摘要
目的探讨胰高血糖素样肽-1(GLP-1)对乳鼠心肌细胞缺氧/复氧(H/R)损伤中的抗氧化作用。方法取出生1~3d的SD大鼠乳鼠心脏,采用差速贴壁法分离并培养其心肌细胞,通过对乳鼠心肌细胞H/R损伤模拟缺血再灌注。实验共分为5组,A组:正常对照组;B组:H/R组;C组:缺氧预处理组,先缺氧20 min后复氧20 min,然后进行H/R;D组:GLP-1激动剂组,Exendin-4预处理之后进行H/R;E组:GLP-1拮抗剂组,Exendin-4+Exendin-(9-39)预处理之后进行H/R。各组于复氧后,使用MTT比色法检测各组细胞存活率;乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)、丙二醛(MDA)试剂盒检测各组细胞损伤程度。结果与A组相比较,B组心肌细胞存活率明显降低,培养液中LDH漏出量显著增加,细胞上清液中SOD活性明显降低且MDA含量明显增加,差异具有统计学意义(P<0.01)。与B组相比较,C组和D组存活率明显增加,培养液中LDH漏出量显著减少,细胞中SOD活性明显升高且MDA含量明显降低,差异具有统计学意义(P<0.05),而二组之间差异无统计学意义(P>0.05)。但给予GLP-1拮抗剂后显著降低其保护作用。结论 Exendin-4通过GLP-1R依赖机制减少缺氧造成的氧化应激损伤,进而保护心肌细胞,表明GLP-1具有抗氧化保护作用。
Objective To investigate antioxidant protective effect of glucagon like peptide -1（GLP-1） on the hypoxia / reoxygenation （H/R） injury in neonatal rat cardiomyocytes. Methods The rat heart was isolated from 1 to 3 days old SD rats, and the cardiomyocytes were isolated and cultured by differential adherence method. The myocardial ischemiareperfusion injury was simulated by H/R damage. The experiments were divided into 5 groups： group A： normal control group; group B： H/R group; group C： hypoxia preconditioning group, first hypoxia for 20min, and reoxygenation for 20rain, then H/R; group D： GLP-1 agonist group, after Exendin-4 pretreatment, then H/R; group E： GLP-1 antagonist group, after Exendin-4 ＋ Exendin- （9-39） pretreatment, then H/R. After reoxygenation, the cell survival rate of each group was detected by MTT colorimetric assay, and the cell damage degree of each group was detected by LDH, superoxide dismutase （SOD） and malondialdehyde （MDA）. Results Compared with A group, myocardial cell survival rate in B group was significantly decreased, LDH in culture fluid leakage in the supernatanl was significantly increased, SOD activity decreased and MDA content increased significantly, the difference was statistically significant （P 〈 0.01）. Compared with B group, the survival rate of C group and D group was significantly increased, the LDH leakage in the culture medium significantly reduced, the activity of SOD in cells was significantly inereased and the content of MDA was significantly reduced, the difference was statistically significant （P 〈 0.05）, but there was no significant difference be tween the two groups （P 〉 0.05）. However, the protective effect was significantly decreased after giving GLP-1 antagonist. Conclusion Exendin-4 can reduce the oxidative stress injury induced by hypoxia and protect the myocardial cells through GLP-IR dependent mechanism, which indicates that GLP-1 has the protective effect of antioxidation.
出处
《中国医药导报》
CAS
2017年第4期12-15,共4页
China Medical Herald
基金
国家自然科学基金(81570210)
