3′tRF-PheGAA对AngⅡ诱导的心肌细胞肥大的调控作用及其机制  

Regulatory effect of 3′-tRF-PheGAA on AngⅡ-induced cardiomyocyte hypertrophy and its mechanism

作　　者：程雪丽 王凯[1] 赵炎 王昆[1] CHENG Xueli;WANG Kai;ZHAO Yan;WANG Kun(Institute of Translational Medicine,Qingdao University,Qingdao 266073,China)

机构地区：[1]青岛大学转化医学研究院,山东青岛266073

出　　处：《精准医学杂志》2024年第3期194-198,共5页Journal of Precision Medicine

基　　金：国家自然科学基金资助项目(82070313)。

摘　　要：目的探讨3′tRF-PheGAA对血管紧张素Ⅱ(AngⅡ)诱导的心肌细胞肥大的调控作用及其机制。方法获取小鼠乳鼠的原代心肌细胞,培养24 h以后分为A~G组。其中A组使用DMEM/F12完全培养液培养心肌细胞36 h;B组使用NC转染心肌细胞36 h;C组使用agomir-3′tRF-PheGAA转染心肌细胞36 h;D组使用DMEM/F12完全培养液培养心肌细胞84 h;E组在心肌细胞中加入终浓度为1μmol/L的AngⅡ处理48 h;F组使用anta-NC转染心肌细胞36 h,随后加入终浓度1μmol/L的AngⅡ处理48 h;G组使用anta-3′tRF-PheGAA转染心肌细胞36 h,随后加入终浓度1μmol/L的AngⅡ处理48 h。采用实时荧光定量PCR(RT-qPCR)技术检测各组心肌细胞中3′tRF-PheGAA及心房钠尿肽(ANP)、脑钠肽(BNP)、β-重链肌球蛋白(β-MHC)基因表达水平;使用罗丹明标记鬼笔环肽对各组心肌细胞中F-肌动蛋白进行染色并计算心肌细胞骨架表面积。结果RT-qPCR检测结果显示,A~C组心肌细胞中3′tRF-PheGAA及ANP、BNP、β-MHC基因水平差异具有显著性(F=137.10~1061.00,P<0.05),其中与A组相比,C组3′tRF-PheGAA及ANP、BNP、β-MHC基因水平显著上调(P<0.05);D~G组心肌细胞3′tRF-PheGAA及ANP、BNP、β-MHC基因表达水平差异有显著性(F=117.60~572.30,P<0.05),其中与E组相比,G组3′tRF-PheGAA及ANP、BNP、β-MHC基因表达水平显著降低(P<0.05)。罗丹明标记的鬼笔环肽染色结果显示,A~C组心肌细胞骨架表面积差异有显著性(F=35.06,P<0.05),其中与A组相比,C组细胞骨架表面积显著增加(P<0.05);D~G组心肌细胞骨架表面积差异有显著性(F=48.05,P<0.05),其中与E组相比,G组细胞骨架表面积显著减少(P<0.05)。结论3′tRF-PheGAA通过抑制自身表达,从而缓解AngⅡ诱导的心肌细胞肥大。Objective To investigate the regulatory effect of 3′-tRF-PheGAA on angiotensinⅡ(AngⅡ)-induced cardiomyocyte hypertrophy and its mechanism.Methods Primary cardiomyocytes of neonatal mice were obtained and cultured for 24 h,and were then divided into groups A to G.The cardiomyocytes in group A were cultured with DMEM/F12 complete culture medium for 36 h.The cardiomyocytes in group B were transfected with negative control(NC)for 36 h.The cardiomyocytes in group C were transfected with agomir-3′-tRF-PheGAA for 36 h.The cardiomyocytes in group D were cultured with DMEM/F12 complete culture medium for 84 h.The cardiomyocytes in group E were treated with AngⅡat a final concentration of 1μmol/L for 48 h.The cardiomyocytes in group F were transfected with anta-NC for 36 h and then treated with AngⅡat a final concentration of 1μmol/L for 48 h.The cardiomyocytes in group G were transfected with anta-3′-tRF-PheGAA for 36 h and then treated with AngⅡat a final concentration of 1μmol/L for 48 h.Reverse transcription-quantitative polymerase chain reaction(RT-qPCR)was used to determine the gene expression levels of 3′-tRF-PheGAA,atrial natriuretic peptide(ANP),brain natriuretic peptide(BNP),andβ-myosin heavy chain(β-MHC)in the cardiomyocytes in each group.Rhodamine-labeled phalloidin was used to stain F-actin in the cardiomyocytes in each group and calculate the skeletal surface area of cardiomyocytes.Results The RT-qPCR results showed significant differences between groups A to C in the gene expression levels of 3′-tRF-PheGAA,ANP,BNP,andβ-MHC in cardiomyocytes(F=137.10-1061.00,P<0.05).The gene expression levels of 3′-tRF-PheGAA,ANP,BNP,andβ-MHC were significantly upregulated in group C compared to group A(P<0.05).There were significant differences between groups D to G in gene expression levels of 3′-tRF-PheGAA,ANP,BNP,andβ-MHC in cardiomyocytes(F=117.60-572.30,P<0.05).The gene expression levels of 3′-tRF-PheGAA,ANP,BNP,andβ-MHC were significantly decreased in group G compared to group E(P<0.0

关 键 词：RNA 非翻译小片段 血管紧张素Ⅱ 肌细胞 心脏 心脏扩大 

分 类 号：R541[医药卫生—心血管疾病]