香青兰提取物对体外培养乳鼠心肌细胞缺氧/复氧损伤的保护作用

Protective effects of dracocephalum moldavica extracts on hypoxia/reoxygenation injury of cardiomyocytes in cultured neonatal rat

目的:研究香青兰提取物对心肌细胞缺氧/复氧(H/R)损伤的保护作用及机制。方法:提取香青兰的有效成分,用原代培养的SD大鼠乳鼠心肌细胞建立H/R的损伤模型,实验分6组:正常对照组、H/R组、总提取物组、氯仿组、乙酸乙酯组及正丁醇组。采用MTT法测定细胞的存活率,乳酸脱氢酶试剂盒(LDH)测定培养液中LDH的含量,黄嘌呤氧化酶法测定细胞内超氧化物歧化酶(SOD)的活性以及硫代巴比妥酸显色法测定活细胞丙二醛(MDA)的含量。结果:各组细胞活力、吸光度值以及LDH、MDA和SOD含量间比较,差异均有统计学意义(F=102.312、51.007、24.136、70.281和14.335,P<0.05)。与正常对照组比较,模型组LDH、MDA的含量及细胞活力明显升高,SOD活性降低(P<0.05);与模型组比较,香青兰提取物各组的细胞活力降低,LDH和MDA含量增加;氯仿和乙酸乙酯组的吸光度值升高;氯仿、乙酸乙酯和正丁醇组的SOD含量增加(P均<0.05)。结论:香青兰提取物对H/R心肌细胞的损伤具有保护作用,此作用可能与抑制脂质过氧化和减少心肌细胞的凋亡有关。

Aim：To investigate the protective effect and the mechanism of dracocephalum moldavica extracts on the damage induced by hypoxia/reoxygenation（H/R） in cultured neonatal rat cardiomyocytes.Methods：The models of hypoxia/reoxygenation were made with the first generation of cultured cardiomyocytes. They were allocated into six groups：normal control group,H/R group （anoxia for 2 hours,reoxygenation for 30 min）,coreopsis total extract group,coreopsis chloroform extract group,coreopsis ethylacetate extract group,and coreopsis n-butanol extract group.The viabilities of cardiomyocytes were envaluted by MTT.The activity of LDH was envaluted by Beckman CX5 promachine.The content of MDA and activity of SOD in cells were envaluted by MDA and SOD kit.Results：The viabilities of cardiomyocytes,absorbance value,LDH,MDA,and SOD were obvious different among the 6 groups（F=102.312,51.007,24.136,70.281,14.335,P0.05）.Compared with normal control groups,the content of LDH,MDA and the cell viability in modol group were increased apparenthy（P0.05） and MDA decreased apparently（P0.05）.Compared with model group,the cell viability decreased apparently in dracocephalum moldavica exeracts groups,the LDH and MDA indeased apparently,the absorbance value increased in coreopsis chloroform extrct group and coreopsis ethylacetate extract group.The SOD increased apparenthy in coreopsis chloroform extracte group,coreopsis ethylacetate extract group and coreopsis n-butanol extract group.Conclusion：Dracocephalum moldavica extracts have the protective effect on hypoxia/reoxygeneration injure of cardiac myocyte in cultured neonatal rat.The mechanism may be in relation to the restraint of surcharge of calcium and reduction of apoptosis.