基于PI3K/Akt通路的冠心病痰瘀互结证大鼠发病机制研究  

Pathogenesis research of coronary heart disease with phlegm-stasis syndrome rat model based on PI3K/Akt pathway

作　　者：高杉[1] 王鹏伟 冯曼 王朔[1] 李琳[1] 潘晔[1] 刘昳佳 于春泉[1] GAO Shan;WANG Pengwei;FENG Man;WANG Shuo;LI Lin;PAN Ye;LIU Yijia;YU Chunquan(Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China)

机构地区：[1]天津中医药大学

出　　处：《天津中医药》2020年第2期209-213,共5页Tianjin Journal of Traditional Chinese Medicine

基　　金：国家重点基础研究计划（973计划）项目（2014CB542902）

摘　　要：[目的]基于磷脂酰肌醇-3激酶(PI3K)/丝氨酸/苏氨酸激酶(Akt)通路探讨冠心病痰瘀互结证大鼠模型的发病机制。[方法] Wistar雄性大鼠,随机分为空白组、假手术组、痰浊组、血瘀组、痰瘀互结组。第54天,血瘀组和痰瘀互结组结扎冠状动脉左前降支,假手术组只穿线不结扎。其中空白组、血瘀组、假手术组采用普通饲料喂养8周,痰浊组和痰瘀互结组采用高脂饲料喂养8周。采用末端脱氧核苷酸转移酶介导的缺口末端标记测定(Tunel)染色的方法观察心肌细胞凋亡情况,采用蛋白免疫印迹(Western blot)方法检测Akt、磷酸化Akt(p-Akt)、B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关的促凋亡蛋白(Bax)、半胱氨酸蛋白酶-3(Caspase-3)蛋白表达情况。[结果] Tunel染色结果显示:痰浊组、血瘀组和痰瘀互结组细胞凋亡明显增多。Western blot结果显示,与空白组比较,血瘀组和痰瘀互结组Akt、p-Akt和Bcl-2蛋白表达水平明显降低(P<0.05或P<0.01);Bax和Caspase-3蛋白表达水平明显升高(P<0.05或P<0.01);痰浊组p-Akt蛋白表达明显升高(P<0.05),Akt、Bcl-2、Bax和Caspase-3蛋白表达无统计学差异(P>0.05)。[结论]心肌细胞Akt、p-Akt和Bcl-2蛋白的下调,Bax和Caspase-3蛋白的上调,可以激活PI3K/Akt通路,促进细胞凋亡,此过程可能是冠心病痰瘀互结证的发病机制之一。[Objective] To investigate the pathogenesis of the rat model of coronary heart disease with phlegm and blood-stasis syndrome based on PI3K/Akt pathway. [Methods] Wistar male rats were randomly divided into blank group,sham operation group,turbidity group,blood stasis group and phlegm and blood-stasis group. On the 54th day,the left anterior descending coronary artery of the rats in bloodstasis group and the phlegm and blood-stasis group were ligated,and the rats in the sham operation group only threaded without ligation.The rats in the blank group,the blood-stasis group and the sham operation group were fed with normal feed for 8 weeks,while the phlegm group and the phlegm and blood-stasis group were fed with high fat diet for 8 weeks. The apoptosis of cardiomyocytes was observed by Tunel staining. The expressions of Akt,p-Akt,Bax,Bcl-2 and Caspase-3 were detected by Western blot. [Results] The results of Tunel staining showed that the apoptosis of phlegm group,blood-stasis group and phlegm and blood-stasis group increased significantly.Western blot results showed that the expression levels of Akt,p-Akt and Bcl-2 protein in blood-stasis group and phlegm and blood-stasis group were significantly lower than those in the blank group(P <0.05 or P <0.01). The expression levels of Bax and Caspase-3 were significantly increased(P<0.05 or P<0.01). The expression of p-Akt protein in the phlegm group was significantly increased(P<0.05),and there was no significant difference in the expression of Akt,Bcl-2,Bax and Caspase-3. [Conclusion] It is suggested that PI3K/Akt pathway maybe one of the pathogenesis of coronary heart disease with phlegm and blood-stasis syndrome by down-regulating Akt,p-Akt and Bcl-2 proteins in cardiomyocytes,up-regulating Bax and Caspase-3 proteins.

关 键 词：冠心病 痰瘀互结证 PI3K/AKT 细胞凋亡 动物模型 

分 类 号：R285.5[医药卫生—中药学]