western blot and the expression of PRDX6 mRNA in microglia was detected by real-time quantitative PCR. ELISA was used to detect the activity of PLA2; the survival rate of neurons was detected by MTS and LDH; the contents of superoxide dismutase (SOD) and malondialdehyde (MDA) were measured by colorimetric method to reflect the neuronal oxidation Stress level. Results Compared with OGD / R group, the survival rate of PRDX6-siRNA-treated neurons decreased, MDA increased, SOD decreased, and oxidative stress injury increased. Compared with PRDX6-siRNA group, the neurons survived with addition of i PLA2 inhibitor MJ33 The rate of increase, MDA decreased, SOD increased, the level of oxidative stress decreased. Conclusion Microglia PRDX6 has a protective effect on neuronal oxygen-glucose deprivation injury, while i PLA2 activity has an effect on the effect of PRDX6.