活血、破血药对动脉粥样硬化大鼠主动脉组织内源性凋亡途径的影响

Effect of Blood-activating Drug and Stasis-breaking Drug on Endogenous Apoptotic Pathway of Aortic Tissue in Atherosclerosis Rats

目的探讨活血药(当归、川芎)、破血药(三棱、莪术)改善动脉粥样硬化的可能作用机制。方法将60只大鼠随机分为空白组、模型组、他汀组、活血组、破血组,空白组8只,其余各组均13只。采用高脂饲料喂饲结合腹腔注射维生素D3法复制动脉粥样硬化大鼠模型。造模成功后,空白组、模型组灌服10 ml/(kg·d)白开水,活血组灌服当归、川芎煎液,破血组灌服三棱、莪术煎液,他汀组灌服阿托伐他汀钙,给药量均为1.8 g/(kg·d),每天1次,连续4周。取主动脉组织,光镜下观察主动脉组织病理变化,免疫组化法检测主动脉组织细胞色素C(Cyt-C)及细胞凋亡诱导因子(AIF)蛋白的表达,原位杂交法检测主动脉组织半胱氨酸天冬氨酸蛋白酶3(Caspase-3)mRNA表达情况。结果与模型组比较,他汀组、活血组及破血组主动脉组织粥样硬化病变均有不同程度减轻,其中他汀组与破血组病变改善最为明显。与空白组比较,模型组主动脉组织Cyt-C、AIF蛋白及Caspase-3 mRNA表达显著增多(P<0.05,P<0.01);与模型组比较,他汀组、破血组主动脉组织Cyt-C、AIF蛋白及Caspase-3 mRNA表达均减少,活血组Cyt-C蛋白、Caspase-3 mRNA表达均减少(P<0.05或P<0.01);破血组主动脉组织Caspase-3 mRNA表达低于活血组(P<0.05)。结论活血药、破血药均能改善动脉粥样硬化,且破血药效果优于活血药;活血药可能通过抑制Cyt-C表达,降低Caspase-3 mRNA水平,破血药可能通过抑制Cyt-C表达,下调凋亡诱导因子AIF表达及降低Caspase-3 mRNA水平来抗动脉粥样硬化。

Objective To study the possible mechanism of blood-activating drug [Danggui（ Angelicaesenensis Radix 当归） and Chuanxiong（ Ligusticum Wallichii 川芎） ]and stasis-breaking drug [Sanleng（ Rhizoma Sparganii 三棱） and Ezhu（ Curcuma Zedoary 莪术） ]in improving atherosclerosis. Methods Sixty rats were randomly divided into a blank group,a model group,a statin group,a blood-activating group,and a stasis-breaking group. There were8 rats in the blank group,and 13 rats in each of the other groups. Rat model of atherosclerosis was established with high-fat diet combined with intraperitoneal injection with vitamin D3. After successfully modeling,10 ml /（ kg·d） of boiled water was given to rats in the blank group and the model group,and 1. 8 g /（ kg·d） of decoction of Danggui and Chuanxiong,decoction of Sanleng and Ezhu,or atorvastatin calcium were given to the rats in the blood-activating group,the stasis-breaking group and the statin group,respectively,once a day,for four weeks. Pathological changes of aortic tissue was observed under light microscope. Expression of cytochrome C（ Cyt-C） and the apoptosis inducing factor（ AIF） protein in aortic tissue were detected with immunohistochemical method,and the expression of cysteine aspartic protease 3（ Caspase-3） mRNA in aortic tissue was detected using in-situ hybridization method. Results Compared with the model group,atherosclerosis of aortic tissue relieved in varying degrees in the statin group,the blood-activating group and the stasis-breaking group,especially in the statin group and the stasis-breaking group.Compared with the blank group,the expression of Cyt-C,AIF protein and Caspase-3 mRNA of aortic tissue increased significantly in the model group（ P 〈0. 05,P 〈0. 01）; Compared with the model group,the expression of Cyt-C,AIF protein,and Caspase-3 mRNA in aortic tissue decreased in the statin group and the stasis-breaking group,and the expression of Cyt-C and Caspase-3 mRNA decreased in the blood-activating group（ P 〈0. 05 or P 〈0. 01）; The expression of Caspase-3 mRNA in aortic tissue in the stasis-breaking group was lower than that in the blood-activating group（ P 〈0. 05）. Conclusion Both blood-activating drug and stasis-breaking drug can improve atherosclerosis,and the effect of stasis-breaking drug is better than that of blood-activating drug; Blood-activating drug may inhibit the expression of Cyt-C and reduce the level of Caspase-3 mRNA to resist atherosclerosis. On this basis,stasis-breaking drug also may reduce the expression of apoptosis inducing factor AIF to resist atherosclerosis.