Runt相关转录因子3在小鼠心肌纤维化中的作用

赵谦; 邹勇; 刘波; 余信强; 张苏川

doi:10.16016/j.1000-5404.202106071

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Runt相关转录因子3在小鼠心肌纤维化中的作用

基础医学 | 更新时间：2026-01-09

- Runt相关转录因子3在小鼠心肌纤维化中的作用
- Role of runt-related transcription factor 3 in myocardial fibrosis in mice
- 第三军医大学学报 2021年43卷第24期页码：2640-2647
- 作者机构：
  
  430015 武汉，江汉大学附属医院心血管内科
- 作者简介：
  
  [ 赵谦，E-mail：qianazhao578@163.com ZHAO Qian, E-mail: qianazhao578@163.com
- 基金信息：
  
  湖北省卫生健康委员会课题(WJ2019Q057)
- DOI：10.16016/j.1000-5404.202106071
  中图分类号： R341;R363.21;R542.23
- 收稿：2021-06-08，
  
  修回：2021-09-20，
  
  纸质出版：2021-12-30
- 稿件说明：
移动端阅览
赵谦, 邹勇, 刘波, 等. Runt相关转录因子3在小鼠心肌纤维化中的作用[J]. 第三军医大学学报, 2021,43(24):2640-2647.

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.
赵谦, 邹勇, 刘波, 等. Runt相关转录因子3在小鼠心肌纤维化中的作用[J]. 第三军医大学学报, 2021,43(24):2640-2647. DOI： 10.16016/j.1000-5404.202106071.

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

陶逸菁, 夏智丽, 高程洁, 等. 急性心肌梗死再血管化成功后预测左心室重构相关生物标志物的分析[J]. 上海交通大学学报(医学版), 2019, 39(1): 60-64. DOI: 10.3969/Jissn.1674-8115.2019.01.011.

TAO Y J, XIA Z L, GAO C J, et al . Analysis of biomarkers for predicting left ventricular remodeling after successful revascularization of acute myocardial infarction[J ] . J Shanghai Jiaotong Univ (Med Sci), 2019, 39(1): 60-64. DOI: 10.3969/Jissn.1674-8115.2019.01.011.

孙润民, 王琼英, 齐苗苗, 等. 急性心肌梗死后心力衰竭预测因素的研究进展[J]. 中华心血管病杂志, 2019, 47(11): 923-926. DOI: 10.3760/cma.j.issn.0253-3758.2019.11.015.

SUN R M, WANG Q Y, QI M M, et al . Research progress on the prediction of heart failure post acute myocardial infarction: clinical feature, risk score and biomarkers[J ] . Chin J Cardiol, 2019, 47(11): 923-926. DOI: 10.3760/cma.j.issn.0253-3758.2019.11.015.

吴学平, 李志宏, 王新艳, 等. 心肌梗死后心脏修复与心肌细胞再生的研究进展[J]. 中国动脉硬化杂志, 2019, 27(10): 899-904.

WU X P, LI Z H, WANG X Y, et al . Recent advances in cardiac repair and regeneration after myocardial infarction[J ] . Chin J Arterioscler, 2019, 27(10): 899-904.

郑海清, 应苗法, 顾胜龙, 等. 心肌纤维化的信号转导机制及新型抑制剂的研究进展[J]. 中国细胞生物学学报, 2019, 41(2): 268-274.

ZHENG H Q, YING M F, GU S L, et al . Molecular signaling mechanism behind myocardial fibrosis and advances in novel inhibitors[J ] . Chin J Cell Biol, 2019, 41(2): 268-274.

LEE S H, HYEON D Y, YOON S H, et al . RUNX3 methylation drives hypoxia-induced cell proliferation and antiapoptosis in early tumorigenesis[J ] . Cell Death Differ, 2021, 28(4): 1251-1269. DOI: 10.1038/s41418-020-00647-1.

KIM B R, KANG M H, KIM J L, et al . RUNX3 inhibits the metastasis and angiogenesis of colorectal cancer[J ] . Oncol Rep, 2016, 36(5): 2601-2608. DOI: 10.3892/or.2016.5086.

ZHENG K, YU J, CHEN Z, et al . Ethanol promotes alcohol-related colorectal cancer metastasis via the TGF-β/RUNX3/Snail axis by inducing TGF-β1 upregulation and RUNX3 cytoplasmic mislocalizatio n[J ] . EBioMedicine, 2019, 50: 224-237. DOI: 10.1016/j.ebiom.2019.11.011.

