β3 肾上腺素能受体激活对心脏成纤维细胞纤维化的影响及潜在机制

[Effect and potential mechanism of β3-adrenoceptor activation on fibrosis in cardiac fibroblast cell].

作者信息

Zhang Y, Wang L, Zhang H, Ma Y T, Yang Y N, Chen B D, Ma M M, Zhu X L

机构信息

Department of Cardiology, First Affiliated Hospital, School of Medicine, Shihezi University, Shihezi 832008, China.

出版信息

Zhonghua Xin Xue Guan Bing Za Zhi. 2016 Jun 24;44(6):501-7. doi: 10.3760/cma.j.issn.0253-3758.2016.06.009.

DOI:10.3760/cma.j.issn.0253-3758.2016.06.009

PMID:27346263

Abstract

OBJECTIVE

To observe the effect of β3 adrenergic receptor (β3-AR) on fibrosis in cardiac fibroblasts(CFBs) and explore the related mechanisms.

METHODS

Neonatal CFBs were divided into negative control group (N-CFC): CFBs without any intervention; group treated with β3 adrenergic receptor agonist (AngⅡ-CFC-β3-AR BRL): CFBs treated with 10(-6) mol/L angiotensin Ⅱ(AngⅡ), 1 hour later treated with 10(-5) mol/L β3 adrenergic receptor agonist (β3-AR BRL37344); group treated with β3 adrenergic receptor antagonist (AngⅡ-CFC-β3-AR SR): CFBs treated with 10(-6) mol/L AngⅡ, 1 hour later treated with 10(-5) mol/L β3 adrenergic receptor antagonist (β3-AR SR59230A); and positive control group (AngⅡ-CFC): CFBs treated with 10(-6) mol/L AngⅡonly. Proliferation of CFBs was detected by the method of WST-1. Protein expression of β3-AR, transforming growth factor β1 receptor (TGF-β1-R), transforming growth factor β1(TGF-β1), Smad-2, phospho-Smad-2 (p-Smad-2), collagen-Ⅰ (COL-Ⅰ) and collagen-Ⅲ(COL-Ⅲ) was determined by Western blot assay.

RESULTS

(1) The proliferation of CFBs was the highest in AngⅡ-CFC-β3-AR BRL, followed by AngⅡ-CFC-β3-AR SR and AngⅡ-CFC group (all P<0.05 vs. N-CFC group). (2) The protein expression level of β3-AR, TGF-β1-R, TGF-β1 and p-Smad-2 was in the same order as proliferation of CFBs. (3) The expression level of COL-Ⅰ and COL-Ⅲ protein was also in the same order as proliferation of CFBs.

CONCLUSION

Activation of β3-AR may promote fibrosis of CFBs through the TGF-β/Smad signaling pathway and thus aggravate myocardial remodeling.

摘要

目的

观察β3肾上腺素能受体（β3-AR）对心脏成纤维细胞（CFBs）纤维化的影响并探讨其相关机制。

方法

将新生CFBs分为阴性对照组（N-CFC）：未进行任何干预的CFBs；β3肾上腺素能受体激动剂处理组（AngⅡ-CFC-β3-AR BRL）：用10（-6）mol/L血管紧张素Ⅱ（AngⅡ）处理CFBs，1小时后用10（-5）mol/Lβ3肾上腺素能受体激动剂（β3-AR BRL37344）处理；β3肾上腺素能受体拮抗剂处理组（AngⅡ-CFC-β3-AR SR）：用10（-6）mol/L AngⅡ处理CFBs，1小时后用10（-5）mol/Lβ3肾上腺素能受体拮抗剂（β3-AR SR59230A）处理；以及阳性对照组（AngⅡ-CFC）：仅用10（-6）mol/L AngⅡ处理CFBs。采用WST-1法检测CFBs的增殖情况。通过蛋白质印迹法测定β3-AR、转化生长因子β1受体（TGF-β1-R）、转化生长因子β1（TGF-β1）、Smad-2、磷酸化Smad-2（p-Smad-2）、Ⅰ型胶原（COL-Ⅰ）和Ⅲ型胶原（COL-Ⅲ）的蛋白表达。

结果

（1）CFBs增殖在AngⅡ-CFC-β3-AR BRL组最高，其次是AngⅡ-CFC-β3-AR SR组和AngⅡ-CFC组（与N-CFC组相比，均P<0.05）。（2）β3-AR、TGF-β1-R、TGF-β1和p-Smad-2的蛋白表达水平与CFBs增殖情况一致。（3）COL-Ⅰ和COL-Ⅲ蛋白表达水平也与CFBs增殖情况一致。