Identification of miR‑145 as a regulator of the cardiomyocyte inflammatory response and oxidative stress under hyperglycemia

Zheng,Wan; Li,Tianfa; Wei,Junping; Zhang,Yuanyuan; Zuo,Qi; Lin,Yun

doi:10.3892/etm.2021.9898

Identification of miR‑145 as a regulator of the cardiomyocyte inflammatory response and oxidative stress under hyperglycemia

Authors:
- Wan Zheng
- Tianfa Li
- Junping Wei
- Yuanyuan Zhang
- Qi Zuo
- Yun Lin
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Affiliations: Department of Cardiovascular Internal Medicine, The First Affiliated Hospital of Hainan Medical University, Haikou, Hainan 570102, P.R. China
Published online on: March 8, 2021 https://doi.org/10.3892/etm.2021.9898
Article Number: 467
Copyright: © Zheng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The current study aimed to explore the effects of microRNA (miR)‑145 on the inflammatory response and oxidative stress (OS) in high glucose (HG)‑induced cardiomyocytes, as well as the specific mechanism underlying this action. H9c2 cells were treated with 33 mmol/l glucose (HG group) or cotreated with 24.5 mmol/l mannitol and 5.5 mmol/l glucose (hypertonic group), and the expression levels of miR‑145 and ADP ribosylation factor 6 (ARF6) were detected. The cells were transfected with pcDNA3.1‑ARF6, miR‑145 mimics or corresponding negative controls prior to the assessment of cell survival rate. Levels of lactate dehydrogenase (LDH), reactive oxygen species (ROS) and malondialdehyde (MDA), as well as the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx), and the levels of IL‑6, TNF‑α and monocyte chemoattractant protein‑1 (MCP‑1) were subsequently determined. The apoptotic rate of H9c2 cells was examined by flow cytometry. The interaction between miR‑145‑ARF6 was predicted and confirmed by luciferase reporter assays. In the HG group, miR‑145 expression was significantly decreased and ARF6 expression significantly increased compared with controls. Furthermore, the levels of inflammatory factors (IL‑6, TNF‑α and MCP‑1), LDH, ROS and MDA were significantly elevated in the HG group compared with controls. Significantly decreased SOD, CAT and GPx activities and significantly increased numbers of apoptotic cells were observed in the HG group compared with controls. The cells transfected with miR‑145 mimics exhibited significantly decreased LDH, ROS and MDA levels, significantly increased antioxidant enzyme activities and significantly decreased apoptotic rates compared with controls, while the opposite results were observed in cells transfected with pcDNA3.1‑ARF6. Moreover, co‑transfection with miR‑145 mimics and pcDNA3.1‑ARF6 exacerbated the inflammatory response and OS injury in HG‑induced cardiomyocytes compared with cells transfected with miR‑145 mimics alone. Furthermore, miR‑145 negatively targeted ARF6. miR‑145 attenuated the HG‑induced inflammatory response and OS injury in cardiomyocytes by negatively regulating ARF6, which may contribute to providing a theoretical basis for the treatment of diabetic cardiomyopathy.

