基于ROS介导的NLRP3炎症小体活化探讨燧心汤改善心力衰竭大鼠心室重构的机制研究  

Study on mechanism of Suixin Decoction improving ventricular remodeling in heart failure rats through ROS-mediated activation of NLRP3 inflammasome

作　　者：郭丹丹[1,2] 杨振宇 季雅楠 于思明[3] 孙志刚 袁星星[1,4] 李书霖 李竹英[3] GUO Dandan;YANG Zhenyu;JI Yanan;YU Siming;SUN Zhigang;YUAN Xingxing;LI Shulin;LIZhuying(Heilongjiang University of Chinese Medicine,Harbin 150040,China;Second Affiliated Hospital,Heilongjiang University of Chinese Medicine,Harbin 150001,China;First Affiliated Hospital,Heilongjiang University of Chinese Medicine,Harbin 150040,China;Heilongjiang Academy of Traditional Chinese Medicine,Harbin 150006,China)

机构地区：[1]黑龙江中医药大学,哈尔滨150040 [2]黑龙江中医药大学附属第二医院,哈尔滨150001 [3]黑龙江中医药大学附属第一医院,哈尔滨150040 [4]黑龙江省中医药科学院,哈尔滨150006

出　　处：《中华中医药杂志》2024年第5期2281-2286,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：黑龙江省中医药科研项目(No.ZHY2020-148);黑龙江省中医药科研项目(No.ZHY2023-174);黑龙江省哈尔滨市科技局应用技术研究与开发项目(No.2017RAQXJ208)。

摘　　要：目的:观察燧心汤对活性氧(ROS)介导的NOD样受体热蛋白结构域相关蛋白3(NLRP3)炎症小体的影响,从而阐明燧心汤改善心力衰竭心室重构的分子机制。方法:SD大鼠随机分为假手术组,模型组,燧心汤低、中、高剂量组和西药组,除假手术组外,采用左前降支结扎法构建心力衰竭大鼠模型,并分别给予对应药物处理。采用超声心动图测定心功能和心室重构的指标,采用ELISA法检测各组血清肌酸激酶(CK)、CK同功酶MB(CK-MB)、乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)、丙二醛(MDA)和谷光甘肽过氧化酶(GSH-Px)的含量与活性,使用HE和Masson染色评估心肌病理形态和胶原沉积,采用Tunel染色法检测心肌细胞凋亡水平。另取H9c2心肌细胞,分为空白组,模型组,燧心汤低、中、高剂量组和西药组。除空白组外,采用缺氧/再氧化法构建细胞模型并给予相应的血清进行干预。分别采用光镜下观察心肌细胞形态,采用Tunel染色法检测心肌细胞凋亡水平,流式细胞术检测心肌细胞的ROS水平,Western Blot检测NLRP3炎症小体相关蛋白的表达。结果:与假手术组比较,模型组大鼠LVEDd、LVESd、CK、CK-MB、LDH、GSH-Px的水平和SOD的活力显著升高,心肌病理评分、胶原染色面积和Tunel阳性染色细胞数显著增加,LVEF、LVFS和MDA的水平显著降低(P<0.01)。与模型组比较,燧心汤组大鼠LVEDd、LVESd、CK、CK-MB、LDH、GSH-Px的水平和SOD的活力显著降低,心肌病理评分、胶原染色面积和Tunel阳性染色细胞数显著减少,LVEF、LVFS和MDA的水平显著增加(P<0.05,P<0.01)。此外,与空白组细胞比较,模型组Tunel阳性染色细胞数和ROS水平显著增加,NLRP3、Caspase-1 p20、Mature IL-1β、Pro IL-1β、Mature IL-18、Pro IL-18、GSDMD-NT蛋白的表达显著上调,GSDMD-FL蛋白的表达显著下调(P<0.01);与模型组比较,燧心汤组Tunel阳性染色细胞数和ROS水平显著降低,NLRP3、Caspase-1 p20、Mature IL-1β、Pro of Traditional Chinese Medicine;Objective:To observe the effect of Suixin Decoction on ROS-mediated NLRP3 inflammasome activation,to elucidate the molecular mechanism of f Suixin Decoction in improving ventricular remodeling in heart failure.Methods:SD rats were randomly divided into the sham-operation group,the model group,low-dose group,the middle-dose group,the high-dose group of Suixin Decoction,and the Western medicine group,except for the sham-operation group,the left anterior descending branch ligation method was used to construct a rat heart failure model,and the corresponding drug treatments were given respectively.Echocardiography was used to determine the level of cardiac function and ventricular remodeling indicators,ELISA was used to detect the content and activity of serum CK,CK-MB,LDH,SOD,MDA and GSH-Px in each group,HE and Masson staining were used to assess myocardial pathologic morphology and collagen deposition,and the level of apoptosis of cardiomyocytes was detected by Tunel staining.H9c2 cardiomyocytes were taken and divided into control group,model group,low-dose group,the middle-dose group,the high-dose group of Suixin Decoction and Western medicine group.Except for the control group,the cell model was constructed by hypoxia/re-oxidation(H/R)method and the corresponding serum was given for intervention.The morphology of cardiomyocytes was observed under microscopy,the apoptosis level of cardiomyocytes was detected by Tunel staining,the ROS level of cardiomyocytes was detected by flow cytometry,and the expression of NLRP3inflammasome-associated proteins was detected by Western Blot,respectively.Results:Compared with the sham-operated group,the levels of LVEDd,LVESd,CK,CK-MB,LDH,GSH-Px and the activity of SOD in the model group of rats were significantly higher,and the myocardial pathology scores,collagen staining area and the number of Tunel-positively stained cells were significantly increased,while the levels of LVEF,LVFS and MDA were significantly reduced(P<0.01).Compared with the model

关 键 词：燧心汤 心力衰竭 心室重塑 NOD样受体热蛋白结构域相关蛋白3炎症小体 机制 

分 类 号：R285.5[医药卫生—中药学]