病理状态对心肌细胞及心脏成纤维细胞β3-肾上腺素受体表达的影响

Effects of pathological conditions on the expression of β3-adrenergic receptors in cardiomyocytes and cardiac fibroblasts

目的观察与心力衰竭相关的不同病理因素刺激下,Sprague-Dawley(SD)乳鼠心肌细胞和心脏成纤维细胞中β3-肾上腺素受体(β3-AR)表达变化的差异。方法分离培养SD乳鼠心肌细胞和心脏成纤维细胞并行免疫荧光鉴定,分别给予血管紧张素Ⅱ(AngⅡ)及去甲肾上腺素(NE)进行刺激,采用实时荧光定量PCR(qPCR)检测心肌细胞肥大相关基因[心钠肽(ANP)、脑钠肽(BNP)、β-肌球蛋白重链(β-MHC)]、心脏成纤维细胞纤维化相关基因[Ⅰ型胶原蛋白(COL-Ⅰ)、Ⅲ型胶原蛋白(COL-Ⅲ)]及2种细胞中β3-AR的mRNA表达。结果免疫荧光鉴定显示,分离培养的心肌细胞和心脏成纤维细胞均状态良好。AngⅡ和NE分别刺激2种细胞48h后,心肌细胞中ANP、BNP、β-MHC和β3-AR的表达较对照组明显增加;心脏成纤维细胞中COL-Ⅰ、COL-Ⅲ表达较对照组明显增加,但β3-AR的表达没有明显变化。结论参与心力衰竭发生发展的病理因素AngⅡ和NE主要引起心肌细胞的β3-AR表达增加,而不是心脏成纤维细胞。

Objective To observe differences in the expression of β3-adrenergic receptor (β3-AR) under different pathological factors associated with heart failure in neonatal rat cardiac myocytes and cardiac fibroblasts. Methods The neonatal rat cardiomyocytes and cardiac fibroblasts were isolated and cultured, and both were identified by immunofluorescence. Then angiotensin Ⅱ(AngⅡ) and norepinephrine (NE) were given to stimulate the cells. And qPCR was used to detect cardiomyocyte hypertrophy-related genes (ANP, BNP and β-MHC), cardiac fibroblast-related genes (Col-Ⅰ, COL-Ⅲ) and β3-AR mRNA expression in two kinds of cells. Results Immunofluorescence identification showed that the isolated and cultured cardiomyocytes and cardiac fibroblasts were in good condition. After 48 hours of AngⅡ and NE stimulation, the expressions of ANP, BNP,β-MHC and β3-AR were increased significantly in cardiomyocytes compared with those in control group, while the expressions of Col-Ⅰ and COL-Ⅲ were increased significantly in cardiac fibroblasts compared with those of the normal control group. However, the expression of β3-AR did not change significantly. Conclusion AngⅡ and NE, pathological factors involved in the development of heart failure, mainly cause an increase in the expression of β3-AR in cardiomyocytes rather than cardiac fibroblasts.