Kiss-10通过调控GPR54/NF-κB通路改善尿毒症毒素吲哚-3-乙酸诱导的心肌损伤  

Kiss-10 Ameliorates Myocardial Damage Induced by Uremic Toxin Indole-3-Acetic Acid by Regulating the GPR54/NF-κB Pathway

作　　者：张炜 杨海龙 谢润阳 ZHANG Wei;YANG Hailong;XIE Runyang(Cardiovascular Department,Baoji High Tech Hospital,Baoji 721000,Shaanxi,China;Interventional Vascular Department,Baoji High Tech Hospital,Baoji 721000,Shaanxi,China)

机构地区：[1]宝鸡高新医院心血管内科,陕西宝鸡721000 [2]宝鸡高新医院介入血管科,陕西宝鸡721000

出　　处：《心血管病学进展》2025年第2期179-185,共7页Advances in Cardiovascular Diseases

基　　金：陕西省卫生健康委科研课题(20220303011230)。

摘　　要：目的探究亲吻促动素-10(Kiss-10)通过调控G蛋白偶联受体54(GPR54)/核因子κB(NF-κB)通路对尿毒症毒素吲哚-3-乙酸(IAA)诱导的心肌损伤的影响机制。方法培养H9c2心肌细胞,利用CCK-8试剂盒检测心肌细胞活性筛选IAA与Kiss-10干预浓度,分为对照组、IAA组与Kiss-10组。将30只小鼠分为如上所述3组,每组10只。免疫荧光染色检测心肌细胞大小。实时荧光定量PCR检测心房利尿钠肽(ANP)、脑利尿钠肽(BNP)与心肌β-肌球蛋白重链(β-MHC)的mRNA水平。蛋白质印迹法检测GPR54与NF-κB蛋白水平。ELISA试剂盒检测白细胞介素(IL)-1β、IL-6、肿瘤坏死因子α(TNF-α)含量。苏木精-伊红染色检测心肌组织病理变化。超声心动图检测左心室后壁收缩末期厚度(LVPWs)、左心室后壁舒张末期厚度(LVPWd)、左心室前壁收缩末期厚度(LVAWs)、左心室前壁舒张末期厚度(LVAWd)及舒张早期充盈速度/心房收缩期充盈速度(E/A)。结果使用不同浓度IAA(5、10、30、50、100μmol/L)处理心肌细胞24 h,随着IAA浓度的增加,心肌细胞活性逐渐降低(P<0.05,P<0.01),其中50μmol/L约为半数心肌细胞活性抑制浓度,因此选该浓度继续后续实验。不同浓度Kiss-10(5、10、20μmol/L)对心肌细胞活性无显著性影响(P>0.05)。与对照组相比,IAA组心肌细胞活性降低(P<0.05),心肌细胞增大,ANP、BNP与β-MHC的mRNA水平升高(P<0.05),心肌细胞与心肌组织中GPR54表达减少,NF-κB表达增加(P<0.05),细胞上清液与血清中IL-1β、IL-6、TNF-α含量增加(P<0.05),小鼠心肌明显肥厚,LVPWs、LVPWd、LVAWs及LVAWd增加,E/A降低(P<0.05);与IAA组相比,Kiss-10组心肌细胞活性随Kiss-10浓度增加而逐渐升高(P<0.05),后期选择20μmol/L的Kiss-10进行研究,Kiss-10组心肌细胞变小,ANP、BNP与β-MHC的mRNA水平降低(P<0.05),心肌细胞与心肌组织中GPR54表达增加,NF-κB表达减少(P<0.05),细胞上清液与血清中IL-1β、IL-6、TNF-α含量减少(P<0.05),小鼠�Objective To investigate the effect of kisspeptin-10(Kiss-10)on myocardial injury induced by uremic toxin indole-3-acetic acid(IAA)through regulating G protein-coupled receptor 54(GPR54)/nuclear factor-κB(NF-κB)pathway.Methods H9c2 cells were cultured,and CCK-8 kit was used to detect the activity of cardiomyocytes to screen the intervention concentrations of IAA and Kiss-10.Thirty mice were divided into 3 groups as described above,with 10 mice in each group.The size of cardiomyocytes was detected by immunofluorescence staining.The mRNA levels of atrial natriuretic peptide(ANP),brain natriuretic peptide(BNP)and cardiac myosin heavy chainβ(β-MHC)were detected by real time fluorogenic quantitative PCR.Western blotting was used to detect the protein levels of GPR54 and NF-κB.The levels of interleukin(IL)-1β,IL-6 and tumor necrosis factor-α(TNF-α)were detected by ELISA kits.hematoxylin and eosin staining was used to detect the pathological changes of myocardial tissue.Left ventricular posterior wall end-systolic thickness(LVPWs),left ventricular posterior wall end-diastolic thickness(LVPWd),left ventricular anterior wall end-systolic thickness(LVAWs),left ventricular anterior wall end-diastolic thickness(LVAWd)and E/A ratio were measured by echocardiography.Results Different concentrations of IAA(5,10,30,50,and 100μmol/L)were used to treat cardiomyocytes for 24 h.With the increase of IAA concentration,the viability of cardiomyocytes was gradually decreased(P<0.05,P<0.01),and 50μmol/L was about the 50%inhibitory concentration of cell viability,so this concentration was selected for subsequent experiments.Different concentrations of Kiss-10(5,10,20μmol/L)had no significant effect on the viability of cardiomyocytes(P>0.05).Compared with the control group,the activity of cardiomyocytes was decreased(P<0.05),the cardiomyocytes were enlarged,the mRNA levels of ANP,BNP andβ-MHC were increased(P<0.05),the expression of GPR54 in cardiomyocytes and myocardial tissue was decreased,and the expression of NF-κB was inc

关 键 词：吲哚-3-乙酸 心肌损伤 亲吻促动素-10 GPR54/NF-κB通路 炎症反应 

分 类 号：R692.5[医药卫生—泌尿科学] R542.2[医药卫生—外科学]