龙牙楤木总皂苷对H_2O_2诱导乳鼠心肌细胞损伤的保护作用  

Protective effects of Aralosides(AS) against H_2O_2-induced injury in neonatal rat cardiomyocytes

作　　者：孙桂波[1] 王敏[1] 高蒙蒙[1] 徐惠波[2] 孙晓波[1] 

机构地区：[1]中国医学科学院药用植物研究所,北京100193 [2]吉林省中医药科学院,吉林长春130021

出　　处：《中国药理学通报》2013年第6期773-777,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81173589);国家重大新药创制专项(No 2009ZX09102-104);吉林省自然科学基金项目(No 201015110)

摘　　要：目的研究龙牙楤木总皂苷(AS)对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;MTT法检测不同浓度AS对细胞保护作用的量效、时效关系;倒置显微镜下观察AS对细胞形态学和细胞搏动频率的影响;试剂盒检测乳酸脱氢酶(LDH)的漏出量及丙二醛(MDA)含量,检测过氧化氢酶(CAT)、超氧化物歧化酶(SOD)及谷胱甘肽过氧化物酶(GSH-Px)活力。结果不同浓度H2O2处理心肌细胞2 h,100～400μmol.L-1对心肌细胞具有明显的损伤作用,呈剂量依赖性;LDH漏出量增加,MDA含量升高,SOD、CAT、GSH-Px活力明显降低。而12.5～25 mg.L-1AS预处理8、12 h可明显地减轻损伤,有效保护心肌细胞。明显减少LDH的漏出,降低MDA含量,提高SOD、CAT、GSH-Px活力;12.5～25 mg.L-1 AS于给药后4h明显减慢心肌细胞的搏动频率,并呈剂量依赖性,表明在该剂量范围内AS具有明显的负性频率效应。结论 AS对H2O2致心肌细胞氧化损伤具有明显的保护作用,呈时间、剂量依赖性。其作用机制可能与其清除自由基,提高心肌细胞的抗氧酶活力有关;同时提示AS的心肌保护作用可能与其减少心肌做功、降低心肌耗氧量、增加心肌细胞稳定性有关。Aim To investigate the protective effects of Aralosides against H2O2-induced injury in neonatal rat eardiomyocytes. Methods A model of H2O2-indueed injury in neonatal rat cardiomyocytes was established. Cell viability was determined by 3-（4,5-dimethylthia- zol-2-yl） -2,5-diphenyl tetrazolium （MTT） assay. The changes of cell morphology and the myocardial beating rate were detected with the invert microscope. The su- pernates were collected to measure the LDH levels and the cells were harvested to determine the MDA levels and the activities of SOD, CAT and GSH-PX with the respective detection kits according to the manufactur- er＇s instructions. Results 100-400 μmol.L-1 of H2O2 could cause a significant injury in the neonatal rat eardiomyocytes in a dose-dependent manner. How- ever, Aralosides treatment could attenuate H2O2-in-duced decrease in cell viability of myocardial cells, ac- tivities of SOD, CAT and GSH-PX, as well as increase in the levels of LDH and MDA. The beating rate of myocardial cells got slower in a dose-dependent man- ner, indicating that the myocardial cells were apparent- ly effected by negative frequency. Conclusion Aralo- sides exerts protective effects against H2O2-induced in- jury in myocardial cells in dose- and time-dependent manners. The mechanism of these protective effects may be due to its scavenging free radicals and increas- ing the activities of antioxidant enzymes, as well as de- creasing myocardial performance and oxygen consump- tion and increasing the stabilization of myocardial cells.

关 键 词：龙牙楤木总皂苷 H2O2 乳鼠心肌细胞 抗氧化 搏动频率 自由基 

分 类 号：R332[医药卫生—人体生理学] R284.1[医药卫生—基础医学]