单酯型生物碱类对CHF大鼠心肌细胞的保护作用及机制

Study on the Protective Effect and Mechanism of Monoester Alkaloids on Cardiomyocytes of CHF Rats

目的 分析单酯型生物碱类对慢性心力衰竭大鼠心肌细胞的保护作用及对PI3K/AKT通路的影响。方法 将100只大鼠随机分为空白组(A组)、模型组(B组)、低剂量组(C组)、中剂量组(D组)和高剂量组(E组),每组20只。C、D、E组在慢性心力衰竭模型建模成功后给予附子浓缩液灌胃,剂量分别为2.79、5.57、11.14 g/kg。灌胃6周后检测各组大鼠病理组织学变化、超声心动图指标、心肌细胞凋亡率,心肌组织促凋亡蛋白(Bax)、B淋巴细胞瘤-2(Bcl-2)、半胱氨酸蛋白酶8(Caspase-8)表达量,心肌细胞自噬蛋白、BNP、AI、PO_(2)、IL-6、TNF-α水平及PI3K/AKT通路蛋白表达量。结果 B~E组超声心动图指标室间隔收缩末期厚度(IVSs)、室间隔舒张末期厚度(IVSd)、左心室后壁收缩末期厚度(LVPWs)、左心室后壁舒张末期厚度(LVPWd)。心肌细胞凋亡率,Bax、Caspase-8表达量,IL-6、TNF-α水平,心肌细胞自噬蛋白LC3-Ⅱ、LC3-Ⅱ/Ⅰ,BNP及AI水平均高于A组,LVEF、Bcl-2表达量、PO_(2)水平、心肌细胞自噬蛋白P62及p-PI3K、p-AKT表达量均低于A组(P<0.05);C~E组IVSs、IVSd、LVPWs、LVPWd、心肌细胞凋亡率及Bax、Caspase-8表达量,IL-6、TNF-α水平、自噬蛋白LC3-Ⅱ、LC3-Ⅱ/Ⅰ、BNP及AI水平低于B组,LVEF、Bcl-2表达量、PO_(2)水平、心肌细胞自噬蛋白P62及p-PI3K、p-AKT表达量高于B组(P<0.05);随着附子浓缩液剂量的增加,IVSs、IVSd、LVPWs、LVPWd、心肌细胞凋亡率、Bax、Caspase-8表达,IL-6、TNF-α水平、心肌细胞自噬蛋白LC3-Ⅱ、LC3-Ⅱ/Ⅰ、BNP及AI水平降低,LVEF及Bcl-2表达、PO_(2)水平、心肌细胞自噬蛋白P62及p-PI3K、p-AKT表达量升高(P<0.05)。结论 单酯型生物碱类通过上调p-PI3K、p-AKT蛋白表达量,激活PI3K/AKT通路,缓解心力衰竭恶性进展,且呈剂量依赖性。

Objective To analyze the protective effects of single ester alkaloids on myocardial cells of chronic heart failure rats and their effects on PI3K/AKT pathway.Method 100 rats were randomly divided into group A(blank group),group B(model group),group C(low-dose group),group D(medium-dose group)and group E(high-dose group)with 20 rats in each group.Groups C,D and E were given aconite concentrate by intragastric administration after successful modeling of chronic heart failure model,and the doses were 2.79,5.57 and 11.14 g/kg,respectively.After 6 weeks of gavage,histopathological changes,echocardiographic examination indexes,myocardial cell apoptosis rate,expression of cardiac tissue proapoptotic protein(Bax),B-lymphoblastoma-2(Bcl-2),Cagpase-8,autophagy protein,BNP,AI of myocardial cells in each group were observed.Result The echocardiography indexes IVSs,IVSd,LVPWs,LVPWd,the apoptosis rate of cardiomyocytes,the expression of Bax,Caspase-8,the levels of IL-6,TNF-α,the expression of autophagy protein LC3-Ⅱ,LC3-Ⅱ/Ⅰ,BNP and AI in cardiomyocytes in groups E,D and C were higher than those in group A.LVEF,expressions of Bcl-2,PO_(2),autophagy protein P62,p-PI3K and p-AKT in groups E,D and C were lower than those in group A(P<0.05).The IVSs,IVSd,LVPWs,LVPWd,the apoptosis rate of myocardial cells,the expression of Bax and Caspase-8,levels of IL-6,TNF-α,autophagy protein LC3-Ⅱ,LC3-Ⅱ/Ⅰ,BNP and AI in groups E,D and C were lower than those in group B.LVEF,the expressions of Bcl-2,PO_(2),autophagy protein P62,P-PI3K and P-Akt in groups E,D and C were higher than those in group B(P<0.05).With the increase of concentration of monkshood concentrate,IVSs,IVSd,LVPWs,LVPWd,myocardial cell apoptosis rate,Bax,Caspase-8 expression,IL-6,TNF-αlevels,myocardial autophagy protein LC3-Ⅱ,LC3-Ⅱ/Ⅰ,BNP and AI levels decreased.LVEF,the expressions of Bcl-2,PO_(2),autophagy protein P62,p-PI3K and p-AKT in myocardial cells were increased(P<0.05).Conclusion Single ester alkaloids may activate PI3K/AKT pathway by up-regulating the expression of p-PI3K and p-AKT,and alleviate the malignant progression of heart failure in a dose-dependent manner.