浏览全部资源
扫码关注微信
泛素C末端水解酶1抑制CaMKⅡ泛素化促进心房颤动
Ubiquitin C-terminal hydrolase 1 inhibits the ubiquitination of CaMK Ⅱ and promots the occurrence of atrial fibrillation in rats
- 陆军军医大学学报 2022年44卷第22期 页码：2275-2284
- 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：
[方立，E-mail: skyer200207@163.com FANG Li, E-mail: skyer200207@163.com
- 基金信息：湖南省自然科学基金(2018JJ6127)
DOI：10.16016/j.2097-0927.202204211
中图分类号： R345;R363.21;R541.75收稿：2022-04-29，
修回：2022-08-10，
纸质出版：2022-11-30
- 稿件说明：
移动端阅览
李莉英, 方立, 祁珩, 等. 泛素C末端水解酶1抑制CaMKⅡ泛素化促进心房颤动[J]. 陆军军医大学学报, 2022,44(22):2275-2284.
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.
李莉英, 方立, 祁珩, 等. 泛素C末端水解酶1抑制CaMKⅡ泛素化促进心房颤动[J]. 陆军军医大学学报, 2022,44(22):2275-2284. DOI： 10.16016/j.2097-0927.202204211.
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
2
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
2
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
2
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(
P
<
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(
P
<
0.01)。体外实验中，UCHL1的敲低可减轻AngⅡ诱导的NRCM纤维化(
P
<
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(
P
<
0.05)。
结论
2
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
2
To analyze the role and molecular mechanisms underlying of ubiquitin C-terminal hydrolase 1 (UCHL1) in promoting atrial fibrillation (AF).
Methods
2
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
2
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 (
P
<
0.01)
while the degree of fibrosis
expression of α-SMA and ColⅠ and duration of AF were significantly decreased(
P
<
0.01).
In vitro
inhibition and knockdown of UCHL1 attenuated AngⅡ-induced fibrosis of NRCM (
P
<
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 (
P
<
0.05).
Conclusion
2
UCHL1 interacts with CaMKⅡ and upregulates CaMKⅡ through deubiquitination
thereby promoting cardiac fibrosis in AF.
关键词
Keywords
references
李泰然, 陈桂英. 心外膜脂肪组织与心房颤动的分子机制研究进展[J]. 生物化学与生物物理进展, 2021, 48(9): 1016-1022. DOI: 10.16476/j.pibb.2020.0428.
LI T R, CHEN G Y. Research progress on molecular mechanism of epicardial adipose tissue and atrial fibrillation[J]. Prog Biochem Biophys, 2021, 48(9): 1016-1022. DOI: 10.16476/j.pibb.2020.0428.
JANUARY C T, WANN L S, CALKINS H, et al . 2019 AHA/ACC/HRS focused update of the 2014 AHA/ACC/HRS guideline for the management of patients with atrial fibrillation: a report of the American college of cardiology/American heart association task force on clinical practice guidelines and the heart rhythm society[J ] . J Am Coll Cardiol, 2019, 74(1): 104-132. DOI: 10.1016/j.jacc.2019.01.011.
YUAN Y, ZHAO J, GONG Y T, et al . Autophagy exacerbates electrical remodeling in atrial fibrillation by ubiquitin-dependent degradation of L-type calcium channel[J ] . Cell Death Dis, 2018, 9(9): 873. DOI: 10.1038/s41419-018-0860-y.
刘志浩, 倪晶宇, 郭蕊, 等. 翻译后修饰对SERCA2a的作用在心血管疾病中的研究进展[J]. 中华心血管病杂志, 2020, 48(2): 164-167. DOI: 10.3760/cma.j.issn.0253-3758.2020.02.015.
LIU Z H, NI J Y, GUO R, et al . Advances in research on the effects of post-translational modification on SERCA2a in cardiovascular diseases[J ] . Chin J Cardiol, 2020, 48(2): 164-167. DOI: 10.3760/cma.j.issn.0253-3758.2020.02.015.
AHN H J, JUNG S, YOU Y, et al . The value of cerebrospinal fluid ubiquitin C-terminal hydrolase-L1 protein as a prognostic predictor of neurologic outcome in post-cardiac arrest patients treated with targeted temperature management[J ] . Resuscitation, 2020, 151: 50-58. DOI: 10.1016/j.resuscitation.2020.03.022.
MAO R D, TAN X, XIAO Y, et al . Ubiquitin C-terminalhydrolase L1 promotes expression of programmed cell death-ligand 1 in non-small-cell lung cancer cells[J ] . Cancer Sci, 2020, 111(9): 3174-3183. DOI: 10.1111/cas.14529.
GONG Z, YE Q, WU J W, et al . UCHL1 inhibition attenuates cardiac fibrosis via modulation of nuclear factor-κB signaling in fibroblasts[J ] . Eur J Pharmacol, 2021, 900: 174045. DOI: 10.1016/j.ejphar.2021.174045..
XIAO Z Z, REDDY D P K, XUE C Q, et al . Profiling of miR-205/P4HA3 following angiotensin Ⅱ-induced atrial fibrosis: implications for atrial fibrillation[J ] . Front Cardiovasc Med, 2021, 8: 609300. DOI: 10.3389/fcvm.2021.609300.
