基于NLRP3/Caspase-1/GSDMD信号通路探讨苓桂气化方改善射血分数保留心力衰竭早期舒张功能的分子机制  

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

作　　者：杨晨光 石玉姣 乔文博 刘永成 刘思雨 梁小雨 李知轩 张贺[1,4] 董国菊 YANG Chenguang;SHI Yujiao;QIAO Wenbo;LIU Yongcheng;LIU Siyu;LIANG Xiaoyu;LI Zhixuan;ZHANG He;DONG Guoju(First Department of Cardiology of Xiyuan Hospital of China Academy of Chinese Medical Sciences,Beijing 100091,China)

机构地区：[1]中国中医科学院西苑医院心血管一科,北京100091 [2]中国中医科学院西苑医院超声科,北京100091 [3]中国中医科学院研究生院 [4]国家中医临床心血管病医学研究中心

出　　处：《环球中医药》2024年第6期999-1006,共8页Global Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(82074423);中国中医科学院西苑医院提升中医药临床循证证据级别研究专项(XYZX0201-02);中国中医科学院科技创新重大攻关项目(CI2021A00903)。

摘　　要：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-1βwere 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 myocardium;WB results showed that th

关 键 词：射血分数保留心力衰竭 前临床心衰 射血分数保留心力衰竭早期 核苷酸结合寡聚化结构域样受体3/胱氨酸天冬氨酸特异性蛋白酶-1/消皮素D信号通路 苓桂气化方 

分 类 号：R285.5[医药卫生—中药学]