浏览全部资源
扫码关注微信
组织蛋白酶K在酒精性心肌病小鼠心功能不全中的作用及其损伤机制研究
The role of cathepsin K in cardiac dysfunction in mice with alcoholic cardiomyopathy and its damage mechanism
- 陆军军医大学学报 2022年44卷第13期 页码：1330-1337
- 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：
[石蓓，E-mail：wzjlfl2604@163.com SHI Bei, E-mail: wzjlfl2604@163.com
- 基金信息：贵州省卫生健康科研基金项目(20190124)
DOI：10.16016/j.2097-0927.202112025
中图分类号： R345;R363.21;R542.2收稿：2021-12-03，
修回：2022-02-28，
纸质出版：2022-07-15
- 稿件说明：
移动端阅览
唐一锋, 刘志江, 许官学, 等. 组织蛋白酶K在酒精性心肌病小鼠心功能不全中的作用及其损伤机制研究[J]. 陆军军医大学学报, 2022,44(13):1330-1337.
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.
唐一锋, 刘志江, 许官学, 等. 组织蛋白酶K在酒精性心肌病小鼠心功能不全中的作用及其损伤机制研究[J]. 陆军军医大学学报, 2022,44(13):1330-1337. DOI： 10.16016/j.2097-0927.202112025.
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
2
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
2
48只8周龄C57BL/6J雄性小鼠按随机数字表法分为对照组(
n
=12，等热量麦芽糖糊精对照液体饲料喂养)和模型组(
n
=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
9
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
2
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(
P
<
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(
P
<
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(
P
<
0.05)。CatK抑制处理显著降低了H9c2细胞凋亡和氧化应激水平(
P
<
0.05)，并上调了AMPK磷酸化(
P
<
0.05)，进而导致Nox4蛋白、纤维细胞胶原COL-1和TGF-β表达降低(
P
<
0.05)。
结论
2
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
2
To investigate the role of cathepsin K (CatK) in cardiac dysfunction in alcoholic cardiomyopathy (ACM) mice and its damage mechanism.
Methods
2
Forty-eight C57BL/6J male mice (8 weeks old) were randomly divided into control group (
n
=12
fed with isocaloric maltodextrin control liquid diet) and model group (
n
=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
9
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
2
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 (
P
<
0.05)
and those indicators in the sh-CatK group were significantly decreased than the ACM group (
P
<
0.05). Sh-CatK transfection significantly reversed the above changes in ACM mice (
P
<
0.05).
In vitro
results showed that the expression of CatK protein in H9c2 cells was increased with the prolonged exposure to ethanol (
P
<
0.05). CatK inhibition significantly reduced the apoptostic rate and oxidative stress levels (
P
<
0.05)
and up-regulated AMPK phosphorylation in H9c2 cells (
P
<
0.05)
resulting in decreased expression of Nox4 protein
collagen COL-1 and TGF-β (
P
<
0.05).
Conclusion
2
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
WANG S Y, REN J. Role of autophagy and regulatory mechanisms in alcoholic cardiomyopathy[J]. Biochim Biophys Acta Mol Basis Dis, 2018, 1864(6 Pt A): 2003-2009. DOI: 10.1016/j.bbadis.2018.03.016.
WARE J S, AMOR-SALAMANCA A, TAYAL U, et al . Genetic etiology for alcohol-induced cardiac toxicity[J ] . J Am Coll Cardiol, 2018, 71(20): 2293-2302. DOI: 10.1016/j.jacc. 2018.03.462.
TALMAN V, RUSKOAHO H. Cardiac fibrosis in myocardial infarction-from repair and remodeling to regeneration[J]. Cell Tissue Res, 2016, 365(3): 563-581. DOI: 10.1007/s00441-016-2431-9.
HEO S C, KIM Y N, CHOI Y, et al . Elevated expression of cathepsin K in periodontal ligament fibroblast by inflammatory cytokines accelerates osteoclastogenesis via paracrine mechanism in periodontal disease[J ] . Int J Mol Sci, 2021, 22(2): 695. DOI: 10.3390/ijms22020695..
FANG W Q, HE A N, XIANG M X, et al . Cathepsin K-deficiency impairs mouse cardiac function after myocardial infarction[J ] . J Mol Cell Cardiol, 2019, 127: 44-56. DOI: 10.1016/j.yjmcc.2018.11.010.
GUO R, HUA Y N, ROGERS O, et al . Cathepsin K knockout protects against cardiac dysfunction in diabetic mice[J ] . Sci Rep, 2017, 7(1): 8703. DOI: 10.1038/s41598-017-09037-z.
廉虹, 刘立会, 聂宇. 液质饲料喂养制备酒精性心肌病小鼠模型[J]. 中国比较医学杂志, 2020, 30(5): 40-46. DOI: 10.3969/j.issn.1671-7856.2020.05.007.
LIAN H, LIU L H, NIE Y. Establishment of a mouse model of alcoholic cardiomyopathy with liquid feed[J]. Chin J Comp Med, 2020, 30(5): 40-46. DOI: 10.3969/j.issn.1671-7856.2020.05.007.
