摘要
目的探讨氧化应激在心脏骤(CA)停引起的小鼠自噬体清除损伤中的作用。方法将30只昆明小鼠随机均分为3组:假手术组、CA组和MnTMPyP组,后两组采用改良的窒息法建立心脏骤停-自主循环恢复(CA-ROSC)小鼠模型。假手术组不进行CA,其余同CA组。MnTMPyP组在建模之前给予一次性腹腔注射6 mg/kg的起氧化物歧化酶(SOD)模拟物MnTMPyP[作为活性氧(ROS)的清除剂]。CA-ROSC后6 h,Western Blot检测各组小鼠大脑皮质自噬体及溶酶体相关指标,试剂盒法检测氧化应激相关指标[ROS、超氧化歧化酶(SOD)和谷胱甘肽过氧化酶(GSH-Px)],TUNEL染色检测细胞死亡情况。结果CA组自噬相关蛋白LC3-Ⅱ/LC3-Ⅰ比值、Beclin-1、泛素(ubiquitin)和SQSTM1/p62表达均显著高于假手术组(P<0.05),MnTMPyP组显著低于CA组(P<0.05)。CA组ROS含量及TUNEL阳性细胞数均显著高于假手术组(P<0.05),MnTMPyP组显著低于CA组(P<0.05)。CA组SOD、GSH-Px、溶酶体关联膜蛋白2(LAMP2)和组织蛋白酶D(Cathepsin D)水平均显著低于假手术组(P<0.05),MnTMPyP组显著高于CA组(P<0.05)。结论氧化应激抑制可缓解CA-ROSC导致的自噬体清除受损及溶酶体功能障碍,抑制神经元死亡。
Objective To investigate the role of oxidative stress in autophagosome clearance injury in mice caused by cardiac arrest(CA).Methods 30 Kunming mice were randomly divided into 3 groups,including sham group,CA group and MnTMPyP group.The mice in latter two groups were given a modified asphyxia method to establish cardiac arrest-restoration of spontaneous circulation(CA-ROSC)mouse model.The mice in sham group did not perform CA,and the rest were the same as the CA group.The mice in MnTMPyP group were given a one-time intraperitoneal injection of 6 mg/kg of the SOD mimetic MnTMPyP(as ROS scavenger)prior to modeling.Western Blot was used to detect autophagosome and lysosome related proteins in cerebral cortex of mice in each group.The oxidative stress related indicators were detected by kit method,and cell death was detected by TUNEL staining.Results The ratio of LC3-II/LC3-I and expressions of Beclin-1,ubiquitin and SQSTM1/p62 in the CA group were significantly higher than those in the sham group(P<0.05),and the MnTMPyP group was significantly lower than the CA group(P<0.05).The content of ROS and numbers of TUNEL positive cells in CA group were significantly higher than those in sham group(P<0.05),and MnTMPyP group was significantly lower than CA group(P<0.05).The levels of SOD,GSH-Px,LAMP2 and Cathepsin D in the CA group were significantly lower than those in the sham group(P<0.05),and the MnTMPyP group was significantly higher than the CA group(P<0.05).Conclusion Oxidative stress inhibition can alleviate the autophagosome clearance injury and lysosomal dysfunction caused by CA-ROSC and inhibit neuronal death.
作者
刘润武
马海芬
徐秦英
LIU Run-wu;MA Hai-fen;XU Qin-ying(Department of Emergency,Qinghai Women and Children Hospital,Xining Qinghai 810007,China;Department of Pathology,Qinghai Institute of Health Sciences,Xining Qinghai 810000,China;Department of Chemistry,Qinghai Institute of Health Sciences,Xining Qinghai 810000,China.)
出处
《临床和实验医学杂志》
2019年第20期2136-2139,共4页
Journal of Clinical and Experimental Medicine
基金
青海省创新平台建设专项项目(编号:2019-0401-ZJC-0014)
