Role of runt-related transcription factor 3 in myocardial fibrosis in mice

Qian ZHAO; Yong ZOU; Bo LIU; Xinqiang YU; Suchuan ZHANG

doi:10.16016/j.1000-5404.202106071

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Role of runt-related transcription factor 3 in myocardial fibrosis in mice

Basic Medicine | 更新时间：2026-01-09

- Role of runt-related transcription factor 3 in myocardial fibrosis in mice
- Journal of Third Military Medical University Vol. 43, Issue 24, Pages: 2640-2647(2021)
- 作者机构：
  
  430015 武汉，江汉大学附属医院心血管内科
- 作者简介：
- 基金信息：
  
  the Project of Hubei Provincial Health Commission(WJ2019Q057)
- DOI：10.16016/j.1000-5404.202106071
  CLC： R341;R363.21;R542.23
- Received：08 June 2021，
  
  Revised：2021-09-20，
  
  Published：30 December 2021
- 稿件说明：
移动端阅览
Qian ZHAO, Yong ZOU, Bo LIU, et al. Role of runt-related transcription factor 3 in myocardial fibrosis in mice[J]. Journal of Third Military Medical University, 2021, 43(24): 2640-2647.
DOI：

Qian ZHAO, Yong ZOU, Bo LIU, et al. Role of runt-related transcription factor 3 in myocardial fibrosis in mice[J]. Journal of Third Military Medical University, 2021, 43(24): 2640-2647. DOI： 10.16016/j.1000-5404.202106071.

摘要

目的

探讨Runt相关转录因子3(runt related transcription factor 3，Runx3)在心肌梗死(myocardial infarction，MI)后心脏成纤维细胞分化、Ⅰ型胶原(collagen Ⅰ

ColⅠ)合成和增殖中的作用。

方法

选择C57BL/6J雄性小鼠为研究对象。分为模型组(MI)和假手术组(Sham)，MI组采用结扎左冠状动脉建立MI模型，分别在建模后7、14、28 d处死小鼠；Sham组不结扎左冠状动脉。另将Sham组和MI组小鼠建模前尾静脉分别注射Runx3小干扰RNA(si-Runx3)或阴性对照siRNA(si-NC)，建模后28 d处死，分为Sham+si-NC组、Sham+si-Runx3组、MI 28 d+si-NC组和MI 28 d+si-Runx3组，每组8只。应用免疫荧光染色和Western blot检测小鼠心脏中Runx3、α-SMA和ColⅠ的水平，采用超声心动图评价心功能，免疫组化方法检测Ki67阳性细胞。体外实验中，从1~3 d龄的C57BL/6小鼠中分离心脏成纤维细胞，观察TGF-β1以不同浓度和刺激时间对细胞中Runx3蛋白影响，并分析Runx3基因敲除对心脏成纤维细胞的ColⅠ合成和增殖的响。

结果

免疫荧光染色和Western blot结果，随着MI时间的延长，小鼠纤维化区域Runx3、α-SMA和ColⅠ表达上调。MI后28 d，与注射si-NC小鼠相比，注射si-Runx3小鼠左心室射血分数(ejection fraction，EF)、缩短分数(shortened fraction，FS)、心输出量(cardiac output，CO)。上调(

0.05)，而舒张末期左心室内径(decreased left ventricular end-diastolic diameter，LVIDd)和收缩期左心室内径(decreased left ventricular，LVIDs)下调(

0.05)。MI后28 d，MI诱导的Runx3和α-SMA表达和对Ki67阳性细胞增殖作用被si-Runx3阻断。TGF-β1以浓度和时间依赖性方式上调Runx3蛋白表达；Runx3基因敲除不仅抑制TGF-β1诱导的心脏成纤维细胞分化，减少ColⅠ合成，而且减弱了心

脏成纤维细胞增殖。

结论

Runx3参与了心脏成纤维细胞分化和增殖。

Abstract

Objective

To investigate the role of runt related transcription factor 3 (Runx3) in cardiac fibroblasts differentiation

proliferation as well as synthesis of type I collagen after myocardial infarction (MI) in mice.

Methods

Male C57BL/6J mice were selected and randomly divided into model group (MI group

treated with ligation of left coronary artery) and Sham group

and were sacrificed at the 7th

14th and 28th days after modeling (

=6 for each group)

respectively. In addition

Runx3 small interfering RNA (si-Runx3) was injected

via

the tail vein of mice before modeling to achieve Runx3 knockdown

negative siRNA (si-NC) as the matched control. According to the treatment they received

the mice were subsequently allocated into Sham+si-NC group

Sham+si-Runx3 group

MI 28 d+si-NC group and MI 28 d+si-Runx3 group (

=8)

and were sacrificed in 28 d after modeling. The levels of Runx3

alpha-smooth muscle actin (α-SMA) and type I collagen in the heart were determined by immunofluorescence staining and Western blotting. The cardiac function was evaluated by echocardiography

and Ki67 positive cells were observed using immunohistochemical assay. After cardiac fibroblasts were isolated from C57BL/6 mice and cultured for 1~3 d

and the effects of TGF-β1 at different concentrations and for different stimulation times on Runx3 protein were observed in the cells. The effects of Runx3 gene knockout on type I collagen synthesis and proliferation of cardiac fibroblasts were also analyzed.

Results

Immunofluorescence staining and Western blotting showed that the expression levels of Runx3

α-SMA and type I collagen in the fibrosis regions were up-regulated with the extended duration of MI. As compared with the MI 28 d+si-NC group

the MI 28

d+si-Runx3 group had increased left ventricular ejection fraction (LVEF)

fractional shortening (FS) and cardiac output (

0.05)

and decreased left ventricular end-diastolic diameter (LVIDd) and decreased left ventricular (LVIDs) (

0.05) in 28 d after MI; the expression levels of Runx3 and α-SMA and the proliferation of Ki67 positive cells induced by MI were blocked by si-Runx3. TGF-β1 up-regulated Runx3 expression in a concentration- and time-dependent manner. Runx3 knockdown not only inhibited TGF-β1-induced differentiation of cardiac fibroblasts

reduced type I collagen synthesis

but also attenuated cardiac fibroblasts proliferation.

Conclusions

Runx3 is involved in the cardiac fibroblast differentiation and proliferation.

关键词

Keywords

references

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