USUI S, CHIKATA A, TAKATORI O, et al . Endogenous muscle atrophy F-box is involved in the development of cardiac rupture after myocardial infarction[J ] . J Mol Cell Cardiol, 2019, 126: 1-12. DOI: 10.1016/j.yjmcc.2018.11.002.
MCMACKEN G, LOCHMVLLER H, BANSAGI B, et al . Behr syndrome and hypertrophic cardiomyopathy in a family with a novel UCHL1 deletion[J ] . J Neurol, 2020, 267(12): 3643-3649. DOI: 10.1007/s00415-020-10059-3.
LU L, CAO L, LIU Y H, et al . Angiotensin (ang) 1-7 inhibits ang Ⅱ-induced atrial fibrosis through regulating the interaction of proto-oncogene tyrosine-protein kinase Src (c-Src) and Src homology region 2 domain-containing phosphatase-1 (SHP-1)[J ] . Bioengineered, 2021, 12(2): 10823-10836. DOI: 10.1080/21655979.2021.1967035.
ZHAN Y G, ABE I, NAKAGAWA M, et al . A traditional herbal medicine rikkunshito prevents angiotensin Ⅱ-induced atrial fibrosis and fibrillation[J ] . J Cardiol, 2020, 76(6): 626-635. DOI: 10.1016/j.jjcc.2020.07.001.
WANG Q, YU Y, ZHANG P P, et al . The crucial role of activin A/ALK4 pathway in the pathogenesis of AngⅡ-induced atrial fibrosis and vulnerability to atrial fibrillation[J ] . Basic Res Cardiol, 2017, 112(4): 47. DOI: 10.1007/s00395-017-0634-1.
LI X B, HATTORI A, TAKAHASHI S, et al . Ubiquitin carboxyl-terminal hydrolase L1 promotes hypoxia-inducible factor 1-dependent tumor cell malignancy in spheroid models[J ] . Cancer Sci, 2020, 111(1): 239-252. DOI: 10.1111/cas.14236.
DOWNEY A, OLCOTT M, SPECTOR D, et al . Stable knockout of lanthionine synthase C-like protein-1 (LanCL1) from HeLa cells indicates a role for LanCL1 in redox regulation of deubiquitinating enzymes[J ] . Free Radic Biol Med, 2020, 161: 115-124. DOI: 10.1016/j.freeradbiomed.2020.10.006.
GAO H B, HARTNETT S, LI Y F. Ubiquitin C-terminal hydrolase L1 regulates myoblast proliferation and differentiation[J]. Biochem Biophys Res Commun, 2017, 492(1): 96-102. DOI: 10.1016/j.bbrc.2017.08.027.
WADE B E, ZHAO J R, MA J, et al . Hypoxia-induced alterations in the lung ubiquitin proteasome system during pulmonary hypertension pathogenesis[J ] . Pulm Circ, 2018, 8(3): 2045894018788267. DOI: 10.1177/2045894018788267.
COSTES S, BERTRAND G, RAVIER M A. Mechanisms of beta-cell apoptosis in type 2 diabetes-prone situations and potential protection by GLP-1-based therapies[J]. Int J Mol Sci, 2021, 22(10): 5303. DOI: 10.3390/ijms22105303.
李健兰, 陈灿. 心房颤动患者心房纤维化检测方式的研究进展[J]. 山东医药, 2020, 60(13): 85-88. DOI: 10.3969/j.issn.1002-266X.2020.13.025.
LI J L, CHEN C. Research progress on the detection method of atrial fibrosis in patients with atrial fibrillation[J]. Shandong Med J, 2020, 60(13): 85-88. DOI: 10.3969/j.issn.1002-266X.2020.13.025.
马婧, 刘晓莉, 乔德才. 钙/钙调蛋白依赖性蛋白激酶Ⅱ在突触可塑性和神经精神疾病中的作用[J]. 中国药理学通报, 2018, 34(11): 1485-1488. DOI: 10.3969/j.issn.1001-1978.2018.11.002.
MA J, LIU X L, QIAO D C. Role of calcium/calmodulin-dependent protein kinase Ⅱ in synaptic plasticity and neuropsychiatric disorders[J]. Chin Pharmacol Bull, 2018, 34(11): 1485-1488. DOI: 10.3969/j.issn.1001-1978.2018.11.002.
LIU T, XIONG F, QI X Y, et al . Altered calcium handling produces reentry-promoting action potential alternans in atrial fibrillation-remodeled hearts[J ] . JCI Insight, 2020, 5(8): e133754. DOI: 10.1172/jci.insight.133754.
GRANDI E, DOBREV D. Non-ion channel therapeutics for heart failure and atrial fibrillation: are CaMKⅡ inhibitors ready for clinical use?[J]. J Mol Cell Cardiol, 2018, 121: 300-303. DOI: 10.1016/j.yjmcc.2017.10.010.
0
浏览量
0
下载量
0
CSCD
- 网络PDF下载
- Meta-XML下载
- BibTeX下载
- Endnote下载
- RefMan 下载
- NoteExpress下载
- NoteFirst下载
关联资源
相关文章
相关作者
相关机构
- 技术支持由北京北大方正电子有限公司提供 京ICP备09064830号-19
京公网安备11010802024621 - 本系统建议在Chrome、 IE9+ 以上版本浏览器阅读本站内容，360浏览器请切换至极速模式
- Cookies帮助我们提供服务并提供个性化体验。使用本网站，即表示您同意我们使用Cookies
