四逆汤水提物对缺氧-复氧损伤心肌细胞的保护作用

Protective Effect of Aqueous Extract of SiniTang on Injuried Cardiac Muscle Cell Induced by Hypoxia- Reoxygenation

目的:观察四逆汤水提物(Sinidecoction,SND)对培养乳鼠心肌细胞在缺氧复氧条件下的直接保护作用。方法:动态采集并分析正常对照组、缺氧复氧组、四逆汤水提物8.0、17.9、40.0、89.4、200μg/ml给药组心肌细胞在缺氧前,缺氧30、60、90、120、150、180min以及复氧30min和60min细胞搏动频率及收缩面积的变化;比色法测定培养液中LDH;使用MTT法测定心肌细胞存活量。结果:心肌细胞搏动频率及收缩幅度随着缺氧时间延长而逐渐降低,四逆汤水提物给药组(除8.0μg/ml组外)较缺氧复氧组搏动维持时间长,且200.0μg/ml组在复氧后能恢复搏动。缺氧3h缺氧复氧组乳酸脱氢酶(LDH)漏出率较正常对照组升高(P<0.05);除8.0μg/ml组外,四逆汤水提物不同浓度组LDH漏出率较缺氧复氧组降低,存在浓度-效应关系。复氧1h,各给药组LDH漏出率也较缺氧复氧组低。缺氧复氧组与正常对照组比较,存活的心肌细胞减少(P<0.01);给药组心肌细胞存活量较高,呈浓度-效应关系。结论:四逆汤水提物可延长缺氧状态下心肌细胞的搏动时间,减缓收缩力的衰减,减少细胞膜损伤及提高缺氧复氧心肌细胞的存活量,表现出对心肌细胞的直接保护作用。

Objective: To investigate the direct protective effect of SiniTang (SNT) on cultured myocardial cells of neonatal rats injuried by hypoxia and reoxygenation. Methods: The contracting frequency and content of primary-cultured myocardial cells sampled from different groups (control, oxygen and glucose deprivation model, and theated groups with 8. 0, 17. 9, 40, 89. 4, 200μg/mL SNT) were determined by TV recording system and computer image analysis system at different time (hypoxia for 30min, 60min, 90min, 120min, 150min, 180min and reoxygenation for 30min, 60min). The LDH activity in culture media was measured colorimetrically. The quantity of surviving myocardial cells was measured with MTT reduction method. Results: The contraction frequency and range of cultured myocardial cells decreased during hypoxia, the myocardial cells'pulsation of the groups treated with SNT was kept longer than the oxygen and glucose deprivation model, and the myocardial cells treated with SNT of 200μg/mL pulsed again after reoxygenation. After hypoxia for 3 hours, the LDH release of oxygen and glucose deprivation model was significant higher than that of the control ( P < 0. 05 ) ; the LDH release of the groups treated with SNT ( except the group 8. 0μg/mL) was lower than that of oxygen and glucose deprivation model, and there was a positive correlation between SNT concentration and the decrease of the LDH release. After reoxygenation, the LDH release of the groups treated with SNT was lower than that of oxygen and glucose deprivation model. The quantity of surviving myocardial cellsof oxygen and glucose deprivation model was significantly lower than that of the control (P <0. 01) ; the quantity of surviving myocardial cells of the groups treated with SNT was higher than that of oxygen and glucose deprivation model, and there was a positive correlation between SNT concentration and the increase of the quantity of surviving myocardial cells. Conclusion: SiniTang could keep myocardial cells pulsing for a longer time than oxygen and glucose deprivation model and reduced the decrease of contracting power during hypoxia, protect cell membrane and increase the survival of myocardial cells injured by hypoxia/reoxygenation. SiniTang might have a direct protective effect on cultured myocardial cells injuried by hypoxia and reoxygenation.