真武汤对心力衰竭大鼠心肌细胞凋亡及PI3K-AKT通路的影响

Effect of Zhenwu decoction on cardiomyocyte apoptosis and the PIth congestive heart failure3K-AKT pathway in rats wi

目的观察真武汤对冠脉结扎心衰大鼠模型的治疗作用及对心衰大鼠心肌病理形态学及心肌细胞凋亡的影响,探讨其对凋亡相关蛋白及PI3K-AKT信号通路的影响。方法术后4周采用随机数字表法将存活的大鼠分为6组,每组7只:(1)假手术组:术后4周给予生理盐水灌胃28 d;(2)模型组:造模4周后予生理盐水灌胃28 d;(3)阳性对照组:造模4周后予氯沙坦钾5 mg/kg灌胃28 d;(4)真武汤低剂量组:造模4周后予真武汤生药量6 g/kg灌胃28 d;(5)真武汤中剂量组:造模4周后予真武汤生药量12 g/kg灌胃28 d;(6)真武汤高剂量组:造模4周后予真武汤生药量18 g/kg灌胃28 d。观察各组大鼠造模前后心电图、彩超变化、心肌细胞凋亡、心肌形态学变化、心肌组织PI3K、AKT1、p-AKT1蛋白水平变化、免疫组化Bax、Bcl-2、Caspase3、8、9变化及腹主动脉血BNP变化。结果(1)心电图显示经过冠脉结扎心衰模型制备大鼠心电图ST段抬高,提示造模成功;(2)心衰大鼠左心质量指数、BNP水平显著升高,彩超提示LVEF、LVFS均明显下降,其余各组左心质量指数、BNP水平、LVEF、LVFS均有改善(P<0.01);(3)HE染色提示模型组心肌纤维排列紊乱、断裂,同时并伴随着心肌细胞肿胀、肥大,有炎性细胞浸润,其余各组存在不同程度的缓解。TUNEL染色提示模型组凋亡现象严重,经真武汤及阳性药干预后凋亡现象减轻;(4)免疫组化结果提示与模型组相比,各给药组大鼠Bcl-2升高,Bax、Caspase3、8、9下降,其中真武汤中剂量组各蛋白表达与假手术无明显差异;(5)与假手术组相比,模型组PI3K、p-AKT1、AKT1表达均升高,真武汤可抑制PI3K、p-AKT1、AKT1蛋白量的表达(P<0.05)。结论真武汤可能通过调控PI3K-AKT通路减轻HF大鼠心肌细胞凋亡及心肌病理改变,增强HF大鼠心肌收缩力,进而延缓大鼠心衰进程。

Objective To observe the therapeutic effect of Zhenwu decoction on myocardial pathomorphology and cardiomyocyte apoptosis in rats with heart failure induced by coronary artery ligation, and to explore its effect on apoptosis-related proteins and the PI3 K-AKT signaling pathway. Methods At 4 weeks after the operation, surviving rats were randomly divided into six groups with seven rats in each group as follows.(1)Sham operation group: normal saline was administered intragastrically for 28 days at 4 weeks after the operation.(2) model group: normal saline was administered intragastrically for 28 days after 4 weeks.(3) positive control group: 5 mg/kg losartan potassium was administered for 28 days after 4 weeks.(4) Zhenwu decoction low dose group: after 4 weeks, 6 g/kg Zhenwu decoction crude drug was administered for 28 days.(5) Zhenwu decoction middle dose group: 12 g/kg Zhenwu decoction crude drug was administered intragastrically for 28 days after 4 weeks.(6) Zhenwu decoction high dose group: 18 g/kg Zhenwu decoction crude drug was administered intragastrically for 28 days after 4 weeks. Changes in the electrocardiogram, color ultrasound, cardiomyocyte apoptosis, myocardial morphology, PI3 K, AKT1 and p-AKT1 protein levels in myocardial tissue, immunohistochemical changes of Bax, Bcl-2, caspase 3, 8, 9 and BNP in the abdominal aorta were observed before and after modeling. Results(1)The ST segment of the heart failure model was elevated after coronary artery ligation, indicating that model establishment was successful.(2)The left ventricular mass index and BNP level of heart failure rats were increased significantly, and LVEF and LVFS were decreased significantly, whereas the left the ventricular mass index, BNP level, LVEF and LVFS were improved in the other groups.(3)Myocardial fibers in the model group were disordered and broken accompanied by cardiomyocyte swelling, hypertrophy. and inflammatory cell infiltration, which were relieved to various degrees in the other groups. Apoptosis in the model group was significant, which was alleviated after by Zhenwu decoction and positive control group drugs.(4)Compared with the model group, Bcl-2 was increased, and Bax and Caspase 3, 8, 9 were decreased in each treatment group. There was no significant difference in protein expression between Zhenwu decoction middle dose and sham operation groups.(5)Compared with the sham operation group, PI3 K, p-AKT1 and AKT1 expression in the model group was increased, and Zhenwu decoction suppressed protein expression of PI3 K, p-AKT1 and AKT1. Conclusions Zhenwu decoction reduces cardiomyocyte apoptosis and myocardial pathological changes in HF rats by regulating the PI3 K-AKT pathway, enhancing myocardial contractility, and delaying the progression of heart failure.