Ubiquitin C-terminal hydrolase 1 inhibits the ubiquitination of CaMK Ⅱ and promots the occurrence of atrial fibrillation in rats

Liying LI; Li FANG; Heng QI; Bin ZOU; Cong YUAN; Yuxuan GUO

doi:10.16016/j.2097-0927.202204211

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Ubiquitin C-terminal hydrolase 1 inhibits the ubiquitination of CaMK Ⅱ and promots the occurrence of atrial fibrillation in rats

Basic Medicine | 更新时间：2026-02-13

- Ubiquitin C-terminal hydrolase 1 inhibits the ubiquitination of CaMK Ⅱ and promots the occurrence of atrial fibrillation in rats
- Journal of Army Medical University Vol. 44, Issue 22, Pages: 2275-2284(2022)
- 作者机构：
  
  410000 长沙，长沙市第一医院心血管内科二病区
- 作者简介：
- 基金信息：
  
  the Natural Science Foundation of Hunan Province of China(2018JJ6127)
- DOI：10.16016/j.2097-0927.202204211
  CLC： R345;R363.21;R541.75
- Received：29 April 2022，
  
  Revised：2022-08-10，
  
  Published：30 November 2022
- 稿件说明：
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Liying LI, Li FANG, Heng QI, et al. Ubiquitin C-terminal hydrolase 1 inhibits the ubiquitination of CaMK Ⅱ and promots the occurrence of atrial fibrillation in rats[J]. Journal of Army Medical University, 2022, 44(22): 2275-2284.
DOI：

Liying LI, Li FANG, Heng QI, et al. Ubiquitin C-terminal hydrolase 1 inhibits the ubiquitination of CaMK Ⅱ and promots the occurrence of atrial fibrillation in rats[J]. Journal of Army Medical University, 2022, 44(22): 2275-2284. DOI： 10.16016/j.2097-0927.202204211.

摘要

目的

分析泛素C末端水解酶1(ubiquitin C-terminal hydrolase 1，UCHL1)在促进心房颤动(atrial fibrillation，AF)发生中的作用和潜在分子机制。

方法

40只雄性SD大鼠(8周龄，体质量240~260 g)采用随机数字表法分为4组，每组10只：Sham+DMSO组、Sham+LDN(UCHL1抑制剂)组、AF+DMSO组和AF+LDN组。分组后，AF+DMSO组和AF+LDN组通过皮下植入血管紧张素Ⅱ(angiotensinⅡ，AngⅡ)渗透性微型泵诱导AF模型。4周后，使用心电图记录监测AF的持续时间；Masson三色染色左心房组织以观察纤维化水平；采用超声心动图检查心脏功能；从1~3日龄的SD大鼠心脏中分离新生大鼠心肌细胞(neonatal rat cardiomyocytes，NRCM)，通过在NRCM细胞中过表达或敲低UCHL1验证其作用；进行免疫沉淀质谱分析，评估UCHL1对NRCM中钙/钙调素依赖性蛋白激酶Ⅱ(calcium/calmodulin-dependent protein kinaseⅡ，CaMKⅡ)的功能调节机制；采用蛋白质印迹分析纤维化蛋白表达。

结果

在AF组大鼠心脏组织和AngⅡ处理的NRCM中，UCHL1蛋白表达增加。与AF+DMSO组相比，AF+LDN组大鼠的心脏射血分数(ejection fraction，EF)和缩短分数(fractional shortening，FS)显著升高(

0.01)，

Masson三色染色纤维化程度、促纤维化蛋白α-SMA、ColⅠ表达和AF的持续时间显著降低(

0.01)。体外实验中，UCHL1的敲低可减轻AngⅡ诱导的NRCM纤维化(

0.05)。免疫沉淀质谱显示UCHL1与AngⅡ刺激的NRCM中的CaMKⅡ相互作用，并通过去泛素化促进其上调。CaMKⅡ过表达降低了UCHL1敲低对AngⅡ刺激的NRCM中纤维化反应的抑制作用(

0.05)。

结论

UCHL1与CaMK Ⅱ相互作用，并通过去泛素化上调CaMK Ⅱ，从而促进AF的心脏纤维化。

Abstract

Objective

To analyze the role and molecular mechanisms underlying of ubiquitin C-terminal hydrolase 1 (UCHL1) in promoting atrial fibrillation (AF).

Methods

Forty 8-week-old male SD rats (240~260 g) were randomly divided into four groups

with 10 rats in each group: Sham+DMSO group

Sham+ LDN (the inhibitor of UCHL1) group

AF+DMSO group and AF+LDN group. AF model was induced by subcutaneously implanted osmotic micropump of angiotensin Ⅱ (AngⅡ) in both AF+DMSO group and AF+LDN group. After 4 weeks

the duration of AF was monitored and recorded by electrocardiography. The level of fibrosis was observed by Masson-trichrome staining of left atrial tissues. Heart function was evaluated by ultrasonic cardiography. Neonatal rat cardiomyocytes (NRCM) were isolated from the hearts of SD rats aged 1~3 d of which effect was verified by overexpression or knockdown of UCHL1 in NRCM cells. Immunoprecipitation mass spectrometry was used to evaluate the functional regulatory mechanism of UCHL1 on calcium/calmodulin-dependent protein kinaseⅡ (CaMKⅡ) in NRCM. The expression of fibrotic protein was analyzed by Western blot assay.

Results

The expression of UCHL1 protein was increased in cardiac tissues of AF rats and in AngⅡ-treated NRCM. Compared with the AF+DMSO group

the cardiac ejection fraction (EF) and fractional shortening (FS) of the AF+LDN group were significantly increased (

0.01)

while the degree of fibrosis

expression of α-SMA and ColⅠ and duration of AF were significantly decreased(

0.01).

In vitro

inhibition and knockdown of UCHL1 attenuated AngⅡ-induced fibrosis of NRCM (

0.05). Immunoprecipitation mass spectrometry revealed that UCHL1 interacted with CaMKⅡ in NRCM simulated by AngⅡand promoted the upregulation of UCHL1 through deubiquitination. CaMKⅡ overexpression reduced the inhibitory effect of UCHL1 knockdown on the fibrotic response in AngⅡ-induced NRCM (

0.05).

Conclusion

UCHL1 interacts with CaMKⅡ and upregulates CaMKⅡ through deubiquitination

thereby promoting cardiac fibrosis in AF.

关键词

Keywords

references

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