Matrix metalloproteinase 3 regulates angiotensin II‑induced myocardial fibrosis cell viability, migration and apoptosis

Shu,Jin; Gu,Yiwen; Jin,Li; Wang,Haiya

doi:10.3892/mmr.2020.11790

Matrix metalloproteinase 3 regulates angiotensin II‑induced myocardial fibrosis cell viability, migration and apoptosis

Authors:
- Jin Shu
- Yiwen Gu
- Li Jin
- Haiya Wang
View Affiliations

Affiliations: Department of Gerontology, Shibei Hospital of Jing'an District, Shanghai 200443, P.R. China, Department of Gerontology, Shanghai Ninth People's Hospital, Shanghai JiaoTong University School of Medicine, Shanghai 200011, P.R. China
Published online on: December 20, 2020 https://doi.org/10.3892/mmr.2020.11790
Article Number: 151
Copyright: © Shu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Angiotensin II (AngII) is a central signaling molecule of the renin‑angiotensin system that serves a vital role in myocardial fibrosis (MF). The present study aimed to investigate the effects of matrix metalloproteinase (MMP)3 on MF progression. To induce cellular fibrosis, H9C2 rat myocardial cells were treated with AngII for 24 h. Subsequently, cells were treated with levocarnitine, or transfected with small interfering (si)RNA‑negative control or siRNA‑MMP3 (1/2/3). Cell viability, apoptosis and migration were assessed by performing Cell Counting Kit‑8, flow cytometry and Transwell assays, respectively. Reverse transcription‑quantitative PCR (RT‑qPCR) and western blotting were performed to determine the expression levels of MF biomarkers, including disease‑, apoptosis‑ and oxidative stress‑related genes. Compared with the control group, AngII significantly inhibited H9C2 cell viability and migration, and significantly increased H9C2 cell apoptosis (P<0.05). However, compared with AngII‑treated H9C2 cells, MMP3 knockdown significantly inhibited fibrotic H9C2 cell viability and migration, but increased fibrotic H9C2 cell apoptosis (P<0.05). The RT‑qPCR results demonstrated that MMP3 knockdown significantly downregulated the expression levels of AXL receptor tyrosine kinase, AngII receptor type 1, α‑smooth muscle actin and Collagen I in AngII‑treated H9C2 cells (P<0.05). Moreover, compared with AngII‑treated cells, MMP3 knockdown significantly decreased Bcl‑2 expression levels , but significantly increased caspase‑3 and p53 expression levels in AngII‑treated cells (P<0.05). Additionally, compared with AngII‑treated cells, MMP3 knockdown significantly decreased MMP3, MMP9, STAT3, p22Phox and p47Phox expression levels in AngII‑treated cells (P<0.05). The present study indicated that MMP3 knockdown altered myocardial fibroblast cell viability, migration and apoptosis by regulating apoptosis‑ and oxidative stress‑related genes, thus delaying MF progression.

View Figures

View References

1	Talman V and Ruskoaho H: Cardiac fibrosis in myocardial infarction-from repair and remodeling to regeneration. Cell Tissue Res. 365:563–581. 2016. View Article : Google Scholar : PubMed/NCBI
2	Hao G, Wang X, Chen Z, Zhang L, Zhang Y, Wei B, Zheng C, Kang Y, Jiang L, Zhu Z, et al China Hypertension Survey Investigators, : Prevalence of heart failure and left ventricular dysfunction in China: The China Hypertension Survey, 2012–2015. Eur J Heart Fail. 21:1329–1337. 2019. View Article : Google Scholar : PubMed/NCBI
