蕨麻多酚对大鼠心肌细胞缺氧损伤的保护作用研究  

Protective effect of polyphenols from potentilla anserine L.against hypoxia injury in rat cardiomyocytes

作　　者：孙富增 李霞[1] 李雅潇[1] 肖泽云[1] 张冠华[1] 陈立军 李灵芝[1,4] SUN Fu-zeng;LI Xia;LI Ya-xiao;XIAO Ze-yun;ZHANG Guan-hua;CHEN Li-jun;LI Ling-zhi(Department of Military Medicine,Logistic University of PAP,Tianjin 300309,China)

机构地区：[1]武警后勤学院军事药学教研室,天津300309 [2]武警江苏总队医院药剂科,江苏扬州210024 [3]武警后勤学院人体机能学教研室,天津300309 [4]天津市职业与环境危害防制重点实验室,天津300309

出　　处：《武警后勤学院学报（医学版）》2021年第4期1-5,共5页Journal of Logistics University of PAP（Medical Sciences）

基　　金：国家自然科学基金项目(8147182);军队后勤科研重点项目(BWJ19J002);武警后勤学院课题(WHJ202009)。

摘　　要：【目的】研究蕨麻多酚对缺氧损伤心肌细胞的保护作用。【方法】建立大鼠原代培养心肌细胞缺氧损伤模型;实验设对照组、缺氧模型组、蕨麻多酚高、中、低浓度组(0.44、0.22、0.11 mg/ml)及复方丹参片组(3 mg/ml)。采用MTT法检测细胞代谢活力;比色法测定细胞外乳酸脱氢酶(lactate dehydrogenase,LDH)、肌酸激酶(creatine kinase,CK)、一氧化氮(NO)水平以及心肌细胞超氧化物歧化酶(superoxide dismutase,SOD)活性和丙二醛(maondialdehyde,MDA)含量;酶联免疫方法检测心肌细胞内皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)和诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)蛋白水平;DAPI染色方法观察心肌细胞形态学变化。【结果】蕨麻多酚各浓度组显著提高缺氧损伤细胞内SOD活性(P<0.01或P<0.05),减少MDA的产生(P<0.05);下调iNOS蛋白水平,上调eNOS蛋白水平(P<0.01),促进NO合成和分泌(P<0.01或P<0.05);提高缺氧心肌细胞代谢活力,保护细胞结构,减少LDH和CK的外漏(P<0.01或P<0.05)。【结论】蕨麻多酚对心肌细胞缺氧损伤具有保护作用,该作用与其抗自由基氧化能力有关。【Objective】To study the protective effect of polyphenols from potentilla anserine L.on the cardiomyocytes injured by hypoxia.【Methods】Hypoxia injury model of primary cultured myocardial cells in rats was established.A control group,an anoxic model group,groups of polyphenols(0.44 mg/ml,0.22 mg/ml,0.11 mg/ml)and a compound Danshen tablet group(3 mg/ml)were set up in the experiment.The myocardiocytes of rats were exposed to hypoxia in vitro.The metabolic activity of the cells were determined via MTT assay.The activities of lactate dehydrogenase(LDH),creatine kinase(CK)and the level of nitric oxide(NO)in culture medium and the activity of superoxide dismutase(SOD)and the level malondialdehyde(MDA)in cells were measured by spectrophotometry.The levels of endothelial nitric oxide synthase(eNOS)and inducible nitric oxide synthase(iNOS)protein in cardiomyocytes were detected by enzyme-linked immunosorbent assay(ELISA).The morphological changes of cardiomyocytes were observed by DAPI staining.【Results】Under hypoxia,the polyphenols from potentilla anserine L.significantly increased SOD activity in cells(P<0.01 or P<0.05),decreased MDA production(P<0.05),down-regulated iNOS protein level,up-regulated eNOS protein level(P<0.01),promoted NO synthesis and secretion(P<0.01 or P<0.05),increased cardiomyocyte vitality.The cell structure was protected and the leakage of LDH and CK was reduced(P<0.01 or P<0.05).【Conclusion】The polyphenol fraction has a protective effect on myocardial hypoxia injury,which is related to its anti-free radical oxidation ability.

关 键 词：蕨麻多酚 缺氧 抗氧化 心肌细胞 一氧化氮 一氧化氮合酶 

分 类 号：R285.5[医药卫生—中药学]