摘要
目的探讨肾小管上皮细胞来源包含miroRNA-21(miR-21)的微囊泡(microvesicles,MVs)在心肌肥大中的作用及机制。方法用转化生长因子(transforming growth factor,TGF)-β1刺激肾小管上皮细胞,采用差速离心的方法提取条件培养液中的MVs,Dil-C18染料标记肾小管上皮细胞并用荧光显微镜观察受体心肌细胞,实时荧光定量聚合酶链式反应(qPCR)检测miR-21含量,目镜测微尺测量心肌细胞长径,二喹啉甲酸(BCA)蛋白试剂盒及人心钠肽(ANP)试剂盒检测细胞蛋白含量及ANP水平。预先用miR-21 inhibitor转染受体心肌细胞,观察对细胞长径、蛋白含量及ANP水平的影响。结果TGF-β1可诱导供体肾小管上皮细胞产生MVs并进入受体心肌细胞,使心肌细胞长径、蛋白含量、ANP水平明显增加。TGF-β1诱导供体肾小管上皮细胞产生的MVs中含有miR-21,并可诱导受体心肌细胞肥大,预转染miR-21抑制剂可抑制MVs诱导的细胞肥大效应。结论肾小管上皮细胞损伤时分泌包含miR-21的MVs可诱导心肌细胞的肥大,其可能参与了慢性肾脏病并发心力衰竭的进展。
Objective To investigate the effect and mechanism of microvesicles(MVs)containing miroRNA-21(miR-21)from tubular epithelial cells in cardiomyocyte hypertrophy.Methods Renal tubular epithelial cells were stimulated by transforming growth factorβ1(TGF-β1),and the conditioned medium was extracted by differential centrifugation.Renal tubular epithelial cells were labeled with Dil-C18 dye and the recipient cardiomyocytes were observed by fluorescence microscope.MiR-21 level in MVs was detected by qPCR,and the length and diameter of cardiomyocytes were measured by eyepiece.Bicinchoninic acid(BCA)protein kit and atrial natriuretic peptide(ANP)kit were used to detect the content of cell protein and the level of ANP.MiR-21 inhibitor was transfected into recipient cardiomyocytes in advance to observe the effects on the length and diameter,protein content and ANP level of the cells.Results TGF-β1 could induce donor renal tubular epithelial cells to produce MVs which were then delivered into cardiomyocytes,and the diameter,protein content and ANP level of cardiomyocytes were significantly increased.Meanwhile,miR-21 in MVs could cause recipient cardiomyocyte hypertrophy.And pre-transfection of miR-21 inhibitors could inhibit MVs-induced cardiomyocyte hypertrophy.Conclusion The damaged renal tubular epithelial cells can secrete MVs containing miR-21 which can induce cardiomyocyte hypertrophy.And this may be involved in the progress of chronic kidney disease complicated with heart failure.
作者
狄佳
周华
李旻
徐婷
赵家璧
Di Jia;Zhou Hua;Li Min;Xu Ting;Zhao Jiabi(Department of Nephrology,the Third Affiliated Hospital of Soochow University,Changzhou 213003,China;Department of Rheumatology and Immunology,the Third Affiliated Hospital of Soochow University,Changzhou 213003,China;Department of Pathology,Changzhou Second People's Hospital,Nanjing Medical University,Changzhou 213003,China)
出处
《临床荟萃》
CAS
2020年第3期245-250,共6页
Clinical Focus
基金
常州市卫计委重大项目肾脏来源包含miroRNA-21的微囊泡在心肌肥大中的作用及机制研究(ZD201706)。
关键词
肾疾病
MICRORNA
微囊泡
心肌肥大
kidney diseases
microRNA
microvesicles
cardiomyocyte hypertrophy
