Calhex231通过细胞焦亡改善大鼠心肌梗死面积及心肌纤维化  

Calhex231 Attenuates Rat Myocardial Infarction Size and Fibrosis by Suppressing Pyroptosis

作　　者：刘文秀[1] 郭雨桐 孙雪[1] 宋琳琳 刘越[1] 丁雪[1] LIU Wenxiu;GUO Yutong;SUN Xue;SONG Linlin;LIU Yue;DING Xue(Department of Cardiology,The First Affiliated Hospital of Harbin Medical University,Harbin 150001,Heilongjiang,China)

机构地区：[1]哈尔滨医科大学附属第一医院心内科,黑龙江哈尔滨150001

出　　处：《心血管病学进展》2024年第4期379-384,共6页Advances in Cardiovascular Diseases

基　　金：国家自然科学基金(81700318);黑龙江省自然科学基金(LH2023H022);黑龙江省博士后科研启动金(LBH-Q21109)

摘　　要：目的研究钙敏感受体(CaSR)负性变构调节剂Calhex231是否能改善大鼠的心肌梗死(MI)面积和心肌纤维化。方法健康Wistar大鼠随机分为3组:sham组、MI组和MI+Calhex231组。TTC染色评估MI面积和坏死情况。Masson染色、免疫组织化学及电镜检查心肌纤维化、Ⅲ型胶原蛋白及成纤维细胞胶原合成情况。免疫荧光和Western blot检测CaSR表达变化。Western blot检测含NOD样受体热蛋白结构域相关蛋白3(NLRP3)、胱天蛋白酶-1(Casp-1)、gasdermin D(GSDMD)、GSDMD N-末端结构域(NT-GSDMD)和白细胞介素-1β(IL-1β)的蛋白表达变化。并采用免疫组织化学法评估NT-GSDMD和IL-1β的表达情况。结果与sham组相比,MI组大鼠MI面积和纤维化明显增加,心肌组织Ⅲ型胶原蛋白沉积和成纤维细胞胶原合成明显增加,而且CaSR、NLRP3、Casp-1、GSDMD、NT-GSDMD和IL-1β表达水平也明显升高。进一步免疫组织化学染色确认了NT-GSDMD和IL-1β表达增加。MI+Calhex231组与MI组相比较,Calhex231可抑制上述改变。结论Calhex231可通过CaSR抑制细胞焦亡和Ⅲ型胶原蛋白沉积,改善大鼠MI面积和心肌纤维化,有助于Calhex231在心肌纤维化疾病中临床转化。Objective Calcium-sensing receptor(CaSR)plays an important role in the development of myocardial infarction(MI).Thus,we investigated whether Calhex231,a negative allosteric modulator of CaSR,could protect the myocardium in a rat MI model.Methods Wistar rats were randomly divided into three groups:sham,MI and MI+Calhex231.Myocardial infarct size and necrosis were assessed by TTC staining.Myocardial fibrosis,collagenⅢand collagen synthesis of fibroblasts were examined by Masson staining,immunohistochemistry and electron microscopy.CaSR expression was detected by both immunofluorescence and Western blot analysis.Protein expression levels of the NOD-like receptor thermal protein domain associated protein 3(NLRP3),caspase-1(Casp-1),gasdermin D(GSDMD),N-terminal domain of GSDMD(NT-GSDMD)and interleukin-1β(IL-1β)were detected by Western blot analysis.NT-GSDMD and IL-1βwere also assessed by immunohistochemistry.Results Compared with those of the sham group,myocardial infarct size and fibrosis were obviously increased in the rats of the MI group.CollagenⅢdeposition and collagen synthesis of fibroblasts were evidently increased in myocardial tissues of the MI group compared with the sham group.The expression levels of CaSR,NLRP3,Casp-1,GSDMD,NT-GSDMD and IL-1βwere significantly increased in the MI group compared with the sham group.The increased expression of NT-GSDMD and IL-1βwas further confirmed by immunohistochemical staining.Moreover,Calhex231 administration inhibited all these alterations in the MI+Calhex231 group compared to the MI group.Conclusion Calhex231 reduced myocardial infarct size and fibrosis in rats,probably by inhibiting pyroptosis and collagenⅢdeposition via CaSR,which contributes to the potential clinical application of Calhex231 in myocardial fibrotic disease.

关 键 词：钙敏感受体 细胞焦亡 心肌梗死 纤维化 

分 类 号：R54[医药卫生—心血管疾病]