3	González A, Schelbert EB, Díez J and Butler J: Myocardial interstitial fibrosis in heart failure: Biological and translational perspectives. J Am Coll Cardiol. 71:1696–1706. 2018. View Article : Google Scholar : PubMed/NCBI
4	Kong P, Christia P and Frangogiannis NG: The pathogenesis of cardiac fibrosis. Cell Mol Life Sci. 71:549–574. 2014. View Article : Google Scholar : PubMed/NCBI
5	Aoki T, Fukumoto Y, Sugimura K, Oikawa M, Satoh K, Nakano M, Nakayama M and Shimokawa H: Prognostic impact of myocardial interstitial fibrosis in non-ischemic heart failure. -Comparison between preserved and reduced ejection fraction heart failure. Circ J. 75:2605–2613. 2011. View Article : Google Scholar : PubMed/NCBI
6	Gyöngyösi M, Winkler J, Ramos I, Do QT, Firat H, McDonald K, González A, Thum T, Díez J, Jaisser F, et al: Myocardial fibrosis: Biomedical research from bench to bedside. Eur J Heart Fail. 19:177–191. 2017. View Article : Google Scholar : PubMed/NCBI
7	Helm PA, Caravan P, French BA, Jacques V, Shen L, Xu Y, Beyers RJ, Roy RJ, Kramer CM and Epstein FH: Postinfarction myocardial scarring in mice: Molecular MR imaging with use of a collagen-targeting contrast agent. Radiology. 247:788–796. 2008. View Article : Google Scholar : PubMed/NCBI
8	López B, González A, Ravassa S, Beaumont J, Moreno MU, San José G, Querejeta R and Díez J: Circulating biomarkers of myocardial fibrosis: The need for a reappraisal. J Am Coll Cardiol. 65:2449–2456. 2015. View Article : Google Scholar : PubMed/NCBI
9	Ellison DH and Felker GM: Diuretic treatment in heart failure. N Engl J Med. 377:1964–1975. 2017. View Article : Google Scholar : PubMed/NCBI
10	Schupp T, Behnes M, Weiss C, Nienaber C, Reiser L, Bollow A, Taton G, Reichelt T, Ellguth D, Engelke N, et al: Digitalis therapy and risk of recurrent ventricular tachyarrhythmias and ICD therapies in atrial fibrillation and heart failure. Cardiology. 142:129–140. 2019. View Article : Google Scholar : PubMed/NCBI
11	Schafer S, Viswanathan S, Widjaja AA, Lim WW, Moreno-Moral A, DeLaughter DM, Ng B, Patone G, Chow K, Khin E, et al: IL-11 is a crucial determinant of cardiovascular fibrosis. Nature. 552:110–115. 2017. View Article : Google Scholar : PubMed/NCBI
12	Shinde AV, Humeres C and Frangogiannis NG: The role of α-smooth muscle actin in fibroblast-mediated matrix contraction and remodeling. Biochim Biophys Acta Mol Basis Dis. 1863:298–309. 2017. View Article : Google Scholar : PubMed/NCBI
13	Zhang Y, Luo G, Zhang Y, Zhang M, Zhou J, Gao W, Xuan X, Yang X, Yang D, Tian Z, et al: Critical effects of long non-coding RNA on fibrosis diseases. Exp Mol Med. 50:e4282018. View Article : Google Scholar : PubMed/NCBI
14	Zhang X, Pan L, Yang K, Fu Y, Liu Y, Chi J, Zhang X, Hong S, Ma X and Yin X: H3 relaxin protects against myocardial injury in experimental diabetic cardiomyopathy by inhibiting myocardial apoptosis, fibrosis and inflammation. Cell Physiol Biochem. 43:1311–1324. 2017. View Article : Google Scholar : PubMed/NCBI
15	Kazakov A, Hall RA, Werner C, Meier T, Trouvain A, Rodionycheva S, Nickel A, Lammert F, Maack C, Böhm M, et al: Raf kinase inhibitor protein mediates myocardial fibrosis under conditions of enhanced myocardial oxidative stress. Basic Res Cardiol. 113:422018. View Article : Google Scholar : PubMed/NCBI
16	Batlle M, Recarte-Pelz P, Roig E, Castel MA, Cardona M, Farrero M, Ortiz JT, Campos B, Pulgarín MJ, Ramírez J, et al: AXL receptor tyrosine kinase is increased in patients with heart failure. Int J Cardiol. 173:402–409. 2014. View Article : Google Scholar : PubMed/NCBI
