白藜芦醇通过调控自噬减轻乳大鼠心肌细胞低氧复氧损伤  

Resveratrol mitigates hypoxia/reoxygenation injury in myocardial cells of newborn rats via autophagy regulation

作　　者：张金芳 刘梦思 孙秀玉 刘立亚[2] 李志勇 黄秀兰 ZHANG Jin-fang;LIU Meng-si;SUN Xiu-yu;LIU Li-ya;LI Zhi-yong;HUANG Xiu-lan(School of Pharmacy,Minzu University of China,Beijing 100081,China;College of Life and Environmental Sciences,Minzu University of China,Beijing 100081,China)

机构地区：[1]中央民族大学药学院,北京100081 [2]中央民族大学生命与环境科学学院,北京100081

出　　处：《中国药理学与毒理学杂志》2020年第8期584-590,共7页Chinese Journal of Pharmacology and Toxicology

基　　金：国家自然科学基金(81173656);国家自然科学基金(81470182)。

摘　　要：目的探讨白藜芦醇(Res)对低氧复氧(H/R)损伤心肌细胞的保护作用。方法采用三气培养箱法制备乳大鼠心肌细胞H/R损伤模型(H12 h/R12 h)。乳大鼠心肌细胞分为5组:细胞对照组、H/R模型组、模型+Res 5和20μmol·L^-1组(Res预处理细胞12 h,然后H12 h/R12 h)和模型+Res 20μmol·L^-1+氯喹(CQ)10μmol·L^-1组(Res预处理细胞12 h,然后含CQ的培养基H12 h/R12 h);CCK-8法检测心肌细胞存活率,比色法测定心肌细胞乳酸脱氢酶(LDH)漏出率,AnnexinⅤ/PI双染法检测细胞凋亡,免疫荧光法检测心肌细胞内微管相关蛋白1轻链3(LC3)阳性细胞率,Western印迹法检测心肌细胞Beclin 1,LC3Ⅱ和P62蛋白表达水平。结果与细胞对照组比较,H/R组心肌细胞存活率下降(P<0.01),细胞凋亡率和LDH漏出率上升(P<0.01),LC3阳性细胞率和Beclin 1表达水平无明显变化,LC3Ⅱ和P62表达水平下降(P<0.05)。与H/R组比较,Res 5和20μmol·L^-1可升高心肌细胞存活率(P<0.05),降低凋亡率和LDH漏出率(P<0.01),LC3阳性细胞率增高(P<0.05),Beclin 1,LC3Ⅱ和P62蛋白表达水平增加(P<0.05)。与模型+Res 20μmol·L^-1组比较,加用CQ组心肌细胞存活率下降(P<0.05),凋亡率升高(P<0.05),LDH漏出率增高(P<0.01),Beclin 1,LC3Ⅱ和P62蛋白水平和LC3阳性细胞率无明显变化。结论Res对H/R损伤心肌细胞具有保护作用,其机制可能与活化自噬有关。OBJECTIVE To explore the protective effects of resveratrol(Res)against hypoxia/reox⁃ygenation injury(H/R)of myocardial cells.METHODS A newborn rat cardiomyocyte H/R model was established using the tri-gas incubator method(H for 12 h,R for 12 h).Myocardial cells of newborn rats were divided into 5 groups:cell control group,H/R model group,model+Res 5 and 20μmol·L^-1 group(cardiomyocytes were pretreated with Res for 12 h,and then processed with H12 h/R12 h)and model+Res 20μmol·L^-1+chloroquine(CQ)10μmol·L^-1 group(myocardial cells were pretreated with Res for 12 h,and then processed with H12 h/R12 h with medium containing CQ).The cell viability was measured by CCK-8 assay.The leakage rate of lactate dehydrogenase(LDH)was determined by LDH kit.The apop⁃tosis was detected by AnnexinⅤ/PI double staining.The rate of intracellular microtubule associated protein 1 light chain 3(LC3)positive cells was observed by immunofluorescence.The expressions of autophagy-related proteins such as Beclin 1,LC3Ⅱand P62 were evaluated by Western blotting.RESULTS Compared with cell control group,the cell viability in H/R group was significantly decreased(P<0.01),the apoptosis rate and leakage rate of LDH were obviously increased(P<0.01),the rate of LC3 positive cells and the protein expression of Beclin 1 did not change significantly,but LC3Ⅱand P62 protein expressions were decreased(P<0.05).Compared with H/R group,the cell viability of Res 5 and 20μmol·L^-1 group was increased(P<0.05),the apoptosis rate and leakage rate of LDH were all de⁃creased(P<0.01),the rate of LC3 positive cells was increased(P<0.05),and the protein expressions of Beclin 1,LC3Ⅱand P62 were increased(P<0.05).Compared with Res 20μmol·L^-1 group,the cell viability of Res 20μmol·L^-1+CQ 10μmol·L^-1 group was decreased(P<0.05),the apoptosis rate and leakage rate of LDH were increased(P<0.05),but there was no significant difference in protein expres⁃sions of Beclin 1,LC3Ⅱand P62 or in the rate of LC3 positive cells.CONCLUSION Res could pr

关 键 词：白藜芦醇 心肌细胞 低氧复氧损伤 自噬 

分 类 号：R285.5[医药卫生—中药学]