活络效灵丹拆方对乳鼠心肌细胞缺氧/复氧损伤的保护作用研究  

Protective effects of "Huoluo Xiaoling Pellet" on hypoxia/reoxygenation-induced cardiomyocytes injury in neonatal rats

作　　者：韩向东[1] 赵妍[1] 黄云枫[1] 仰薇[1] 

机构地区：[1]上海中医药大学中药学院,上海201203

出　　处：《上海中医药杂志》2012年第3期65-68,共4页Shanghai Journal of Traditional Chinese Medicine

基　　金：上海市教委重点学科建设项目(Z09011);上海市卫生局中医药科研基金项目(2010223)

摘　　要：目的探讨活络效灵丹含药血清对乳鼠心肌细胞缺氧/复氧(H/R)损伤的保护作用及其机制。方法取体外培养的乳鼠心肌细胞,建立缺氧/复氧心肌细胞损伤模型。实验分为正常组、模型组(H/R组)、活络效灵丹拆方组和阳性药对照组(单硝酸异山梨酯组)。用MTT比色法检测活络效灵丹对H/R损伤心肌细胞存活率和细胞上清液中LDH漏出量的影响,优选活络效灵丹的最佳配伍组合,并测定各组细胞浆丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性;用RT-PCR法检测心肌细胞Bcl-2 mRNA、Bax mRNA的表达。结果实验优选出活络效灵丹全方、丹参当归配伍、丹参乳香配伍、丹参当归乳香配伍为最佳配伍。与正常组比较,模型组MDA含量升高、SOD活性降低(P<0.01),细胞Bcl-2 mRNA表达降低、Bax mRNA表达增高(P<0.01);与模型组比较,优选的活络效灵丹各配伍组胞浆SOD活性增高、胞浆MDA含量减少(P<0.01),心肌细胞Bax mRNA表达减少、Bcl-2 mRNA表达增高(P<0.01)。结论活络效灵丹对H/R损伤的心肌细胞具有保护作用,其作用机制可能与清除氧自由基、抗脂质过氧化、上调Bcl-2 mRNA和下调Bax mRNA表达等有关。Objective To study the protective effects of medicated serum with ＂Huoluo Xiaoling Pellet＂ on hypoxia/reoxygenation （H/R） -induced cardiomyocytes injury in neonatal rats. Methods Myoeardial hypoxia/reoxygenation model was established on cultured primary myocardial cells of neonatal rats. The myocardial cells were divided into following groups： normal group, model group （H/R group） , positive control group （Isosorbide M ononitrate） and ＂Hnoluo Xiaoling Pellet＂ （HLXLD） groups. The protective role of HLXLD in eardiocytes survival rate was detected with MTF colorimetric method after H/R injury. The lactate dehydrogenase （LDH） activity in the supernatant, the malondialchehyche （MDA） content and the superoxide dismutase （SOD） activity in the supernatant were measured. The expressions of Bcl-2 mRNA and Bax mRNA were detected by RT-PCR. Results Four herbal compositions of ＂ Huoluo Xiaoling Pellet＂ were screened. Compared with the normal group, MDA eonient was increased and SOD activity was decreased in the model group（P 〈0.01 ） ; the expression of Bcl-2 mRNA was decreased and Bax mRNA was increased in the model group（P 〈0.01 ）. Compared with the model group, in the screened HLXLD groups, the SOD activity increased and MDA content was decreased （P 〈 0.01 ） ;and the expression of Bel-2 mRNA was increased and Bax mRNA was decreased （P 〈 0.01 ）. Conclusion ＂Huoluo Xiaoling Pellet＂ can prevent cardiomyocytes from hypoxia/reoxygenation injury in neonatal rats by eliminating oxygen radical, fighting lipid peroxidation, upregulating Bcl-2 mRNA expression and down-regulating Bax mRNA.

关 键 词：活络效灵丹 心肌细胞 缺氧/复氧损伤 抗氧化 基因表达 

分 类 号：R285.5[医药卫生—中药学]