四逆汤有效部位抗心肌缺血/再灌注损伤神经酰胺机制的研究

Ceramide mechanism of anti-myocardial ischemia/reperfusion injury of effective position in Sini Decoction

目的:探讨四逆汤有效部位对心肌缺血再灌注过程中心肌细胞凋亡及神经酰胺含量的影响。方法:昆明种小鼠,随机分为对照组,模型组,四逆汤有效部位组,用垂体后叶素造模,取小鼠心肌测定SOD活性及MDA含量,通过DNALadder和Dap i荧光染色考察心肌细胞凋亡情况,通过高效薄层层析及薄层层析扫描,用随引标准曲线计算心肌组织中神经酰胺的含量。结果:模型组心肌细胞有凋亡特有的DNA ladder,凋亡指数和神经酰胺含量及MDA含量均比正常组显著升高(P<0.01),SOD活性比正常组显著降低(P<0.01);与模型组相比,四逆汤有效部位组的凋亡指数和神经酰胺含量均显著降低(P<0.01),SOD活性比模型组显著升高(P<0.01);模型组神经酰胺含量与凋亡指数和MDA含量均呈明显正相关(r=0.970,P<0.01;r=0.974,P<0.001)。结论:四逆汤有效部位可在一定程度上降低心肌缺血再灌注时增高的心肌神经酰胺的含量,减少心肌细胞凋亡,从而达到保护心肌的作用。

AIM： To study the effects of Sini Decoction＇s effective component（SNDE） on myocardial apoptosis and ceramide content during myocardial ischemla/reperfusion. METHODS： The mice of Kunming species were randomly divided into 3 groups ： control, myocardial ischemia/reperfusion and SNDE. After inducing myocardial ischemia/reperfusion injury of mice in vivo with pituitrin （Pit）, myocardial SOD activity and MDA content were measured. DNA agarose gel electrophoresis and fluorescent stain of DAPI were done to check up apoptosis. The content of myocardial ceramide （μg/kg） was measured through HPTLC and scan of thin plate. RESULTS： The myocardium of model group has the phenomenon of DNA ladder. The apoptosis index and the ceramide content in model group were higher than those of control group （ P〈0.01 ） . SOD activity of model group was lower than that of control group （P〈0.01 ）. The apoptosis index and the ceramide content of the SNDE group were lower than those of ischemic group. （ P〈0.01 ）. SOD activity of SNDE group was higher than that of model group （ P〈0.01 ）. The apoptosis index and ceramide content in model group were positively correlative （ r=0. 970, P〈0. 001）. The myocardial content ofceramide and MDA were positively correlative too （r=0. 974, P〈0. 001 ）. CONCLUSION： Sini Decoction’s effective component（SNDE） can protect myocardium from decreasing the content of ceramide and reducing myocardium apoptosis.