甘草次酸、甘草苷配伍次乌头碱对慢性心衰大鼠细胞凋亡通路的影响

Influence of compatibility of glycyrrhetinic acid,liquiritin and hypaconitine on apoptosis pathway in rats with chronic heart failure

目的探讨甘草次酸、甘草苷配伍次乌头碱对慢性心衰大鼠的抗心衰作用及可能机制。方法 60只SD大鼠通过主动脉弓缩窄术造成慢性心衰模型,4周后随机分为假手术组(6只)、模型组(7只)、地高辛组(8只)、次乌头碱组(次乌组,9只)、甘草次酸+次乌头碱组(甘次+次乌组,9只)、甘草苷+次乌头碱组(甘苷+次乌组,9只)、甘草次酸+甘草苷+次乌头碱组(甘次+甘苷+次乌组,9只)。连续给药1周后,于第7天处死大鼠,处死前取血并行超声心动图,测得射血分数(EF)及缩短分数(FS)情况;并应用ELISA法检测血清B型钠尿肽(BNP)改变;应用免疫组化法检测心肌细胞凋亡通路Bcl-2、Bax及Caspase-3的表达情况;并应用Western blot法检测Fas、Fas-L蛋白表达情况。结果与模型组相比,除次乌组外,其它各药物配伍组的Bcl-2表达均高于模型组,同时Bax、Caspase-3、Fas与Fas-L表达及EF、FS及BNP值均低于模型组;其中以甘次+甘苷+次乌组作用效果最明显。结论甘草次酸、甘草苷配伍次乌头碱可下调EF、FS及BNP的水平及促凋亡蛋白表达,上调抑制凋亡蛋白表达,其可能是甘草配伍附子对慢性心衰的抗凋亡作用机制之一。

AIM To explore the anti-heart failure effects of compatibility of glycyrrhetinic acid,liquiritin and hypaconitine on apoptotic pathway in rats with chronic heart failure and the possible mechanism of action.METHODS Sixty rats were operated through transverse aortic constriction to create the chronic heart failure models.After four weeks,these rats were randomly divided into seven groups： sham group （n = 6）,model group （n = 7）,digoxin group （n = 8）,hypaconitine group （n = 9）,glycyrrhetinic acid ＋ hypaconitine group （n = 9）,liquiritin ＋ hypaconitine group （n = 9）,glycyrrhetinic acid ＋ liquiritin ＋ hypaconitine group （n = 9）.Then the rats were given medicines or saline perfusion by oral gavage for one week.On the 7thday,the rats were executed.Before the rats were killed,the blood samples were obtained and echocardiographic were carried on to get the ejection fraction （EF） and fractional shortening （FS） data; and the ELISA test was done for type B natriuretic peptide （BNP） changes; and myocardial histopathological examinations were performed on Bcl-2,Bax and Caspase-3.And the protein expressions of Fas and Fas-L were detected by Western blot.RESULTS Compared with the model group,the expression of Bcl-2 of all the compatibility groups except the hypaconitine group was higher than that of the model group; the expressions of Bax,Caspase-3,Fas and Fas-L,besides with the measurements of EF,FS and BNP were lower than those of the model group,and the glycyrrhetinic acid ＋ liquiritin ＋ hypaconitine group showed the best effect.CONCLUSION The compatibility of hypaconitine,glycyrrhetinic acid and liquiritin could down-regulate the levels of EF,FS and BNP,together with the protein expressions of pro-apoptosis,meanwhile,up-regulate the protein expressions of anti-apoptosis.This might be one of the mechanisms for the anti-apoptosis effects on chronic heart failure by compatibility of Glycyrrhizea radix et rhizoma （Gancao） and Acontti lateralis Radix praparata （Fuzi）.