苦碟子通过NLRP3对急性心梗大鼠心肌细胞损伤及纤维化的作用研究  

Effects of Kudiezi on cardiomyocyte injury and fibrosis in rats with acute myocardial infarction through NLRP3

作　　者：尹国婷 刘洁云 王要鑫 连政 韩文杰 Yin Guoting;Liu Jieyun;Wang Yaoxin;Lian Zheng;Han Wenjie(Department of Cardiology,Kaifeng Central Hospital,Kaifeng 475000,China;不详)

机构地区：[1]开封市中心医院心血管内科,开封475000 [2]首都医科大学附属北京同仁医院心血管中心,北京100730

出　　处：《中国循证心血管医学杂志》2023年第6期683-688,共6页Chinese Journal of Evidence-Based Cardiovascular Medicine

基　　金：河南省医学科技攻关计划(联合共建)项目(LHGJ20191067)。

摘　　要：目的探讨苦碟子注射液(简称“苦碟子”)对急性心肌梗死(AMI)大鼠心肌细胞损伤和纤维化的影响及可能作用机制。方法随机选取8只SPF级雄性Wistar大鼠作为假手术组,剩余大鼠采用左冠状动脉(冠脉)前降支结扎法建立AMI模型,成模大鼠随机分为模型组、苦碟子低剂量组(3 ml/kg·d)、苦碟子中剂量组(6 ml/kg·d)及苦碟子高剂量组(12 ml/kg·d),每组各8只,其中假手术组及模型组大鼠每日腹腔注射等量生理盐水,连续21 d。检测心功能参数;HE染色法检测心肌组织病理学损伤情况;Masson染色法检测心肌间质纤维化情况;全自动生化分析仪测定血清乳酸脱氢酶(LDH)、肌钙蛋白T(TnT)水平及肌酸激酶MB(CK-MB)活性;ELISA法检测大鼠心肌组织肿瘤坏死因子-α(TNF-α)、白细胞介素6(IL-6)、白细胞介素1β(IL-1β)含量;Western blot法检测心肌组织NOD样受体蛋白3(NLRP3)、凋亡相关斑点样蛋白(ASC)、切割天冬氨酸特异性半胱氨酸蛋白酶1(C-caspase-1)、切割IL-1β(C-IL-1β)、Ⅰ型胶原蛋白(ColⅠ)、Ⅲ型胶原蛋白(ColⅢ)、波形蛋白(Vimentin)及α-平滑肌肌动蛋白(α-SMA)蛋白表达情况。结果与假手术组比较,模型组及3个给药组大鼠左心室收缩末期直径(LVESD)、左心室舒张末期直径(LVEDD)值、血清LDH、TnT及CK-MB水平、心肌组织中TNF-α、IL-6及IL-1β含量、NLRP3、ASC、C-caspase-1及C-IL-1β蛋白相对表达量、ColⅠ、ColⅢ、Vimentin及α-SMA蛋白相对表达量升高,缩短分数(FS)及射血分数(EF)降低(P<0.05),同时可见心肌组织发生不同程度的损伤及胶原蛋白沉积;与模型组比较,苦碟子低、中、高剂量组大鼠LVESD、LVEDD值、血清LDH、TnT及CK-MB水平、心肌组织中TNF-α、IL-6及IL-1β含量、NLRP3、ASC、C-caspase-1及C-IL-1β蛋白相对表达量、ColⅠ、ColⅢ、Vimentin及α-SMA蛋白相对表达量降低,FS、EF值升高(P<0.05),心肌组织病理损伤及胶原蛋白沉积程度均减轻;苦碟子作Objective To investigate the effect of Kudiezi Injection(Kudiezi for short)on cardiomyocyte injury and fibrosis in rats with acute myocardial infarction(AMI)and its possible mechanism.Methods Eight SPF grade male Wistar rats were randomly selected as the sham operation group.The remaining rats were ligated with the anterior descending branch of the left coronary artery to establish the AMI model.The model rats were randomly divided into model group,Kudiezi low-dose group(3 ml/kg·d),Kudiezi medium dose group(6 ml/kg·d)and Kudiezi highdose group(12 ml/kg·d),with 8 rats in each group.The rats in the sham operation group and model group were intraperitoneally injected with the same amount of normal saline every day,21 consecutive days.Detect cardiac function parameters.HE staining was used to detect myocardial histopathological injury.Masson staining was used to detect myocardial interstitial fibrosis.The levels of serum lactate dehydrogenase(LDH),troponin T(TnT)and the activity of creatine kinase MB(CK-MB)were measured by automatic biochemical analyzer.Content of tumor necrosis factor-α(TNF-α),interleukin 6(IL-6),interleukin 1β(IL-1β)in rat myocardial tissue was detected by ELISA.Western blot was used to detect Nod like receptor protein 3(NLRP3),apoptosis related spot like protein(ASC),cleavage aspartate specific cysteine protease-1(C-caspase-1)and cleavage IL-1β(C-IL-1β),typeⅠcollagen(ColⅠ),typeⅢcollagen(ColⅢ),vimentin andα-Smooth muscle actin(α-SMA)protein expression.Results Compared with the sham operation group,the values of left ventricular end systolic diameter,left ventricular end diastolic diameter,the levels of serum LDH,TnT and CK-MB,and TNF-α,IL-6 and IL-1βcontent,the relative expression of NLRP3,ASC,C-caspase-1 and C-IL-1β,ColⅠ,ColⅢ,Vimentin andα-SMA protein increased in myocardial tissue in the model group and three administration groups,and the values of fractional shortening and ejection fraction decreased(P<0.05).Compared with the model group,the levels of LVESD and LVEDD,s

关 键 词：急性心肌梗死 苦碟子注射液 心肌细胞损伤 纤维化 NOD样受体蛋白3 

分 类 号：R542.22[医药卫生—心血管疾病]