15d-PGJ_2对神经元氧糖剥夺/复氧损伤的影响

目的观察过氧化物酶体增殖物激活受体γ(PPAR-γ)激动剂15d-PGJ_2对神经元氧糖剥夺/复氧(OGD/R)损伤的影响,并探讨其可能的作用机制。方法采用原代培养的小鼠胎鼠大脑皮质神经元,建立OGD/R损伤模型。将培养的神经元细胞进行分组,Western blot检测各组神经元内LC3-1、LC3-Ⅱ、Bcl-2、Beclin1、AMPK、m TOR及p70S6K等蛋白的表达;MTT法检测细胞活性,乳酸脱氢酶(LDH)漏出率测定细胞毒性。结果 OGD/R损伤6 h、12 h及24 h后神经元LC3-Ⅱ、Beclin 1表达明显增高,而p62表达持续下降,神经元的生存率明显下降,LDH漏出率增加。15d-PGJ_2可显著降低LC3-Ⅱ和Beclin 1表达水平,改善神经元的生存率及降低LDH的漏出率。OGD/R后Bcl-2的蛋白表达水平均明显减少,而Beclin 1表达则显着增加。15d-PGJ_2预处理显著增加OGD/R 24 h的Bcl-2表达量。利用Bcl-2 siRNA或scRNA转染神经元细胞发现,Bcl-2 siRNA可消除15d-PGJ_2对OGD/R后各时间点的抑制效应。结论 15d-PGJ_2能够有效地对神经元OGD/R损伤起到保护作用,其机制可能是通过上调Bcl-2的表达进而抑制自噬的发生实现的。

Effect of 15d-PGJ2 on the neuron oxygen-glucose deprivation/reperfusion injury

Objective To investigate the effect of PPAR-γagonist 15d-PGJ2 on the neuron oxygen-glucose depriva-tion/reperfusion injury and to clarify the underlying mechanism .Methods In this study,We established the cerebral ische-mia-reperfusion models in vitro .Primary cortical neuron was exposed to a paradigm of ischemic insult by using an oxygen -glucose deprivation/reperfusion ( OGD/R ) device and were randomly divided into groups according to the experimental scheme.The expression of LC3、Bcl-2、Beclin1、AMPK、mTOR and p70S6K protein were deterined by Western blot .The neuronal cell viability was detected by MTT and the cytotoxicity was determined by lactate dehydrogenase (LDH) leakage. Results The expression of LC3-Ⅱand Beclin 1 protein significantly increased ,while the p62 protein expression decreased obviously .The neuronal cell viability was decreased and the LDH leakage was increased evidently .The LC3-Ⅱand Beclin 1 protein expression were decreased significantly by the treatment of 15d-PGJ2 ,the neuronal cell viability was improved and the LDH leakage was decreased .The Bcl-2 protein expression decreased while the Beclin 1 protein expression increased ob-viously in the OGD/R injury model.And the pretreatment of 15d-PGJ2 could upregulate Bcl-2 protein expression.Bcl-2 siR-NA could abrogate the effect of 15d-PGJ2 on OGD/R injury model.Conclusions Our study demonstrated that 15d-PGJ2 could effectively protect the neuronal OGD/R injury,the underlying mechanism may involve in the upregulation of Bcl-2 protein expression and inhibit the process of autophagy .