The role of cathepsin K in cardiac dysfunction in mice with alcoholic cardiomyopathy and its damage mechanism

Yifeng TANG; Zhijiang LIU; Guanxue XU; Ranzun ZHAO; Wenming CHEN; Bei SHI

doi:10.16016/j.2097-0927.202112025

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The role of cathepsin K in cardiac dysfunction in mice with alcoholic cardiomyopathy and its damage mechanism

Basic Medicine | 更新时间：2026-01-09

- The role of cathepsin K in cardiac dysfunction in mice with alcoholic cardiomyopathy and its damage mechanism
- Journal of Army Medical University Vol. 44, Issue 13, Pages: 1330-1337(2022)
- 作者机构：
  
  1.553001 贵州六盘水，六盘水市人民医院心血管内科
  2.563000 贵州遵义，遵义医科大学附属医院心血管内科
- 作者简介：
- 基金信息：
  
  the Project of Health Scientific Research Foundation of Guizhou Province(20190124)
- DOI：10.16016/j.2097-0927.202112025
  CLC： R345;R363.21;R542.2
- Received：03 December 2021，
  
  Revised：2022-02-28，
  
  Published：15 July 2022
- 稿件说明：
移动端阅览
Yifeng TANG, Zhijiang LIU, Guanxue XU, et al. The role of cathepsin K in cardiac dysfunction in mice with alcoholic cardiomyopathy and its damage mechanism[J]. Journal of Army Medical University, 2022, 44(13): 1330-1337.
DOI：

Yifeng TANG, Zhijiang LIU, Guanxue XU, et al. The role of cathepsin K in cardiac dysfunction in mice with alcoholic cardiomyopathy and its damage mechanism[J]. Journal of Army Medical University, 2022, 44(13): 1330-1337. DOI： 10.16016/j.2097-0927.202112025.

摘要

目的

探讨组织蛋白酶K(cathepsin K，CatK)在酒精性心肌病(alcoholic cardiomyopathy，ACM)小鼠心功能不全中的作用及其损伤机制。

方法

48只8周龄C57BL/6J雄性小鼠按随机数字表法分为对照组(

=12，等热量麦芽糖糊精对照液体饲料喂养)和模型组(

=36，4%乙醇Lieber-DeCarli液体饲料喂养)。8周后，以超声心动图心脏收缩功能下降的模型组小鼠为ACM，按随机数字表法分为: ACM组、sh-NC组、sh-CatK组，每组12只。分别向sh-NC组和sh-CatK组小鼠鼠尾静脉注射1×10

vg(100 μL)Scramble-shRNA和CatK-shRNA的重组腺病毒。在第12周末，采用M型超

声心动图测量心功能。免疫组织化学检测CatK和Nox4蛋白表达。体外实验以H9c2心肌细胞为研究对象，评价CatK-shRNA介导的CatK沉默对乙醇诱导的心肌细胞损伤的影响。采用蛋白质印迹法分析AMPK/Nox4信号通路蛋白表达；采用流式细胞仪分析H9c2细胞凋亡情况。应用荧光染料二氢乙胺测定小鼠心肌组织和体外培养的H9c2细胞的活性氧水平。

结果

ACM组心肌细胞表面积，心肌纤维化面积，心肌组织中CatK、Nox4蛋白表达和超氧化物的产生均较对照组显著增加(

0.05)。与ACM组相比，sh-CatK组心肌细胞表面积，心肌纤维化面积，心肌组织中CatK、Nox4蛋白表达和超氧化物的产生均显著降低(

0.05)。体外结果显示，随着暴露于乙醇中的时间不断延长，H9c2细胞中CatK蛋白表达不断增加(

0.05)。CatK抑制处理显著降低了H9c2细胞凋亡和氧化应激水平(

0.05)，并上调了AMPK磷酸化(

0.05)，进而导致Nox4蛋白、纤维细胞胶原COL-1和TGF-β表达降低(

0.05)。

结论

CatK参与ACM小鼠心功能不全的病理过程，其机制与促进AMPK/Nox4信号通路介导的氧化应激有关。

Abstract

Objective

To investigate the role of cathepsin K (CatK) in cardiac dysfunction in alcoholic cardiomyopathy (ACM) mice and its damage mechanism.

Methods

Forty-eight C57BL/6J male mice (8 weeks old) were randomly divided into control group (

=12

fed with isocaloric maltodextrin control liquid diet) and model group (

=36

fed with 4% ethanol Lieber-DeCarli liquid diet). After 8 weeks

the mice in the model group identified with decreased cardiac systolic function by echocardiography were regarded as ACM mice

and they were randomly divided into ACM group

sh-NC group

and sh-CatK group

with 12 mice in each group. The mice in the sh-NC group and the sh-CatK group were injected with 1×10

vg (100 μL) of recombinant adenoviruses containing scramble-shRNA or CatK-shRNA

via

tail vein

respectively. At the end of the 12th week

cardiac function was measured by M-mode echocardiography. The expression of CatK and Nox4 was detected by immunohistochemical assay. In

in vitro

study

H9c2 cardiomyocytes were used to evaluate the effect of CatK silencing on alcohol-induced cardiomyocyte injury. Western blotting w

as used to analyze the protein expression of AMPK/Nox4 signaling pathway. Flow cytometry was employed to analyze the apoptosis of H9c2 cells. The levels of reactive oxygen species in mouse myocardium and H9c2 cells cultured were measured using fluorescent dye dihydroethidium.

Results

The surface area of myocardial cells

area of myocardial fibrosis

protein levels of CatK and Nox4 and production of superoxide were significantly increased in the ACM group compared with the control group (

0.05)

and those indicators in the sh-CatK group were significantly decreased than the ACM group (

0.05). Sh-CatK transfection significantly reversed the above changes in ACM mice (

0.05).

In vitro

results showed that the expression of CatK protein in H9c2 cells was increased with the prolonged exposure to ethanol (

0.05). CatK inhibition significantly reduced the apoptostic rate and oxidative stress levels (

0.05)

and up-regulated AMPK phosphorylation in H9c2 cells (

0.05)

resulting in decreased expression of Nox4 protein

collagen COL-1 and TGF-β (

0.05).

Conclusion

CatK is involved in the pathological process of cardiac dysfunction in the ACM mice

and its mechanism might be related to the promotion of oxidative stress mediated by the AMPK/Nox4 signaling pathway.

关键词

Keywords

references

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