LIU Y H, ZOU J, LI B G, et al . RUNX3 modulates hypoxia-induced endothelial-to-mesenchymal transition of human cardiac microvascular endothelial cells[J ] . Int J Mol Med, 2017, 40(1): 65-74. DOI: 10.3892/ijmm.2017.2998.

TAO L, BEI Y, CHEN P, et al . Crucial role of miR-433 in regulating cardiac fibrosis[J ] . Theranostics, 2016, 6(12): 2068-2083. DOI: 10.7150/thno.15007.

陈羽斐, 沈伟, 施海明. 急性心肌梗死逆向加速动脉硬化进程的机制研究进展[J]. 中华医学杂志, 2019, 99(1): 72-74. DOI: 10.3760/cma.j.issn.0376-2491.2019.01.017.

CHEN Y F, SHEN W, SHI H M. Research progress on Mechanism of reverse acceleration of atherosclerosis in acute myocardial infarction[J]. Natl Med J China, 2019, 99(1): 72-74. DOI: 10.3760/cma.j.issn.0376-2491.2019.01.017.

蓝庆肃, 韩冰, 彭瑜, 等. 骨膜蛋白与梗死后心肌纤维化的相关性研究进展[J]. 临床心血管病杂志, 2020, 36(12): 1154-1158. DOI: 10.13201/j.issn.1001-1439.2020.12.018.

LAN Q S, HAN B, PENG Y, et al . Advanced progress of the relationship between periostin and post-infraction myocardial fibrosis[J ] . J Clin Cardiol, 2020, 36(12): 1154-1158. DOI: 10.13201/j.issn.1001-1439.2020.12.018.

TALMAN V, RUSKOAHO H. Cardiac fibrosis in myocardial infarction-from repair and remodeling toregeneration[J]. Cell Tissue Res, 2016, 365(3): 563-581. DOI: 10.1007/s00441-016-2431-9.

XIAO Z, TIAN Y, JIA Y, et al . RUNX3 inhibits the invasion and migration of esophageal squamous cell carcinoma by reversing the epithelial-mesenchymal transition through TGF-β/Smad signaling[J ] . Oncol Rep, 2020, 43(4): 1289-1299. DOI: 10.3892/or.2020.7508.

左银龙, 周易, 黄珍谷, 等. runx3对人骨肉瘤143B细胞增殖、迁移和侵袭的影响及机制[J]. 南昌大学学报(医学版), 2020, 60(5): 22-27, 33. DOI: 10.13764/j.cnki.ncdm.2020.05.005.

ZUO Y L, ZHOU Y, HUANG Z G, et al . Effects of Runx3 on proliferation, migration and invasion and its possible mechanism in human 143B osteosarcoma cells[J ] . J Nanchang Univ (Med Sci), 2020, 60(5): 22-27, 33. DOI: 10.13764/j.cnki.ncdm.2020.05.005.

BAI X D, HAN G H, LIU Y, et al . MiRNA-20a-5p promotes the growth of triple-negative breast cancer cells through targeting RUNX3[J ] . Biomed Pharmacother, 2018, 103: 1482-1489. DOI: 10.1016/j.biopha.2018.04.165.

ZHANG Y, WANG S, LAI Q, et al . Cancer-associated fibroblasts-derived exosomal miR-17-5p promotes colorectal cancer aggressive phenotype by initiating a RUNX3/MYC/TGF-β1 positive feedback loop[J ] . Cancer Lett, 2020, 491: 22-35. DOI: 10.1016/j.canlet.2020.07.023.

SHI L, LIU B Y, WANG X, et al . RUNX3-dependent oxidative epithelial-to-mesenchymal transition in methamphetamine-induced chronic lung injury[J ] . Cell Stress Chaperones, 2020, 25(5): 793-802. DOI: 10.1007/s12192-020-01133-w.

HUANG S, CHEN B, SU Y, et al . Distinct roles of myofibroblast-specific Smad2 and Smad3 signaling in repair and remodeling of the infarcted heart[J ] . J Mol Cell Cardiol, 2019, 132: 84-97. DOI: 10.1016/j.yjmcc.2019.05.006.

WANG L, JIANG P, HE Y, et al . A novel mechanism of Smads/miR-675/TGFβR1 axis modulating the proliferation and remodeling of mouse cardiac fibroblasts[J ] . J Cell Physiol, 2019, 234(11): 20275-20285. DOI: 10.1002/jcp.28628.

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