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1	Newman JD, Vani AK, Aleman JO, Weintraub HS, Berger JS and Schwartzbard AZ: The changing landscape of diabetes therapy for cardiovascular risk reduction: JACC state-of-the-art review. J Am Coll Cardiol. 72:1856–1869. 2018.PubMed/NCBI View Article : Google Scholar
2	Jia G, Whaley-Connell A and Sowers JR: Diabetic cardiomyopathy: A hyperglycaemia- and insulin-resistance-induced heart disease. Diabetologia. 61:21–28. 2018.PubMed/NCBI View Article : Google Scholar
3	Bugger H and Abel ED: Molecular mechanisms of diabetic cardiomyopathy. Diabetologia. 57:660–671. 2014.PubMed/NCBI View Article : Google Scholar
4	Karam BS, Chavez-Moreno A, Koh W, Akar JG and Akar FG: Oxidative stress and inflammation as central mediators of atrial fibrillation in obesity and diabetes. Cardiovasc Diabetol. 16(120)2017.PubMed/NCBI View Article : Google Scholar
5	Newsholme P, Cruzat VF, Keane KN, Carlessi R and de Bittencourt PI Jr: Molecular mechanisms of ROS production and oxidative stress in diabetes. Biochem J. 473:4527–4550. 2016.PubMed/NCBI View Article : Google Scholar
6	Qadir MMF, Klein D, Alvarez-Cubela S, Dominguez-Bendala J and Pastori RL: The role of MicroRNAs in diabetes-related oxidative stress. Int J Mol Sci. 20(5423)2019.PubMed/NCBI View Article : Google Scholar
7	Kyrychenko S, Kyrychenko V, Badr MA, Ikeda Y, Sadoshima J and Shirokova N: Pivotal role of miR-448 in the development of ROS-induced cardiomyopathy. Cardiovasc Res. 108:324–334. 2015.PubMed/NCBI View Article : Google Scholar
8	Tang Q, Li MY, Su YF, Fu J, Zou ZY, Wang Y and Li SN: Absence of miR-223-3p ameliorates hypoxia-induced injury through repressing cardiomyocyte apoptosis and oxidative stress by targeting KLF15. Eur J Pharmacol. 841:67–74. 2018.PubMed/NCBI View Article : Google Scholar
9	Su Q, Yao J and Sheng C: Geniposide attenuates LPS-induced injury via up-regulation of miR-145 in H9c2 cells. Inflammation. 41:1229–1237. 2018.PubMed/NCBI View Article : Google Scholar
10	Li L, Liu M, He L, Wang S and Cui S: Baicalin relieves TNF-α-evoked injury in human aortic endothelial cells by up-regulation of miR-145. Phytother Res. 34:836–845. 2020.PubMed/NCBI View Article : Google Scholar
11	He M, Wu N, Leong MC, Zhang W, Ye Z, Li R, Huang J, Zhang Z, Li L, Yao X, et al: miR-145 improves metabolic inflammatory disease through multiple pathways. J Mol Cell Biol. 12:152–162. 2020.PubMed/NCBI View Article : Google Scholar
12	Yuan M, Zhang L, You F, Zhou J, Ma Y, Yang F and Tao L: MiR-145-5p regulates hypoxia-induced inflammatory response and apoptosis in cardiomyocytes by targeting CD40. Mol Cell Biochem. 431:123–131. 2017.PubMed/NCBI View Article : Google Scholar
13	Hui Y and Yin Y: MicroRNA-145 attenuates high glucose-induced oxidative stress and inflammation in retinal endothelial cells through regulating TLR4/NF-κB signaling. Life Sci. 207:212–218. 2018.PubMed/NCBI View Article : Google Scholar
14	Padival AK, Hawkins KS and Huang C: High glucose-induced membrane translocation of PKC betaI is associated with Arf6 in glomerular mesangial cells. Mol Cell Biochem. 258:129–135. 2004.PubMed/NCBI View Article : Google Scholar
15	Fang Z, Miao Y, Ding X, Deng H, Liu S, Wang F, Zhou R, Watson C, Fu C, Hu Q, et al: Proteomic identification and functional characterization of a novel ARF6 GTPase-activating protein, ACAP4. Mol Cell Proteomics. 5:1437–1449. 2006.PubMed/NCBI View Article : Google Scholar
16	Karnik R, Ludlow MJ, Abuarab N, Smith AJ, Hardy ME, Elliott DJ and Sivaprasadarao A: Endocytosis of HERG is clathrin-independent and involves arf6. PLoS One. 8(e85630)2013.PubMed/NCBI View Article : Google Scholar
17	Li R, Shen Q, Wu N, He M, Liu N, Huang J, Lu B, Yao Q, Yang Y and Hu R: MiR-145 improves macrophage-mediated inflammation through targeting Arf6. Endocrine. 60:73–82. 2018.PubMed/NCBI View Article : Google Scholar
18	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001.PubMed/NCBI View Article : Google Scholar
19	Gilca GE, Stefanescu G, Badulescu O, Tanase DM, Bararu I and Ciocoiu M: Diabetic cardiomyopathy: Current approach and potential diagnostic and therapeutic targets. J Diabetes Res. 2017(1310265)2017.PubMed/NCBI View Article : Google Scholar
