活络效灵丹对乳鼠缺氧/复氧损伤心肌细胞的保护作用研究

Protective Effects of Huoluo Xiaoling Dan Against Hypoxia/Reoxygenation Injury in Neonatal Rat Cardiomyocytes

目的探讨活络效灵丹含药血清对乳鼠心肌细胞缺氧/复氧(hypoxia/reoxygenation,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H/R心肌细胞损伤模型,实验分为4组:正常血清对照组、模型组(H/R组)、活络效灵丹(HXP)血清组和单硝酸异山梨酯(ISMN)血清组,采用MTT比色法检测活络效灵丹对H/R损伤心肌细胞存活率的影响,测定细胞上清液乳酸脱氢酶(LDH)的漏出量以及胞浆丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性,并采用流式细胞仪检测活络效灵丹对H/R损伤心肌细胞凋亡的影响。结果与正常组比较,模型组细胞存活率明显降低,上清液中LDH漏出量和胞浆MDA含量升高,SOD活性降低,细胞凋亡率显著增加(P<0.01);与模型组比较,活络效灵丹含药血清可明显提高细胞存活率,增加胞浆SOD活性,减少上清液中LDH漏出量和胞浆MDA含量,并降低细胞的凋亡率(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,其作用可能与其抗脂质过氧化作用有关。

Objective To study the protective effects of medicated serum with Huoluo Xiaoling Dan（HXD） on hypoxia/ reoxygenation （H/R） injury in neonatal rat cardiomyocytes. Methods H/R cardiomyocyte model was established on cultured primary myocardial cells of neonatal rats. The myocardial cells were divided into four groups： normal control group, H/R group, serum containing HXD group and serum containing isosorbide mononitrate（ISMN）. The protective effect of HXD on the survival rate of cardiocytes was detected with MTY colorimetric method after H/R injury. The cell apoptosis was determined by flow cytometry. Moreover, the lactate dehydrogenase（LDH） volume leaking out from the supernatant, the malondialchehyche （MDA） content and the superoxide dismutase （SOD） activity in the supernatant were measured. Results Compared with the normal control group, the cardiocytes survival rate and the SOD activity in H/R group decreased, while LDH volume, MDA content and apoptotic rate increased （P 〈 0.01 ）. Compared with H/R group, the cardiocytes survival rate and the SOD activity in HXD group significantly increased, while LDH vol- ume, MDA content and apoptotic rate decreased（P 〈 0.01 ）. Conclusion Huoluo Xiaoling Pills can protect cardiomy- ocytes from H/Rinduced injury, and the mechanism may depend on its effect against oxygen free radical,