ATG12基因对易卒中自发性高血压大鼠大脑组织自噬的影响  

Effect of ATG12 gene on autophagy in brain tissue of stroke-prone spontaneously hypertensive rats

作　　者：李浩东[1] 徐虹[1] 范永新[1] 刘猛[1] 朱月敏[1] 马俊平[1] 栾松[1] LI Haodong;XU Hong;FAN Yongxin;LIU Meng;ZHU Yuemin;MA Junping;LUAN Song(Department of Neurology,Zhuozhou City Hospital,Zhuozhou 072750,China)

机构地区：[1]涿州市医院神经内科

出　　处：《山西医科大学学报》2019年第11期1556-1560,共5页Journal of Shanxi Medical University

基　　金：保定市科技计划项目(17ZF233)

摘　　要：目的探讨自噬相关蛋白12(autophagy-related protein 12,ATG12)对易卒中自发性高血压大鼠(stroke-prone spontaneously hypertensive rats,SHR/SP)大脑组织自噬的影响。方法将30只SHR/SP分为SHR/SP模型(SHR/SP)组、对照病毒(NC shRNA SHR/SP)组和ATG低表达病毒(ATG12 shRNA SHR/SP)组,其中ATG12 shRNA SHR/SP和NC shRNA SHR/SP组大鼠分别为通过尾静脉注射病毒构建ATG12低表达和对照大鼠,10只WKY(Wistar-Kyoto)大鼠作为SHR/SP的空白对照(WKY组)。荧光定量PCR检测WKY和SHR/SP大脑组织中ATG12 mRNA表达水平。通过大鼠血压计检测各组大鼠血压,分光光度法检测各组大鼠大脑组织氧化应激指标SOD和MDA水平;Western blot检测各组大鼠大脑组织自噬相关蛋白LC3A/B、ATG12和P62蛋白表达水平。结果与WKY组大鼠相比,SHR/SP模型组大鼠大脑组织ATG12 mRNA水平显著升高(P <0. 01)。血压检测结果显示,与WKY大鼠组比较,SHR/SP组大鼠血压显著升高,与NC shRNA SHR/SP组比较,ATG12 shRNA SHR/SP组大鼠血压显著降低(P <0. 01)。与WKY组比较,SHR/SP模型组大鼠大脑组织SOD活性显著降低,MDA含量显著升高(P <0. 01);与NC shRNA SHR/SP组比较,ATG12 shRNA-SHR/SP组大鼠大脑组织SOD活性显著升高,MDA含量显著降低(P <0. 01)。Western blot结果显示,与WKY组比较,SHR/SP模型组大鼠大脑组织ATG12和LC3A/B蛋白表达显著升高,P62蛋白表达显著降低;与NC shRNA SHR/SP组比较,ATG12 shRNA-SHR/SP组大鼠大脑组织ATG12和LC3A/B蛋白表达显著降低,P62蛋白表达显著升高(P <0. 01)。结论易卒中自发性高血压大鼠大脑组织ATG12高表达,抑制ATG12表达能够抑制大鼠脑组织自噬水平,降低大鼠血压,可能与抑制大鼠脑组织氧化应激水平有关。Objective To investigate the effect of autophagy-related protein 12( ATG12) on autophagy in brain tissue of stroke-prone spontaneously hypertensive rats( SHR/SP). Methods The ATG12 low expression rat model was constructed by injecting adeno-associated virus( AAV) into the tail vein. Thirty rats were divided into SHR/SP model group,negative control virus SHR/SP model( NC shRNA SHR/SP) group and ATG low expression virus SHR/SP model( ATG12 shRNA-SHR/SP) group. The ATG12 low expression( ATG12 shRNA-SHR/SP) and control( NC shRNA SHR/SP) rats were injected with adeno-associated virus( AAV) into the tail vein.Ten WKY rats were chosen as blank controls( WKY group). The expression level of ATG12 mRNA in brain tissue was detected by real-time PCR. The blood pressure was measured. The levels of SOD and MDA in brain tissues were detected by spectrophotometry. The expression levels of autophagy-related proteins LC3 A/B,ATG12 and P62 in brain tissues were detected by Western blot. Results Compared with WKY group,the ATG12 mRNA level in the brain was significantly increased in SHR/SP model group( P < 0. 01). The blood pressure in SHR/SP group was significantly higher than that in WKY group. Compared with NC shRNA SHR/SP group,the blood pressure was significantly decreased in ATG12 shRNA SHR/SP group( P < 0. 01). Compared with WKY group,the SOD activity in the brain tissue was significantly decreased in SHR/SP model group,while the MDA content was significantly increased( P < 0. 01).Compared with NC shRNA SHR/SP group,the SOD activity in the brain tissue was significantly increased in ATG12 shRNA-SHR/SP group,while the MDA content was significantly decreased( P < 0. 01). Western blot results showed that compared with WKY group,the expression of ATG12 and LC3 A/B protein in the brain was significantly increased in SHR/SP model group,while the expression of P62 protein was significantly decreased. Compared with NC shRNA SHR/SP group,the expression of ATG12 and LC3 A/B protein in brain tissue was significantly decreased in ATG12 s

关 键 词：ATG12 高血压 脑梗死 氧化应激 细胞自噬 

分 类 号：R743[医药卫生—神经病学与精神病学]