Protective Effects Of Huoluoxiaolingdan And Its Series Against Hypoxia/Reoxygenation Injury In Neonatal Rat Cardiomyocytes

Huoluoxiaolingdan is a classic decoction, which has powerful curative effects on blood circulation, dredging collaterals, relieving pain. In recent years, Huoluoxiaolingdan has already have a clinical use on coronary heart disease. But to date the study of myocardial ischemia and reperfusion is still less.To study the protective effects of medicated serum with Huoluoxiaolingdan and its series on hypoxia/reoxygenation (H/R) injury in neonatal rat cardiomyocytes, myocardial H/R model was established on cultured primary myocardial cells of neonatal rat. The isosorbide mononitrate has been used as positive control. The key results are shown as below:1. Evaluating the protective effects of Huoluoxiaolingdan against H/R injury in neonatal rat cardiomyocytes. The protective role of Huoluoxiaolingdan in cardiocytes survival rate was detected with MTT colorimetric method after H/R injury. The cell apoptosis was determined by flow cytometry. Moreover, the lactate dehydrogenase (LDH) activity in the supernatant, the malondialchehyche (MDA) content and the superoxide dismutase (SOD) activity in the supernatant were measured. Then the cell apoptosis was determined by flow cytometry. The results showed that Huoluoxiaolingdan has a significant protective effects on H/R injury cardiomyocytes.2. Preliminaryly screening the series of Huoluoxiaolingdan by the protective effects on H/R injury cardiomyocytes. The protective role of Huoluoxiaolingdan in cardiocytes survival rate was detected with MTT colorimetric method after H/R injury. Meanwhile, the lactate dehydrogenase (LDH) activity in the supernatant was measured. The results showed that the medicated serum of Huoluoxiaolingdan, danshendanggui, danshenruxiang and danshendangguiruxiang have significant protective effects.3. Investigating the action mechanism of protective effects on H/R injury cardiomyocyte. The malondialchehyche (MDA) content and the superoxide dismutase (SOD) activity in the supernatant were measured by colorimetric method. And RT-PCR was used to test the expression of gene-related apoptosis Bcl-2 mRNA and Bax mRNA. The results showed that the reducing of oxygen free radical and inhibiting myocardial apoptosis may be the possible mechanisms.4. Angelica Sinensis and Salvia Miltiorrhiza in Huoluoxiaolingdan play an leading role on the protection of myocardial cells in antioxidant injury and inhibiting myocardial apoptosis gene expression.In summary, Huoluoxiaolingdan and its series can protect cardiomyocytes from H/R-induced injure and exhibit a potentional application value on the treatment ofmyocardial ischemia and reperfusion.