17	Zheng RH, Bai XJ, Zhang WW, Wang J, Bai F, Yan CP, James EA, Bose HS, Wang NP and Zhao ZQ: Liraglutide attenuates cardiac remodeling and improves heart function after abdominal aortic constriction through blocking angiotensin II type 1 receptor in rats. Drug Des Devel Ther. 13:2745–2757. 2019. View Article : Google Scholar : PubMed/NCBI
18	Zachariah JP, Colan SD, Lang P, Triedman JK, Alexander ME, Walsh EP, Berul CI and Cecchin F: Circulating matrix metalloproteinases in adolescents with hypertrophic cardiomyopathy and ventricular arrhythmia. Circ Heart Fail. 5:462–466. 2012. View Article : Google Scholar : PubMed/NCBI
19	Shu J, Liu Z, Jin L and Wang H: An RNA sequencing study identifies candidate genes for angiotensin II induced cardiac remodeling. Mol Med Rep. 17:1954–1962. 2018.PubMed/NCBI
20	Craig VJ, Zhang L, Hagood JS and Owen CA: Matrix metalloproteinases as therapeutic targets for idiopathic pulmonary fibrosis. Am J Respir Cell Mol Biol. 53:585–600. 2015. View Article : Google Scholar : PubMed/NCBI
21	Kitanaka N, Nakano R, Sakai M, Kitanaka T, Namba S, Konno T, Nakayama T and Sugiya H: ERK1/ATF-2 signaling axis contributes to interleukin-1β-induced MMP-3 expression in dermal fibroblasts. PLoS One. 14:e02228692019. View Article : Google Scholar : PubMed/NCBI
22	Tuncer T, Kaya A, Gulkesen A, Kal GA, Kaman D and Akgol G: Matrix metalloproteinase-3 levels in relation to disease activity and radiological progression in rheumatoid arthritis. Adv Clin Exp Med. 28:665–670. 2019. View Article : Google Scholar : PubMed/NCBI
23	Patel VB, Zhong JC, Grant MB and Oudit GY: Role of the ACE2/Angiotensin 1–7 axis of the renin-angiotensin system in heart failure. Circ Res. 118:1313–1326. 2016. View Article : Google Scholar : PubMed/NCBI
24	Jin HY, Song B, Oudit GY, Davidge ST, Yu HM, Jiang YY, Gao PJ, Zhu DL, Ning G, Kassiri Z, et al: ACE2 deficiency enhances angiotensin II-mediated aortic profilin-1 expression, inflammation and peroxynitrite production. PLoS One. 7:e385022012. View Article : Google Scholar : PubMed/NCBI
25	Sahoo S, Meijles DN, Al Ghouleh I, Tandon M, Cifuentes-Pagano E, Sembrat J, Rojas M, Goncharova E and Pagano PJ: MEF2C-MYOCD and leiomodin1 suppression by miRNA-214 promotes smooth muscle cell phenotype switching in pulmonary arterial hypertension. PLoS One. 11:e01537802016. View Article : Google Scholar : PubMed/NCBI
26	Xu G, Ao R, Zhi Z, Jia J and Yu B: miR-21 and miR-19b delivered by hMSC-derived EVs regulate the apoptosis and differentiation of neurons in patients with spinal cord injury. J Cell Physiol. 234:10205–10217. 2019. View Article : Google Scholar : PubMed/NCBI
27	Fagone P, Mangano K, Pesce A, Portale TR, Puleo S and Nicoletti F: Emerging therapeutic targets for the treatment of hepatic fibrosis. Drug Discov Today. 21:369–375. 2016. View Article : Google Scholar : PubMed/NCBI
28	Gungor O, Unal HU, Guclu A, Gezer M, Eyileten T, Guzel FB, Altunoren O, Erken E, Oguz Y, Kocyigit I, et al: IL-33 and ST2 levels in chronic kidney disease: Associations with inflammation, vascular abnormalities, cardiovascular events, and survival. PLoS One. 12:e01789392017. View Article : Google Scholar : PubMed/NCBI
29	Ke B, Zhu N, Luo F, Xu Y and Fang X: Targeted inhibition of endoplasmic reticulum stress: New hope for renal fibrosis (Review). Mol Med Rep. 16:1014–1020. 2017. View Article : Google Scholar : PubMed/NCBI
