目的 观察苓桂气化方对射血分数保留心力衰竭(heart failure with preserved ejection fraction,HFpEF)早期模型大鼠心脏舒张功能的干预效应,基于核苷酸结合寡聚化结构域样受体 3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)/胱氨酸天冬氨酸特异性蛋白酶-1(cysteine-containing aspartate-specific proteases-1,Caspase-1)/消皮素D(Gasdermin D,GSDMD)通路探讨其潜在的分子作用机制.方法 40 只自发性高血压大鼠高盐高脂高糖饮食联合腹腔注射链脲佐菌素溶液建立HFpEF早期大鼠模型,分为HFpEF组、恩格列净组(1.8 mg/kg)、苓桂气化方低剂量组(4.96 g/kg)、苓桂气化方高剂量组(9.92 g/kg),予相应药物灌胃;正常血压组、空白组予等剂量生理盐水灌胃,连续干预 9 周.采用超声心动图检测大鼠左心房内径(left atrial diameter,LAD)、舒张早期二尖瓣血流速度(left ventricular early diastolic filling peak velocity,E)与舒张晚期二尖瓣血流速度(atrial systolic left ventricular filling peak velocity,A)比值、左室射血分数(left ventricular ejection fraction,LVEF);酶联免疫吸附测定法检测血清心房钠尿肽(atrial natriuretic peptide,ANP)含量;苏木精—伊红染色观察心肌组织病理变化;蛋白质印迹(Western Blot,WB)法检测NLRP3、Caspase-1、GSDMD和白介素 1β(interleukin-1β,IL-1β)的表达.结果 与正常血压组相比,HFpEF组LAD和E/A增加(P<0.05),LVEF无明显变化(P>0.05);血清ANP含量升高(P<0.05);病理观察显示心肌炎症浸润;WB结果示NLRP3、Caspase-1、GSDMD和IL-1β蛋白含量升高(P<0.05),表明建模成功.与HFpEF组比,恩格列净组和苓桂气化方低、高剂量组的LAD和E/A减小(P<0.05),血清ANP含量降低(P<0.05),病理观察显示心肌炎症浸润减轻,WB结果示恩格列净组NLRP3、Caspase-1、GSDMD、IL-1β蛋白含量降低(P<0.05),苓桂气化方低剂量组NLRP3、IL-1β蛋白含量降低(P<0.05),苓桂气化方高剂量组NLRP3、GSDMD、IL-1β蛋白含量降低(P<0.05),其余蛋白含量有下降趋势.结论 苓桂气化方能改善 HFpEF 早期模型大鼠心脏舒张功能,其机制可能与调控NLRP3/Caspase-1/GSDMD信号通路、减轻心肌炎症浸润、抑制心房重构相关.
Exploring the molecular mechanism of Linggui Qihua Formula in improving diastolic function of early stage of heart failure with preserved ejection fraction based on NLRP3/Caspase-1/GSDMD pathway
Objective To observe the intervention effect of Linggui Qihua(LGQH)Formula on cardiac diastolic function of rats with early stage of heart failure with preserved ejection fraction(HFpEF),and explore its potential molecular mechanism based on NLRP3/caspase-1/GSDMD pathway.Methods Rat model of HFpEF was established in forty spontaneous hypertension rats by high-salt-fat-sugar diet combined with intraperitoneal injection of streptozotocin solution.The rats were divided into HFpEF group,enagliflozin group(1.8 mg/kg),low-dose LGQH(LGQH-L)(4.96 g/kg)and high-dose LGQH(LGQH-H)group(9.92 g/kg),which were given corresponding drugs by gavage.Rats in the normal blood pressure group and the blank group were given equal doses of normal saline for 9 weeks.Echocardiography was used to detect the left atrial diameter(LAD),E/A and left ventricular ejection fraction(LVEF).The method of Enzyme-linked immunosorbent assay was used to detect the levels of serum atrial natriuretic peptide(ANP).Hematoxylin-eosin staining was used to observe the pathological changes of myocardial tissue.Western blot(WB)was used to detect the expression of NOD-like receptor thermal protein domain associated protein 3(NLRP3),cystine containing aspartate specific proteins-1(Caspase-1),Gasdermin D(GSDMD)and interleukin-1β(IL-1β).Results Compared with the normal blood pressure group,the levels of LAD and E/A were increased in the HFpEF group(P<0.05),with no significant change in LVEF(P>0.05).The levels of ANP in serum were increased(P<0.05).Pathological observation revealed inflammatory infiltration of the myocardium;the results of WB showed that the expressions of NLRP3 Caspase-1,GSDMD and IL-1β were increased(P<0.05),indicated that the modeling was successful.Compared with the HFpEF group,the levels of LAD and E/A were decreased in the enagliflozin group,the LGQH-L and LGQH-H group(P<0.05).The levels of ANP in serum were decreased(P<0.05).Pathological observation showed attenuation of the inflammatory infiltrate in the myo-cardium;WB results showed that the expressions of NLRP3,Caspase-1,GSDMD and IL-1β were decreased in the enagliflozin group(P<0.05),the expression of NLRP3 and IL-1β in the LGQH-L group(P<0.05)were decreased,the expressions of NLRP3,GSDMD and IL-1β in the LGQH-H group(P<0.05)were decreased,and other expressions have a decreased trend.Conclusion LGQH formula can improve the cardiac diastolic function in early stage of HFpEF rats,and its mechanism may be related to regulating the NLRP3/Caspase-1/GSDMD signaling pathway,attenuating the inflammatory infiltration of the myocardium,and inhibiting atrial remodeling.
heart failure with preserved ejection fractionpreclinical heart failureearly stage of HFpEFNLRP3/Caspase-1/GSDMD signaling pathwayLinggui Qihua Formula
NETL
NSTL
万方数据
