周经霞;陈琳;陈擘璨;邢槐杰;闫丽敏;曾超胜;吴海荣;黄裕盛;陈接桂;

• 1:海南医学院第二附属医院神经内科

摘要(Abstract)：

目的探讨血管内皮生长因子(VEGF)对氧糖剥离再灌注(OGD/R)诱导的大鼠原代神经元损伤的保护作用及机制。方法培养SD大鼠原代神经元,并将细胞随机分为对照组、OGD/R组以及VEGF 25、50、100 ng/mL组,采用OGD/R诱导细胞损伤。采用CCK8检测细胞活性,采用Hoechst染色检测细胞凋亡,酶联免疫法检测各组细胞培养液乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)和丙二醛(MDA)水平;采用免疫印迹法检测各组pro-Caspase-3、Bax、Bcl-2、cleaved Caspase-3、核因子相关因子2(Nrf2)、醌氧化还原酶(NQO1)、血红素氧合酶-1(HO-1)、磷酸化核因子κB抑制因子α(p-IκBα)及核因子-κB(NF-κB)P65的表达水平;利用siRNA沉默Nrf2,采用RT-PCR检测Nrf2和NF-κB P65的表达。结果与对照组比较,OGD/R组细胞活性、SOD水平、GSH-Px水平以及Nrf2、NQO1和HO-1蛋白表达水平均明显降低(P<0.05或P<0.01),细胞凋亡率、Bax/Bcl-2比值、cleaved-Caspase-3/pro-Caspase-3、LDH、MDA表达水平以及p-IκBα和NF-κB p65表达水平明显升高(P<0.05或P<0.01);与OGD/R组比较,VEGF 25、50、100 ng/mL组细胞活性以及SOD和GSH-Px表达水平明显升高,而细胞凋亡率、Bax/Bcl-2比值、pro-Caspase-3/cleaved Caspase-3比值以及LDH和MDA表达水平显著降低,Nrf2、NQO1和HO-1蛋白表达显著上调,p-IκBα和NF-κB p65表达明显下调(均P<0.05);此外,si-Nrf2能明显减弱VEGF (100 ng/mL)对Nrf2和NF-κB p65表达的调控作用,减弱VEGF(100 ng/mL)对细胞活性的促进作用及细胞凋亡的抑制作用(P<0.05或P<0.01)。结论 VEGF能通过调控Nrf2/HO-1和NF-κB通路减轻OGD/R诱导大鼠原代神经元损伤。

关键词(KeyWords)： 再灌注损伤;细胞,培养的;神经元;细胞低氧;细胞凋亡;内皮生长因子;氧化性应激

Abstract:

Keywords:

基金项目(Foundation): 海南省卫生计生行业科研项目(1801032054A2011)

作者(Author): 周经霞;陈琳;陈擘璨;邢槐杰;闫丽敏;曾超胜;吴海荣;黄裕盛;陈接桂;

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