δ阿片受体激活对过氧化氢损伤的心肌细胞的保护作用  

Protection of δ-opioid receptor stimulation against injured myocardial cells by hydrogen peroxide

作　　者：李炯[1] 王忠彦[2] 于敏[3] 

机构地区：[1]滨州医学院卫生管理学院 [2]滨州医学院新闻中心 [3]滨州医学院药学院,山东烟台264003

出　　处：《中国药理学与毒理学杂志》2009年第6期431-435,共5页Chinese Journal of Pharmacology and Toxicology

摘　　要：目的研究δ阿片受体激活剂D-丙(2)-D-亮-(5)-脑啡肽(DADLE)对过氧化氢(H2O2)损伤的心肌细胞的保护作用及其机制。方法分离乳大鼠心肌细胞,培养48h后分为正常对照、H2O2(200μmol.L-1)、H2O2+DADLE(1μmol.L-1)、H2O2+DADLE+纳曲吲哚(10μmol.L-1)和H2O2+DADLE+U0126(10nmol.L-1)组,继续培养48h。用[3H]TdR掺入法检测心肌细胞增殖反应,流式细胞仪检测心肌细胞凋亡百分率,乳酸脱氢酶(LDH)活性测定试剂盒测定培养上清LDH活性,硫代巴比妥酸显色法测定细胞内丙二醛(MDA)含量,黄嘌呤氧化酶法测定细胞内超氧化物歧化酶(SOD)活性,Western蛋白印迹法检测细胞外信号调节激酶磷酸化(p-ERK)水平。结果①与正常对照组比较,H2O2组心肌细胞[3H]TdR掺入值明显降低,细胞凋亡率升高;培养上清LDH活性和MDA含量明显增加,SOD活性和Ap-ERK/AERK的比值降低。②与H2O2组比较,DADLE可使心肌细胞[3H]TdR掺入值升高,细胞凋亡率下降;培养上清LDH活性和MDA含量降低,SOD活性和Ap-ERK/AERK的比值升高。③分别加入δ阿片受体拮抗剂纳曲吲哚和ERK拮抗剂U0126,DADLE对上述指标的逆转作用被抑制。结论δ阿片受体激活对H2O2损伤的心肌细胞具有保护作用,其机制可能与其增强心肌细胞的抗氧化功能及促进ERK磷酸化有关。AIM To study protective effect of [D-Ala2,D-Leu5]-enkephalin（DADLE） against hydrogen peroxide（H2O2） induced myocardial cell injury and its possible mechanisms.METHODS Myocardial cells were isolated from neonatal rats and cultured for 48 h.Then the cells were randomly assigned into normal control,H2O2（200 μmol·L-1）,H2O2＋DADLE（1 μmol·L-1）,H2O2＋DADLE ＋naltrindole（10 μmol·L-1） and H2O2＋DADLE ＋U0126（10 nmol·L-1） groups and cultured for another 48 h.[3H]TdR incorporation assay and flow cytometry were used to measure the cell proliferation and apoptosis rate.The lactate dehydrogenase（LDH） activities in culture supernatant measured by using LDH activity kit.The superoxide dismutase（SOD） activity and malondialdehyde（MDA） content in cells were measured with xanthine oxidase method and color reaction of thiobarbituric acid,respectively.The expressions of extracellular signal-regulated kinase（ERK） and phosphorylated-ERK（p-ERK） were observed with Western blot.RESULTS ① Compared with normal control group,the incorporation of [3H]TdR in myocardial cells of H2O2 group was significantly lower,apoptosis rate was higher,LDH activity and MDA content in cells were higher,while SOD activity in cells was lower.In addition,the ratio of IAp-ERK/IAERK was decreased.② Compared with H2O2 group,the incorporation of [3H]TdR in H2O2＋DADLE group was significantly higher,apoptosis rate was lower,LDH activity and MDA content in cells decreased,while SOD activity increased significantly.The ratio of IAp-ERK/IAERK was increased.③ δ-Opioid receptor antagonist naltrindole and ERK antagonist U0126 inhibited this effect of DADLE on the above index changes induced by H2O2.CONCLUSION The δ-opioid receptor has protective effect against H2O2-induced myocardial cell injury,and its possible mechanism may be related to its promotion of antioxide capacity and ERK phosphorylation.

关 键 词：受体 阿片样 δ 肌细胞 心脏 过氧化氢 细胞外信号调节MAP激酶类 超氧化物歧化酶 

分 类 号：R542.22[医药卫生—心血管疾病]