PARK7通过调控Trx2对缺糖缺氧诱导的大鼠皮质神经元Necroptosis的保护机制  

PARK7 protects rat cortical neurons from Necroptosis induced by oxygen-glucose deprivation in primary cortical neurons via Trx2 pathway

作　　者：陈巍巍[1] 张亚洁 刘芷含 程萍 黄文娟[1] 谢志远[1] 张侠[1] CHEN Weiwei;ZHANG Yajie;LIU Zhihan;CHENG Ping;HUANG Wenjuan;XIE Zhiyuan;ZHANG Xia(Xuzhou Central Hospital, Xuzhou 221009, Jiangsu, China;Xuzhou Medical University, Xuzhou 221004, Jiangsu, China;The Affiliated Hospital of Xuzhou Medical University, Xuzhou 221002, Jiangsu, China;Bengbu Medical College, Bengbu 233000, Anhui, China)

机构地区：[1]徐州市中心医院,江苏徐州221009 [2]徐州医科大学,江苏徐州221004 [3]徐州医科大学附属医院,江苏徐州221002 [4]蚌埠医学院,安徽蚌埠233000

出　　处：《现代中西医结合杂志》2021年第28期3088-3094,共7页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：国家自然科学基金资助项目(81501138);徐州市科技项目(KC18038,KC19027,KC20108)。

摘　　要：目的探讨帕金森病蛋白7(PARK7)在缺糖缺氧诱导的大鼠皮质神经元Necroptosis信号通路中调控硫氧还蛋白2(Trx2)对NOD样受体蛋白3(NLRP3)炎性小体介导的炎性反应的抑制作用。方法①实验一:分为正常对照组、缺糖缺氧+Caspase抑制剂组、DNCB组。SD大鼠皮质神经元细胞培养至12 d,缺糖缺氧+Caspase抑制剂组、DNCB组均给予Caspase抑制剂预处理,DNCB同时给予DNCB预处理,缺氧2 h/复氧12 h,Western blot检测线粒体中Trx2蛋白表达情况,全自动生化分析仪检测培养基中乳酸脱氢酶(LDH)水平。②实验二:分为正常对照组、缺糖缺氧+Caspase抑制剂组、慢病毒敲减PARK7组(sh PARK7组)、慢病毒扩增PARK7组(OE-PARK7组),各组给予相应处理后,Western blot检测PARK7、Trx2、NLRP3、IL-1β蛋白表达情况,全自动生化分析仪检测培养基中LDH水平。③实验三:TrxR活性检测分为正常对照组、缺糖缺氧+Caspase抑制剂组、DNCB组、sh PARK7组、OE-PARK7组、sh PARK7+DNCB组、OE-PARK7+DNCB组,各组给予相应处理后检测TrxR活性;神经元受损程度实验分为正常对照组、缺糖缺氧+Caspase抑制剂组、DNCB组、OE-PARK7组和OE-PARK7+DNCB组,各组给予相应处理后,检测缺氧2 h和复氧2 h、6 h、12 h、24 h后LDH水平。结果①DNCB组Trx2表达水平与缺糖缺氧+Caspase抑制剂组比较差异无统计学意义(P>0.05),LDH水平明显高于缺糖缺氧+Caspase抑制剂组(P<0.05)。②OE-PARK7组PARK7、Trx2蛋白相对表达量均明显高于缺糖缺氧+Caspase抑制剂组和sh PARK7组(P均<0.05),sh PARK7组均明显低于缺糖缺氧+Caspase抑制剂组(P均<0.05);OE-PARK7组NLRP3、IL-1β蛋白相对表达量及LDH水平均明显低于缺糖缺氧+Caspase抑制剂组和sh PARK7组(P均<0.05),sh PARK7组均明显高于缺糖缺氧+Caspase抑制剂组(P均<0.05)。③缺糖缺氧+Caspase抑制剂组、sh PARK7组、OE-PARK7组TrxR活性均明显增高于正常对照组(P均<0.05),3组间比较差异均无统计学意义(P均>0.Objective It is to investigate the inhibition of Parkinson’s disease protein 7(PARK7)on inflammatory response mediated by NOD-like receptor protein 3(NLRP3)inflammasome through regulating thioredoxin 2(Trx2)in the Necroptosis signaling pathway of rat cortical neurons induced by hypoglycemia and hypoxia.Methods①ExperimentⅠ:the rats were divided into normal control group,hypoglycemia and hypoxia+Caspase inhibitor group,and DNCB group.The cortical neuron cells of SD rats were cultured for 12 days.Both the hypoglycemia and hypoxia+Caspase inhibitor group and DNCB group were pretreated with Caspase inhibitor,the DNCB group was pretreated with DNCB at the same time.After hypoxia for 2h/reoxygenation for 12h,the expression of Trx2 protein in mitochondria was detected by Western blot,the level of lactate dehydrogenase(LDH)in the culture medium was detected by automatic biochemical analyzer.②ExperimentⅡ:The rats were divided into normal control group,hypoglycemia and hypoxia+Caspase inhibitor group,sh PARK7 group,OE-PARK7 group,after given corresponding treatment in each group,the protein expression of PARK7,Trx2,NLRP3,IL-1βwas detected by Western blot,the level of LDH in the culture medium was detected by automatic biochemical analyzer.③ExperimentⅢ:TrxR activity detection was divided into normal control group,hypoglycemia and hypoxia+Caspase inhibitor group,DNCB group,sh PARK7 group,OE-PARK7 group,sh PARK7+DNCB group,OE-PARK7+DNCB group,after given corresponding treatment in each group,the level of LDH was detected after 2h of hypoxia and 2 h,6 h,12 h,and 24 h of reoxygenation.Results①The expression level of Trx2 in the DNCB group was not significantly different from that in the hypoglycemia and hypoxia+Caspase inhibitor group(P>0.05),and the LDH level was significantly higher than that in the hypoglycemia and hypoxia+Caspase inhibitor group(P<0.05).②The relative expressions of PARK7 and Trx2 proteins in the OE-PARK7 group were significantly higher than those in the hypoglycemia and hypoxia+Caspase inh

关 键 词：Necroptosis信号通路 帕金森病蛋白7 硫氧还蛋白2 NOP样受体蛋白 

分 类 号：R-332[医药卫生]