30	Fagone P, Mangano K, Mammana S, Pesce A, Pesce A, Caltabiano R, Giorlandino A, Portale TR, Cavalli E, Lombardo GA, et al: Identification of novel targets for the diagnosis and treatment of liver fibrosis. Int J Mol Med. 36:747–752. 2015. View Article : Google Scholar : PubMed/NCBI
31	Ambale-Venkatesh B, Liu CY, Liu YC, Donekal S, Ohyama Y, Sharma RK, Wu CO, Post WS, Hundley GW, Bluemke DA, et al: Association of myocardial fibrosis and cardiovascular events: The multi-ethnic study of atherosclerosis. Eur Heart J Cardiovasc Imaging. 20:168–176. 2019. View Article : Google Scholar : PubMed/NCBI
32	Chin CWL, Everett RJ, Kwiecinski J, Vesey AT, Yeung E, Esson G, Jenkins W, Koo M, Mirsadraee S, White AC, et al: Myocardial fibrosis and cardiac decompensation in aortic stenosis. JACC Cardiovasc Imaging. 10:1320–1333. 2017. View Article : Google Scholar : PubMed/NCBI
33	Gulati A, Japp AG, Raza S, Halliday BP, Jones DA, Newsome S, Ismail NA, Morarji K, Khwaja J, Spath N, et al: Absence of myocardial fibrosis predicts favorable long-term survival in new-onset heart failure. Circ Cardiovasc Imaging. 11:e0077222018. View Article : Google Scholar : PubMed/NCBI
34	Mirastschijski U, Lupše B, Maedler K, Sarma B, Radtke A, Belge G, Dorsch M, Wedekind D, McCawley LJ, Boehm G, et al: Matrix metalloproteinase-3 is key effector of TNF-α-induced collagen degradation in skin. Int J Mol Sci. 20:202019. View Article : Google Scholar
35	Wang Q, Sui X, Chen R, Ma PY, Teng YL, Ding T, Sui DJ and Yang P: Ghrelin ameliorates angiotensin ii-induced myocardial fibrosis by upregulating peroxisome proliferator-activated receptor gamma in young male rats. BioMed Res Int. 2018:98975812018.PubMed/NCBI
36	Wu P, Liu Z, Zhao T, Xia F, Gong L, Zheng Z, Chen Z, Yang T and Duan Q: Lovastatin attenuates angiotensin II induced cardiovascular fibrosis through the suppression of YAP/TAZ signaling. Biochem Biophys Res Commun. 512:736–741. 2019. View Article : Google Scholar : PubMed/NCBI
37	Berg G, Barchuk M and Miksztowicz V: Behavior of metalloproteinases in adipose tissue, liver and arterial wall: An update of extracellular matrix remodeling. Cells. 8:82019. View Article : Google Scholar
38	Taha EA, Sogawa C, Okusha Y, Kawai H, Oo MW, Elseoudi A, Lu Y, Nagatsuka H, Kubota S, Satoh A, et al: Knockout of MMP3 weakens solid tumor organoids and cancer extracellular vesicles. Cancers (Basel). 12:122020. View Article : Google Scholar
39	Bufu T, Di X, Yilin Z, Gege L, Xi C and Ling W: Celastrol inhibits colorectal cancer cell proliferation and migration through suppression of MMP3 and MMP7 by the PI3K/AKT signaling pathway. Anticancer Drugs. 29:530–538. 2018. View Article : Google Scholar : PubMed/NCBI
40	McShane L, Tabas I, Lemke G, Kurowska-Stolarska M and Maffia P: TAM receptors in cardiovascular disease. Cardiovasc Res. 115:1286–1295. 2019. View Article : Google Scholar : PubMed/NCBI
41	Zhang LH, Pang XF, Bai F, Wang NP, Shah AI, McKallip RJ, Li XW, Wang X and Zhao ZQ: Preservation of glucagon-like peptide-1 level attenuates angiotensin ii-induced tissue fibrosis by altering AT1/AT2 receptor expression and angiotensin-converting enzyme 2 activity in rat heart. Cardiovasc Drugs Ther. 29:243–255. 2015. View Article : Google Scholar : PubMed/NCBI
42	Zhang WW, Bai F, Wang J, Zheng RH, Yang LW, James EA and Zhao ZQ: Edaravone inhibits pressure overload-induced cardiac fibrosis and dysfunction by reducing expression of angiotensin II AT1 receptor. Drug Des Devel Ther. 11:3019–3033. 2017. View Article : Google Scholar : PubMed/NCBI
