参附强心丸对心肾综合征模型大鼠心肾组织自噬蛋白Beclin-1和LC3表达的影响

Effect of Shenfuqiangxin Pill on the expression of autophagy proteinsBeclin-1 and LC3 in heart and kidney tissues of rats with cardiorenal syndrome

目的:探讨参附强心丸对心肾综合征(cardiorenal syndrome,CRS)模型大鼠心、肾组织中自噬相关蛋白表达的影响,为其在治疗心肾综合征提供可能的作用机制。方法:采用腹主动脉缩窄合并肾脏急性缺血再灌注损伤制备心肾综合征大鼠模型,于术后8周随机分为5组:假手术组、模型对照组、参附强心丸高剂量组(3.24 g/kg)、低剂量组(1.62 g/kg)、雷帕霉素(RP)组(2 mg/kg),按方案剂量给药1次/d,周期为4周。待末次给药1 h后,进行麻醉后取血,检测血清中血管紧张素(ACE)、甲状腺激素(fT3)及精氨酸加压素(AVP);剖解心脏和左肾脏称重后,计算心脏指数和左肾指数;采用蛋白免疫印迹法检测心、肾组织中自噬相关基因LC3和Beclin-1蛋白的表达。结果:与假手术组比较,模型组大鼠出现ACE、fT3和AVP含量明显改变,心脏指数和肾脏指数增加的心肾共损的病理情况。通过WB检测发现,与假手术组比较,模型大鼠心脏和肾脏中LC3的表达强度均明显增强。而Beclin-1却出现心脏和肾脏的表达强度不一致的情况。说明在心肾同病的病理状态下,Beclin-1的表达调控通路不同。与模型对照组比较,参附强心丸给药组血清中ACE、fT3和AVP含量明显改善,尤以高剂量组显著(P<0.01);在心、肾组织中LC3和Beclin-1表达较模型对照组明显改善(P<0.05)。结论:参附强心丸能够改善CRS模型大鼠心肾功能,其作用机制可能与降低心肾组织中细胞自噬相关因子的表达有关。

Objective:To investigate the effect of Shenfuqiangxin(SFQX)pills in regulating the expression of autophagy-related proteins LC3 and Beclin-1 in rats with the cardiorenal syndrome(CRS)to provide a basis for further elucidation of the mechanism.Methods:A rat model of CRS was established by abdominal aortic coarctation combined with acute ischemia-reperfusion injury.Eight weeks after the operation,the rats were randomly divided into five groups:sham operation group,model control group,SFQX pills high-dose group(3.24 mg/kg),low-dose group(1.62 mg/kg),and rapamycin(RP)group(2 mg/kg).One hour after the last administration,the blood samples were taken after anesthesia to detect serum angiotensin(ACE),thyroid hormone(fT3),and arginine vasopressin(AVP).The heart and left kidney index were calculated after the heart,and left kidney was dissected and weighted.Western blot was used to detect the expression of autophagy-related genes LC3 and Beclin-1 in the heart and kidney.Results:The indexes of cardiac and renal function in the model group were significantly changed.Compared with the blank control group,LC3 expression in the heart and kidney tissues of the model control group increased(P<0.05).However,Beclin-1 expression intensity was inconsistent between heart and kidney,indicating that under the pathological state of heart and kidney disease,Beclin-1 expression regulation pathway was different.Compared with the model control group,the serum levels of ACE,fT3,and AVP in the SFQX pill group improved significantly,especially in the high-dose group(P<0.01).The expression of LC3 and Beclin-1 in heart and kidney tissues was significantly improved compared with that in the model control group.Conclusion:SSFQX pill can improve the cardiorenal function in CRS model rats.The mechanism may be related to the decrease of the expression of autophagy-related factors in the cardiorenal syndrome tissues.