本文選題：核糖核酸類 + 腫瘤抑制蛋白質(zhì)類�。� 參考：《中國(guó)循環(huán)雜志》2017年02期

【摘要】：目的:通過(guò)下調(diào)肥大心肌細(xì)胞中微小核糖核酸(miR)-31的表達(dá),觀察miR-31對(duì)大腫瘤抑制因子2(LATS2)及心肌細(xì)胞肥大的調(diào)控作用。方法:體外分離大鼠心肌細(xì)胞并連續(xù)培養(yǎng)10天,按干預(yù)條件不同將心肌細(xì)胞分為4組:空白對(duì)照組、單純血管緊張素Ⅱ(AngⅡ)干預(yù)組、miR-31抑制物慢病毒感染組、陰性病毒感染組。培養(yǎng)第8天實(shí)時(shí)熒光定量逆轉(zhuǎn)錄聚合酶鏈?zhǔn)椒磻?yīng)(qRT-PCR)檢測(cè)各組心肌細(xì)胞miR-31、LATS2及肥大基因心房鈉尿肽(ANP)、β-肌球蛋白重鏈(β-MHC)表達(dá)。培養(yǎng)第10天心肌細(xì)胞熒光探針染色觀察心肌細(xì)胞形態(tài)變化。蛋白免疫印跡(Western blot)檢測(cè)LATS2蛋白表達(dá)變化。雙熒光素酶報(bào)告基因質(zhì)粒轉(zhuǎn)染293T細(xì)胞并檢測(cè)熒光素酶活性,鑒定miR-3l對(duì)LATS2的靶向作用。結(jié)果:與空白對(duì)照組相比,單純AngⅡ干預(yù)組miR-31、心肌肥大基因ANP、β-MHC表達(dá)水平均顯著上升(P0.05),心肌細(xì)胞相對(duì)表面積明顯增大(P0.05),而LATS2基因及蛋白表達(dá)明顯下調(diào)(P0.05);與單純AngⅡ干預(yù)組比較,miR-31抑制物慢病毒感染組miR-31、心肌肥大基因ANP、β-MHC表達(dá)明顯下調(diào)(P0.05),心肌細(xì)胞相對(duì)表面積減小(P0.05),而LATS2在基因水平略有上升,蛋白水平則顯著上調(diào)(P0.05)。雙熒光素酶報(bào)告基因檢測(cè)顯示TRAF6-3'UTR+miR-146b相對(duì)熒光素酶活性較TRAF6-3'UTR+miR-NC顯著性下降(P0.01),LATS2-3'UTR+miR-31相對(duì)熒光素酶活性較LATS2-3'UTR-NC+miR-31顯著性降低(P0.01),差異均有統(tǒng)計(jì)學(xué)意義。結(jié)論:下調(diào)肥大心肌細(xì)胞miR-31表達(dá)水平可一定程度逆轉(zhuǎn)心肌細(xì)胞肥大,miR-31靶向作用LATS2參與調(diào)控心肌細(xì)胞的肥大。
[Abstract]:Aim: to investigate the regulatory effect of miR-31 on the expression of miRN-31 in hypertrophic cardiomyocytes and the role of miR-31 in the regulation of large tumor suppressor 2 LATS2) and cardiomyocyte hypertrophy. Methods: rat cardiomyocytes were isolated in vitro and cultured for 10 days. Myocardial cells were divided into 4 groups according to different intervention conditions: blank control group, single angiotensin 鈪，

本文編號(hào)：1896195