20	Li H, Fan J, Zhao Y, Zhang X, Dai B, Zhan J, Yin Z, Nie X, Fu XD, Chen C and Wang DW: Nuclear miR-320 mediates diabetes-induced cardiac dysfunction by activating transcription of fatty acid metabolic genes to cause lipotoxicity in the heart. Circ Res. 125:1106–1120. 2019.PubMed/NCBI View Article : Google Scholar
21	Wang Y, Zhao RZ, Chen PK, Xu GX, Liu ZJ, Long XP, Qiu ZM and Shi B: Impact and related mechanism on the improvement of hyperglycemia-induced pyroptosis in H9c2 cells by mircoRNA-214. Zhonghua Xin Xue Guan Bing Za Zhi. 47:820–828. 2019.PubMed/NCBI View Article : Google Scholar : (In Chinese).
22	Yang X, Li X, Lin Q and Xu Q: Up-regulation of microRNA-203 inhibits myocardial fibrosis and oxidative stress in mice with diabetic cardiomyopathy through the inhibition of PI3K/Akt signaling pathway via PIK3CA. Gene. 715(143995)2019.PubMed/NCBI View Article : Google Scholar
23	Zhong F, Xu J, Yang X, Zhang Q, Gao Z, Deng Y, Zhang L and Yu C: miR-145 eliminates lipopolysaccharides-induced inflammatory injury in human fibroblast-like synoviocyte MH7A cells. J Cell Biochem. 119:10059–10066. 2018.PubMed/NCBI View Article : Google Scholar
24	Shi J, Jiang K and Li Z: MiR-145 ameliorates neuropathic pain via inhibiting inflammatory responses and mTOR signaling pathway by targeting Akt3 in a rat model. Neurosci Res. 134:10–17. 2018.PubMed/NCBI View Article : Google Scholar
25	Lin H, You B, Lin X, Wang X, Zhou D, Chen Z, Chen Y and Wang R: Silencing of long non-coding RNA Sox2ot inhibits oxidative stress and inflammation of vascular smooth muscle cells in abdominal aortic aneurysm via microRNA-145-mediated Egr1 inhibition. Aging (Albany NY). 12:12684–12702. 2020.PubMed/NCBI View Article : Google Scholar
26	Abdul-Salam VB, Russomanno G, Chien-Nien C, Mahomed AS, Yates LA, Wilkins MR, Zhao L, Gierula M, Dubois O, Schaeper U, et al: CLIC4/Arf6 pathway. Circ Res. 124:52–65. 2019.PubMed/NCBI View Article : Google Scholar
27	Onodera Y, Nam JM, Horikawa M, Shirato H and Sabe H: Arf6-driven cell invasion is intrinsically linked to TRAK1-mediated mitochondrial anterograde trafficking to avoid oxidative catastrophe. Nat Commun. 9(2682)2018.PubMed/NCBI View Article : Google Scholar
28	Jayaram B and Kowluru A: Phagocytic NADPH oxidase links ARNO-Arf6 signaling pathway in glucose-stimulated insulin secretion from the pancreatic β-cell. Cell Physiol Biochem. 30:1351–1362. 2012.PubMed/NCBI View Article : Google Scholar
29	Li C, Zhang J, Xue M, Li X, Han F, Liu X, Xu L, Lu Y, Cheng Y, Li T, et al: SGLT2 inhibition with empagliflozin attenuates myocardial oxidative stress and fibrosis in diabetic mice heart. Cardiovasc Diabetol. 18(15)2019.PubMed/NCBI View Article : Google Scholar
30	Ke Y, Wang C, Zhang J, Zhong X, Wang R, Zeng X and Ba X: The role of PARPs in inflammation-and metabolic-related diseases: Molecular mechanisms and beyond. Cells. 8(1047)2019.PubMed/NCBI View Article : Google Scholar
31	Fehr AR, Singh SA, Kerr CM, Mukai S, Higashi H and Aikawa M: The impact of PARPs and ADP-ribosylation on inflammation and host-pathogen interactions. Genes Dev. 34:341–359. 2020.PubMed/NCBI View Article : Google Scholar
32	Dani N, Barbosa AJ, Del Rio A and Di Girolamo M: ADP-ribosylated proteins as old and new drug targets for anticancer therapy: The example of ARF6. Curr Pharm Des. 19:624–633. 2013.PubMed/NCBI
33	Ren C, Yuan Q, Jian X, Randazzo PA, Tang W and Wu D: Small GTPase ARF6 is a coincidence-detection code for RPH3A polarization in neutrophil polarization. J Immunol. 204:1012–1021. 2020.PubMed/NCBI View Article : Google Scholar
34	Eades G, Wolfson B, Zhang Y, Li Q, Yao Y and Zhou Q: lincRNA-RoR and miR-145 regulate invasion in triple-negative breast cancer via targeting ARF6. Mol Cancer Res. 13:330–338. 2015.PubMed/NCBI View Article : Google Scholar
35	Hsu WC, Li WM, Lee YC, Huang AM, Chang LL, Lin HH, Wu WJ, Li CC, Liang PI and Ke HL: MicroRNA-145 suppresses cell migration and invasion in upper tract urothelial carcinoma by targeting ARF6. FASEB J. 34:5975–5992. 2020.PubMed/NCBI View Article : Google Scholar