摘要
目的:初步明确血浆外泌体及其内容物微小RNA-25-3p(microRNA-25-3p,miR-25-3p)对心肌纤维化的影响。方法:分别提取10只健康C57BL/6小鼠(对照组)和10只心肌梗死术后小鼠(心肌纤维化模型组)的血浆外泌体。利用电镜、纳米跟踪颗粒分析、标志物蛋白检测鉴定外泌体;实时荧光定量PCR(real-time quantitative polymerase chain reaction,q PCR)和蛋白质印迹(Western blot)检测两种外泌体对血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)诱导下的小鼠心脏成纤维细胞纤维化水平的影响;q PCR检测两种外泌体内miR-25-3p的水平;Western blot和免疫荧光染色检测miR-25-3p模拟物和抑制物对心脏成纤维细胞增殖和向肌成纤维细胞分化能力的影响。结果:对照组血浆外泌体可使AngⅡ诱导增加的纤维化相关指标下调,而心肌纤维化模型组外泌体则失去这一功能且其内miR-25-3p含量更高;miR-25-3p模拟物上调纤维化相关指标,而miR-25-3p抑制物则表现相反作用。结论:健康小鼠血浆外泌体可改善心肌纤维化;血浆外泌体内的miR-25-3p上调可促进心肌纤维化发生。
Objective:This study aims to preliminary investigate the effect of plasma exosome and the content microRNA-25-3p(miR-25-3p)on myocardial fibrosis. Methods:The plasma exosomes of 10 healthy C57BL/6 mice(control group)and 10 mice underwent myocardial infarction surgery(cardiac fibrosis model group)were collected successively. The exosomes were identified by electron microscopy,nano tracking particle analysis and detection of marker protein;q PCR and Western blot were used to investigate effects of the two different exosomes on the fibrosis level of mouse cardiac fibroblasts stimulated by angiotensin Ⅱ(Ang Ⅱ);expression of miR-25-3p in exosomes from two groups were detected by qPCR;Western blot and immunofluorescence staining were used to present the effects of miR-25-3p mimic and inhibitor on differentiation and proliferation of cardiac fibroblasts. Results:The plasma exosomes of healthy mice attenuated the increased fibrosis level induced by Ang Ⅱ,while the plasma exosomes of myocardial fibrosis model mice lost this function and contained more miR-25-3p;miR-25-3p mimic promoted cardiac fibrosis,while miR-25-3p inhibitor showed the opposite effect. Conclusion:Plasma exosome of health mice can protect myocardial fibrosis;the up-regulated miR-25-3p in plasma exosome promotes cardiac fibrosis.
作者
顾寰宇
李姗姗
王芹
姜敏
王春
GU Huanyu;LI Shanshan;WANG Qin;JIANG Min;WANG Chun(Department of Geriatrics,Nanjing Drum Tower Hospital,the Affiliated Hospital of Medical School of Nanjing University,Nanjing 210008,China)
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2022年第11期1515-1522,共8页
Journal of Nanjing Medical University(Natural Sciences)
基金
国家自然科学青年基金(81900220)
南京市卫生科技发展专项资金项目计划(YKK19052)。
