白芍总苷调控Sirt1/Foxo1通路对慢性心力衰竭大鼠的保护作用研究

Protective effect of total glucosides of paeonia regulating Sirt1/Foxo1 pathway on rats with chronic heart failure

目的:探讨白芍总苷对慢性心力衰竭大鼠的保护作用及沉默信息调节因子1(Sirt1)/叉头状转录因子1(Foxo1)通路的影响。方法:腹主动脉缩窄法建立慢性心力衰竭大鼠模型,建模成功后随机分为模型组、白芍总苷组、Sirt1激活剂组,每组8只;假手术组除不缩窄腹主动脉外,其余操作步骤相同。白芍总苷组给予1 g/kg白芍总苷胶囊生理盐水稀释液;Sirt1激活剂组给予5 mg/kg Sirt1720生理盐水稀释液;假手术组和模型组给予等体积生理盐水,连续给药28 d。无创血压测量系统读取左室舒张末期压(LVEDP)和左心室收缩压(LVSP);苏木素-伊红(HE)、Masson染色观察心肌组织形态;酶联免疫吸附(ELISA)法检测心肌组织中纤维连接蛋白(FN)、血清白细胞介素(IL)-1β、肿瘤坏死因子-α(TNF-α)水平;蛋白免疫印迹检测大鼠心肌组织中Sirt1、Foxo1、乙酰化Foxo1(Ac-Foxo1)蛋白水平。结果:模型组心肌纤维化严重,细胞破坏严重和排列紊乱,心肌细胞增生性炎症浸润现象明显;白芍总苷组、Sirt1激活剂组心肌纤维化得到缓解,细胞排列较整齐、炎症程度降低。模型组LVEDP水平、纤维化评分,心肌组织中FN水平、Ac-Foxo1蛋白水平,血清IL-1β、TNF-α水平明显高于假手术组,差异有统计学意义(q=4.661、15.058、15.834、17.689、26.087、10.157,均P<0.05);模型组LVSP水平,心肌组织中Sirt1蛋白水平明显低于假手术组,差异有统计学意义(q=9.194、18.782,均P<0.05)。与模型组相比,白芍总苷组、Sirt1激活剂组LVEDP水平、纤维化评分,心肌组织中FN水平、Ac-Foxo1蛋白水平,血清中IL-1β、TNF-α水平降低,差异有统计学意义(q=4.334、4.007、6.472、5.052、11.888、7.791、30.120、17.689、20.274、11.133、7.136、6.701,均P<0.05);而LVSP水平、心肌组织中Sirt1蛋白水平升高,差异有统计学意义(q=7.522、5.983、30.130、26.217,均P<0.05)。结论:白芍总苷对慢性心力衰竭大鼠的保护可能是通过激活Sirt1/Foxo1通路实现的。

Objective: To investigate the protective effect of total glucosides of paeonia on chronic heart failure rats and its influence on silent mating type information regulation 2 homolog 1(Sirt1)/forkhead box O transcription factor 1 (Foxo1) pathway. Methods: Rat model of chronic heart failure was established by abdominal aortic coarctation. After successful modeling,the rats were randomly divided into model group,total glucosides of paeonia group and Sirt1 activator group,with 8 rats in each group;the procedure was the same except that the abdominal aorta was not narrowed in the sham operation group. 1 g/kg total glucosides of paeonia capsule normal saline diluent was given in total glucosides of paeonia group;5 mg/kg Sirt1720 saline diluent was given in Sirt1 activator group;the same volume of saline was given in the sham operation group and the model group for 28 days consecutively. The left ventricular end diastolic pressure(LVEDP) and left ventricular systolic pressure(LVSP) were measured by noninvasive blood pressure measurement system;myocardial morphology was observed by hematoxylin eosin (HE) and Masson staining;the levels of fibronectin(FN) in myocardial tissue,interleukin (IL)-1β and tumor necrosis factor-α(TNF-α) in serum were measured by enzyme-linked immunosorbent assay(ELISA);and the protein levels of Sirt1,Foxo1 and acetylated Foxo1(Ac-Foxo1) in myocardial tissue were detected by Western blotting. Results: The myocardial fibrosis was serious,the cell destruction was serious and the arrangement was disordered,and the proliferative inflammatory infiltration of the cardiomyocytes was obvious in the model group;myocardial fibrosis was alleviated,the cells were arranged more neatly and the degree of inflammation was reduced in total glucosides of paeonia group and Sirt1 activator group. LVEDP level,fibrosis score,FN level in myocardial tissue,Ac-Foxo1 protein level,serum IL-1β and TNF-α levels in model group were significantly higher than those in sham operation group(q=4.661,15.058,15.834,17.689,26.087,10.157,all P< 0.05);LVSP level and Sirt1 protein level in myocardial tissue in model group were significantly lower than those in sham group(q=9.194, 18.782,all P< 0.05). Compared with the model group,LVEDP level, fibrosis score,FN level in myocardial tissue,Ac-Foxo1 protein level,serum IL-1β and TNF-α levels in total glucosides of paeonia group and Sirt1 activator group were significantly lower (q=4.334,4.007,6.472,5.052,11.888,7.791,30.120,17.689,20.274,7.136,6.701,all P< 0.05);and LVSP level and Sirt1 protein level in myocardium were significantly higher(q=7.522,5.983,30.130,26.217,all P< 0.05). Conclusion: The protective effect of total glucosides of paeonia on rats with chronic heart failure may be achieved by activating Sirt1/Foxo1 pathway.