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目的:建立一种同时测定和营安心方中人参皂苷Rg1与丹参酮ⅡA含量的HPLC方法,并对大鼠心肌细胞抗氧化机制进行探讨。方法:采用传统煎煮法提取,HPLC法测定含量[Krosmosil C18柱(4.6mm×200mm,5μm);流动相为乙腈-0.3%磷酸水;检测波长203nm、270nm];采用H2O2建立大鼠胚胎心肌细胞氧化应激损伤模型,分为5组:正常组,模型组,和营安心方高、中、低剂量组,对乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)、一氧化氮(NO)与丙二醛(MDA)进行含量测定。结果:和营安心方中人参皂苷Rg1与丹参酮ⅡA在0.1125-1.8000μg,0.2250-4.0800μg内呈良好的线性关系,人参皂苷Rg1与丹参酮ⅡA的平均含量为6.58mg/g,14.19mg/g。体外研究表明,和营安心方可使H2O2损伤的心肌细胞MTT代谢率提高,LDH含量减少,NO、MDA含量显著下降,SOD含量显著提高(P<0.05,P<0.01)。结论:和营安心方对H2O2损伤心肌具有明显的保护作用,其机制可能与和营安心方中人参皂苷Rg1与丹参酮ⅡA抑制心肌细胞脂质过氧化作用有关。
OBJECTIVE: To establish a HPLC method for the simultaneous determination of ginsenoside Rg1 and tanshinone Ⅱ A in Yingananxinfang, and to explore the mechanism of antioxidation in rat cardiomyocytes. Methods: The contents of [Krosmosil C18 column (4.6 mm × 200 mm, 5 μm), acetonitrile-0.3% phosphoric acid water, detection wavelength of 203 nm and 270 nm] were determined by traditional decoction method and determined by HPLC. Cell oxidative stress injury model was divided into five groups: normal group, model group, and Yinganxinfang high, medium and low dose groups, the effect of lactate dehydrogenase (LDH), superoxide dismutase (SOD) Nitrogen (NO) and malondialdehyde (MDA) were determined. Results: There was a good linear relationship between ginsenoside Rg1 and tanshinone ⅡA in the range of 0.1125-1.8000μg and 0.2250-4.0800μg. The average contents of ginsenoside Rg1 and tanshinone ⅡA were 6.58mg / g and 14.19mg / g. In vitro studies show that Hailinganxin Fang can increase the MTT metabolic rate of cardiomyocytes injured by H2O2, reduce the content of LDH, decrease the content of NO and MDA, and increase the content of SOD significantly (P <0.05, P <0.01). CONCLUSION: Weideanxinxin decoction has significant protective effect on H2O2 injured myocardium, and its mechanism may be related to the inhibitory effects of ginsenoside Rg1 and tanshinone ⅡA on myocardial cell lipid peroxidation.