43	Harikrishnan V, Titus AS, Cowling RT and Kailasam S: Collagen receptor cross-talk determines α-smooth muscle actin-dependent collagen gene expression in angiotensin II-stimulated cardiac fibroblasts. J Biol Chem. 294:19723–19739. 2019. View Article : Google Scholar : PubMed/NCBI
44	Li N, Feng F, Wu K, Zhang H, Zhang W and Wang W: Inhibitory effects of astragaloside IV on silica-induced pulmonary fibrosis via inactivating TGF-beta1/Smad3 signaling. Biomed Pharmacother. 119:1093872019. View Article : Google Scholar : PubMed/NCBI
45	Weder B, Mamie C, Rogler G, Clarke S, McRae B, Ruiz PA and Hausmann M: BCL2 regulates differentiation of intestinal fibroblasts. Inflamm Bowel Dis. 24:1953–1966. 2018. View Article : Google Scholar : PubMed/NCBI
46	Lossi L, Castagna C and Merighi A: Caspase-3 mediated cell death in the normal development of the mammalian cerebellum. Int J Mol Sci. 19:192018. View Article : Google Scholar
47	Wang X, Simpson ER and Brown KA: p53: Protection against tumor growth beyond effects on cell cycle and apoptosis. Cancer Res. 75:5001–5007. 2015. View Article : Google Scholar : PubMed/NCBI
48	Wang JH, Zhao L, Pan X, Chen NN, Chen J, Gong QL, Su F, Yan J, Zhang Y and Zhang SH: Hypoxia-stimulated cardiac fibroblast production of IL-6 promotes myocardial fibrosis via the TGF-β1 signaling pathway. Lab Invest. 96:839–852. 2016. View Article : Google Scholar : PubMed/NCBI
49	Yao C, Cao X, Fu Z, Tian J, Dong W, Xu J, An K, Zhai L and Yu J: Boschniakia rossica polysaccharide triggers laryngeal carcinoma cell apoptosis by regulating expression of Bcl-2, Caspase-3, and P53. Med Sci Monit. 23:2059–2064. 2017. View Article : Google Scholar : PubMed/NCBI
50	Tsutsui H, Kinugawa S and Matsushima S: Oxidative stress and heart failure. Am J Physiol Heart Circ Physiol. 301:H2181–H2190. 2011. View Article : Google Scholar : PubMed/NCBI
51	Singh S and Torzewski M: Fibroblasts and their pathological functions in the fibrosis of aortic valve sclerosis and atherosclerosis. Biomolecules. 9:92019. View Article : Google Scholar
52	Wang M, Kim SH, Monticone RE and Lakatta EG: Matrix metalloproteinases promote arterial remodeling in aging, hypertension, and atherosclerosis. Hypertension. 65:698–703. 2015. View Article : Google Scholar : PubMed/NCBI
53	Zhang M, Xue Y, Chen H, Meng L, Chen B, Gong H, Zhao Y and Qi R: Resveratrol inhibits MMP3 and MMP9 expression and secretion by suppressing TLR4/NF-κB/STAT3 activation in Ox-LDL-treated HUVECs. Oxid Med Cell Longev. 2019:90131692019.PubMed/NCBI
54	Szczepanek K, Chen Q, Larner AC and Lesnefsky EJ: Cytoprotection by the modulation of mitochondrial electron transport chain: The emerging role of mitochondrial STAT3. Mitochondrion. 12:180–189. 2012. View Article : Google Scholar : PubMed/NCBI
55	Han J, Ye S, Zou C, Chen T, Wang J, Li J, Jiang L, Xu J, Huang W, Wang Y, et al: Angiotensin II causes biphasic STAT3 activation through TLR4 to initiate cardiac remodeling. Hypertension. 72:1301–1311. 2018. View Article : Google Scholar : PubMed/NCBI
56	Yim J, Cho H and Rabkin SW: Gene expression and gene associations during the development of heart failure with preserved ejection fraction in the Dahl salt sensitive model of hypertension. Clin Exp Hypertens. 40:155–166. 2018. View Article : Google Scholar : PubMed/NCBI