昆布多糖硫酸酯对H---_2O_2诱导乳鼠心肌细胞损伤的保护作用

Protective effects of laminaria polysaccharide sulfate on cardiomyocytes of neonatal rats injuried by hydrogen peroxide(H_2O_2)

目的研究昆布多糖硫酸酯(LAPS)对过氧化氢(H2O2)诱导乳鼠心肌细胞氧化损伤的保护作用。方法分次消化法分离原代培养的乳鼠心肌细胞,建立H2O2诱导心肌细胞损伤模型;四甲基偶氮唑盐(MTT)法检测不同浓度LAPS对心肌细胞的保护作用;试剂盒检测LAPS对细胞乳酸脱氢酶(LDH)、肌酸磷酸激酶(CK)、丙二醛(MDA)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)及谷胱甘肽过氧化物酶(GSH-Px)含量的影响;利用荧光探针二氯二氢荧光素-乙酰乙酸酯(DCFH-DA)检测细胞内活性氧(ROS)含量。结果100μmol/L的H2O2能明显造成心肌细胞损伤,LAPS能改善H2O2所造成的心肌细胞损伤;LAPS能够明显降低的过氧化损伤心肌细胞外液中LDH和CK含量、明显降低过氧化损伤心肌细胞内MDA和活性氧(ROS)的含量、明显提高过氧化损伤心肌细胞内SOD、GSH-Px和CAT的活力。结论 LAPS对心肌细胞的过氧化损伤有保护作用,与其降低或阻断脂质过氧化反应,降低细胞内ROS含量,进而阻断因ROS对细胞造成的损害,及时修复受损细胞有着密切关系。

Objective To observe the protective effects of laminaria polysaccharide sulfate（LAPS）on the myocardial cells injured by H2O2.Methods Neonatal cardiomyocytes were isolated from 1-3days old Wistar rats by enzyme digestion method and the cellular injury was induced by H2O2.Cell viability was determined by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyl tetrazolium（MTT）assay.The contents of dehydrogenase（LDH）,creatine kinase（CK）,malonaldehyde（MDA）,surperoxide dismutase（SOD）,glutathione peroxidase（GSH-Px）and catalase（CAT）in the medium or cells were measured.The content of intracellular reactive oxygen species（ROS）was detected by fluorescent probe DCFH-DA.Results 100μmol/L H2O2 could induce the damage of cardiomyocytes.LAPS could mitigate the injury of H2O2 on cardiomyocytes.The releases of LDH and CK were decreased in the supernatant,the contents of MDA and ROS in cells were decreased,the viabilities of SOD,CAT and GSH-Px were all increased after oxidative damaged cells were treated with LAPS.Conclusion LAPS showed protective effects on cardiomyocytes of neonatal rats injuried by H2O2,which closely related to the blocking lipid peroxidation,the declining intercellular ROS effects and the repairing of damaged cells timely.