GAO W Q, PAN B, LIU L J, et al . Alcohol exposure increases the expression of cardiac transcription factors through ERK1/2-mediated histone3 hyperacetylation in H9c2 cells[J ] . Biochem Biophys Res Commun, 2015, 466(4): 670-675. DOI: 10.1016/j.bbrc.2015.09.090.
卢薇, 王宁夫. 酒精性心肌损伤的临床分类及其临床特点分析[J]. 中华医学杂志, 2019, 99(41): 3237-3242. DOI: 10.3760/cma.j.issn.0376-2491.2019.41.008.
LU W, WANG N F. Clinical classification and characteristics of alcoholic myocardial injury[J]. Natl Med J China, 2019, 99(41): 3237-3242. DOI: 10.3760/cma.j.issn.0376-2491.2019.41.008.
LIU C L, GUO J L, ZHANG X, et al . Cysteine protease cathepsins in cardiovascular disease: from basic research to clinical trials[J ] . Nat Rev Cardiol, 2018, 15(6): 351-370. DOI: 10.1038/s41569-018-0002-3.
MCGLINCHEY R P, LACY S M, WALKER R L 3rd, et al . Cathepsin K is a potent disaggregase of α-synuclein fibrils[J ] . Biochem Biophys Res Commun, 2020, 529(4): 1106-1111. DOI: 10.1016/j.bbrc.2020.06.155.
SUN J S, SUKHOVA G K, ZHANG J, et al . Cathepsin K deficiency reduces elastase perfusion-induced abdominal aortic aneurysms in mice[J ] . Arterioscler Thromb Vasc Biol, 2012, 32(1): 15-23. DOI: 10.1161/ATVBAHA.111.235002.
路倩, 赵唯安, 刘玲娟, 等. 父代饮酒致子代小鼠心脏发育异常的组蛋白乙酰化修饰机制[J]. 中国病理生理杂志, 2019, 35(6): 975-980. DOI: 10.3969/j.issn.1000-4718.2019.06.003.
LU Q, ZHAO W A, LIU L J, et al . Mechanisms of histone acetylation modification in paternal alcohol consumption-induced heart development abnormalities of offspring[J ] . Chin J Pathophysiol, 2019, 35(6): 975-980. DOI: 10.3969/j.issn.1000-4718.2019.06.003.
尹玉洁, 张倩, 旷湘楠, 等. 内皮间质转分化在心肌纤维化中的研究进展[J]. 中国药理学通报, 2019, 35(1): 12-16. DOI: 10.3969/j.issn.1001-1978.2019.01.004.
YIN Y J, ZHANG Q, KUANG X N, et al . Research progress of endothelial mesenchymal transition in myocardial fibrosis[J ] . Chin Pharmacol Bull, 2019, 35(1): 12-16. DOI: 10.3969/j.issn.1001-1978.2019.01.004.
员小利, 井海云, 王丹. 丹参酮ⅡA通过抑制NADPH氧化酶保护心衰大鼠心肌纤维化的机制[J]. 安徽医科大学学报, 2019, 54(4): 600-604. DOI: 10.19405/j.cnki.issn1000-1492.2019.04.020.
YUAN X L, JING H Y, WANG D. Tanshinone ⅡA protects myocardial fibrosis in rats with heart failure by inhibiting NADPH oxidase[J]. Acta Univ Med Anhui, 2019, 54(4): 600-604. DOI: 10.19405/j.cnki.issn1000-1492.2019.04.020.
KAMIHARA T, BANDO Y K, MUROHARA T. Cardiac aging is regulated by autophagic disorder mediated by chronic inflammation via the Rubicon/Nox4 pathway[J ] . Eur Heart J, 2020, 41(Supplement_2): ehaa946.0842. DOI: 10.1093/ehjci/ehaa946.0842.
TAO Y L, ZHOU H, HUANG L L, et al . Schisandrin B protects against acute ethanol-induced cardiac injury by downregulating autophagy via the Nox4/ROS pathway[J ] . Pharmacology, 2021, 106(3/4): 177-188. DOI:10.1159/000510863..
SONG H, REN J. Protocatechuic acid attenuates angiotensin Ⅱ-induced cardiac fibrosis in cardiac fibroblasts through inhibiting the Nox4/ROS/p38 signaling pathway[J]. Phytother Res, 2019, 33(9): 2440-2447. DOI: 10.1002/ptr.6435.
PAN L L, LIU X H, SHEN Y Q, et al . Inhibition of NADPH oxidase 4-related signalingby sodium hydrosulfide attenuates myocardial fibrotic response[J ] . Int J Cardiol, 2013, 168(4): 3770-3778. DOI: 10.1016/j.ijcard.2013.06.007.
MROUEH F M, NOURELDEIN M, ZEIDAN Y H, et al . Unmasking the interplay between mTOR and Nox4: novel insights into the mechanism connecting diabetes and cancer[J ] . FASEB J, 2019, 33(12): 14051-14066. DOI:10.1096/fj.201900396RR.
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
