富马酸氯马斯汀对大鼠心肌细胞缺氧复氧时TLR4/PI3K/Akt信号通路的影响  

Effect of clemastine fumarate on TLR4/PI3K/Akt signaling pathway during hypoxia-reoxygenation in rat cardiomyocytes

作　　者：延茹 岳峰 刘永新 袁晓晓 孙美艳 张蕊[1] 隽兆东[1] 黄亚茹 沈吉喆 Yan Ru;Yue Feng;Liu Yongxin;Yuan Xiaoxiao;Sun Meiyan;Zhang Rui;Juan Zhaodong;Huang Yaru;Shen Jizhe(Department of Anesthesiology,Weifang Medical University Shandong Provincial Medicine and Health Key Laboratory of Clinical Anesthesia,Weifang 261053,China)

机构地区：[1]潍坊医学院麻醉学系,山东省医药卫生临床麻醉重点实验室,261053

出　　处：《中华麻醉学杂志》2019年第5期610-612,共3页Chinese Journal of Anesthesiology

基　　金：山东省自然科学基金(ZR2017LH002,ZR2017MH066)

摘　　要：目的评价富马酸氯马斯汀对大鼠心肌细胞缺氧复氧时Toll样受体4/磷脂酰肌醇-3-激酶/丝氨酸-苏氨酸激酶(TLR4/PI3K/Akt)信号通路的影响。方法体外培养大鼠H9C2心肌细胞,以1×10^5个/ml接种于培养孔或培养瓶,采用随机数字表法分为3组(n=11):对照组(C组)、缺氧复氧组(H/R组)和富马酸氯马斯汀组(CF组)。在低糖DMEM培养基-5%CO2-95%N2中缺氧4 h,复氧4 h制备心肌细胞缺氧复氧损伤模型。于复氧4 h时采用CCK-8法检测细胞活力;透射电镜观察超微结构;Western blot法检测TLR4、PI3K、磷酸化Akt(p-Akt)和caspase-3的表达,免疫荧光法检测TLR4、PI3K和caspase-3的表达。结果与C组比较,H/R组细胞活力降低,TLR4和caspase-3表达上调,PI3K和p-Akt表达下调(P<0.05);与H/R组比较,CF组细胞活力升高,TLR4和caspase-3表达下调,PI3K和p-Akt表达上调(P<0.05),线粒体损伤减轻。结论富马酸氯马斯汀减轻大鼠心肌细胞缺氧复氧损伤的机制可能与抑制TLR4表达,激活PI3K/Akt信号通路有关。Objective To evaluate the effect of clemastine fumarate on Toll-like receptor 4/phosphatidylinositol-3-kinase/serine-threonine kinase(TLR4/PI3K/Akt)signaling pathway during hypoxia-reoxygenation(H/R)in rat cardiomyocytes.Methods H9C2 cells of rats cultured in vitro were seeded in culture wells or dishes at a density of 1×10^5 cells/ml and divided into 3 groups(n=11 each)by using a random number table method:control group(group C),H/R group and clemastine fumarate group(CF group).Cardiomyocytes were exposed to 5%CO2-95%N2 in a low-glucose DMEM medium at 37℃for 4 h followed by 4 h reoxygenation.At 4 h of reoxygenation,the cell viability was detected by CCK-8 assay,the ultrastructure was observed with a transmission electron microscope,the expression of TLR4,PI3K,phosphorylated Akt(p-Akt)and caspase-3 was detected by Western blot,and the expression of TLR4,PI3K and caspase-3 was detected by immunofluorescence.Results Compared with group C,the cell viability was significantly decreased,the expression of TLR4 and caspase-3 was up-regulated,and the expression of PI3K and p-Akt was down-regulated in group H/R(P<0.05).Compared with group H/R,the cell viability was significantly increased,the expression of TLR4 and caspase-3 was down-regulated,the expression of PI3K and p-Akt was up-regulated(P<0.05),and the mitochondrial damage was significantly attenuated in group CF.Conclusion The mechanism by which clemastine fumarate alleviates H/R injury to rat cardiomyocytes may be related to inhibiting TLR4 expression and activating PI3K/Akt signaling pathway.

关 键 词：组胺H1拮抗剂 心肌再灌注损伤 Toll样受体4 磷酸肌醇3-激酶类 蛋白质丝氨酸苏氨酸激酶 

分 类 号：R73[医药卫生—肿瘤]