聚核糖性死亡在低氧复合丙泊酚致PC12细胞损伤中的作用  

Role of parthanatos in PC12 cell injury induced by propofol combined with hypoxic conditions

作　　者：王维萍 高雯 陈杭 刘玲[1] 涂生芬[1] 杨飞[1] WANG Weiping;GAO Wen;CHEN Hang;LIU Ling;TU Shengfen;YANG Fei(Department of Anesthesiology,National Clinical Research Center of Child Health and Disease,Key Laboratory of Child Developmental Diseases of Ministry of Education,Chongqing Key Laboratory of Pediatrics,Children’s Hospital of Chongqing Medical University,Chongqing,400014,China)

机构地区：[1]重庆医科大学附属儿童医院麻醉科,国家儿童健康与疾病临床医学研究中心,儿童发育疾病研究教育部重点实验室,儿科学重庆市重点实验室,重庆400014

出　　处：《陆军军医大学学报》2022年第10期1025-1032,共8页Journal of Army Medical University

基　　金：国家自然科学基金青年科学基金(31200853)。

摘　　要：目的探讨聚核糖性死亡在低氧环境下丙泊酚致PC12细胞损伤中的作用。方法采用经神经生长因子(nerve growth factor,NGF)诱导分化后的PC12细胞模型,用随机数字表法将细胞分为对照(NC)组(正常培养)、脂肪乳空气对照(CA)组、脂肪乳低氧对照(CH)组、丙泊酚空气(PA)组、丙泊酚低氧(PH)组、3-AB(PARP-1抑制剂)组。采用乳酸脱氢酶(Lactate dehydrogenase,LDH)试剂盒检测上清液中LDH的水平;流式细胞技术检测细胞凋亡;RT-qPCR法检测多聚ADP核糖聚合酶1(poly ADP-ribose polymerase 1,PARP-1)、细胞凋亡诱导因子(apoptosis-inducing factor,AIF)蛋白的mRNA表达水平;Western blot检测PARP-1、多聚ADP核糖(polymerized ADP-ribose,PAR)及AIF蛋白表达水平;细胞免疫荧光技术检测AIF核质分布。结果与NC、CA组比较,PA、PH组LDH水平及细胞凋亡增多(P<0.01),PARP-1和AIF的mRNA和蛋白表达均升高(P<0.05),胞质PAR聚合物形成增多(P<0.01),AIF核迁移显著增加(P<0.01);与CH、PA组比较,PH组LDH水平及细胞凋亡增多(P<0.01),PARP-1 mRNA和蛋白表达均升高(P<0.01),胞质PAR聚合物形成增多(P<0.01),AIF核迁移增加(P<0.01);使用PARP-1抑制剂3-AB预处理可降低低氧复合丙泊酚引起的LDH水平及细胞凋亡增加(P<0.01),同时抑制PARP-1、AIF和PAR蛋白表达上调(P<0.01)。结论低氧环境下丙泊酚可导致PC12细胞发生聚核糖性死亡,抑制聚核糖性死亡可缓解低氧复合丙泊酚对PC12细胞的损伤。Objective To explore the role of parthanatos in propofol-induced injury in PC12 cells under hypoxic conditions.Methods PC12 cells were induced to differentiate with mouse nerve growth factor(NGF),and randomly divided into normal control(NC)group,lipid emulsion solvent+room air(CA)group,lipid emulsion solvent+hypoxia control(CH)group,propofol+room air(PA)group,propofol+hypoxia(PH)group,and 3-AB(PARP-1 inhibitor)group.Lactate dehydrogenase(LDH)kit and flow cytometry were adopted to detect the LDH level in the supernatant and the cell apoptosis,respectively.The mRNA levels of poly ADP-ribose polymerase 1(PARP-1)and apoptosis-inducing factor(AIF)were determined by RT-qPCR,and the protein levels of AIF,PARP-1 and polymerized ADP-ribose(PAR)in each group were measured by Western blotting.The nuclear distribution of AIF in PC12 cells was observed by immunofluorescence assay.Results As compared with the NC and CA groups,the levels of LDH and apoptosis were significantly enhanced in the PA and PH groups(P<0.01);the mRNA and protein levels of PARP-1 and AIF(P<0.05)were increased;and the accumulation of cytoplasmic PAR(P<0.01)and AIF nuclear translocation(P<0.01)were observed as well.In comparison with the CH and PA groups,the levels of LDH and cell apoptosis(P<0.01),the mRNA and protein expression of PARP-1(P<0.01),the formation of cytoplasmic PAR(P<0.01),and the AIF nuclear translocation(P<0.01)were all augmented in the PH group.Pretreatment with PARP-1 inhibitor 3-AB notably reversed the increase in LDH level and cell apoptosis induced by hypoxia combined with propofol(P<0.01),and restrained the up-regulation of PARP-1,AIF and PAR(P<0.01).Conclusion Propofol causes parthanatos of PC12 cells under hypoxic conditions,and inhibition of parthanatos can alleviate the damage in PC12 cells induced by propofol combined with hypoxia.

关 键 词：低氧 丙泊酚 大鼠肾上腺嗜铬瘤细胞 聚核糖性死亡 

分 类 号：R361.3[医药卫生—病理学] R364.4[医药卫生—基础医学]