MicroRNA‑21‑5p acts via the PTEN/Akt/FOXO3a signaling pathway to prevent cardiomyocyte injury caused by high glucose/high fat conditions

Han,Ying; Cai,Xiaoqin; Pan,Min; Gong,Jin; Cai,Wenqin; Lu,Dan; Xu,Changsheng

doi:10.3892/etm.2022.11154

MicroRNA‑21‑5p acts via the PTEN/Akt/FOXO3a signaling pathway to prevent cardiomyocyte injury caused by high glucose/high fat conditions

Authors:
- Ying Han
- Xiaoqin Cai
- Min Pan
- Jin Gong
- Wenqin Cai
- Dan Lu
- Changsheng Xu
View Affiliations

Affiliations: Department of Geriatrics, The First Affiliated Hospital of Fujian Medical University, Fuzhou, Fujian 350005, P.R. China, Department of General Medicine, The First Affiliated Hospital of Fujian Medical University, Fuzhou, Fujian 350005, P.R. China, Fujian Institute of Hypertension, The First Affiliated Hospital of Fujian Medical University, Fuzhou, Fujian 350005, P.R. China
Published online on: January 18, 2022 https://doi.org/10.3892/etm.2022.11154
Article Number: 230
Copyright: © Han 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

MicroRNAs (miRNAs or miRs) play important roles in cardiovascular disease. miR‑21‑5p is known to be involved in the regulation of cardiomyocyte injury under high glucose and high fat (HG‑HF) conditions, but its mechanism of action remains unclear. In the present study, a cardiomyocyte cell line, H9c2, was treated with 33 mM glucose and 250 µM sodium palmitate for 24, 48, and 72 h to produce HG‑HF injury. After treatment, miR‑21‑5p expression was detected by reverse transcription‑quantitative PCR. A miR‑21‑5p mimic was then constructed and transfected into the cells and the potential molecular mechanism was investigated using Cell Counting Kit‑8, TUNEL, flow cytometry and western blot assays. Expression of miR‑21‑5p was significantly downregulated by HG‑HF treatment of H9c2 cells for 24, 48, and 72 h. In subsequent experiments, cells were treated for an intermediate period (48 h). Compared with the control group, HG‑HF treatment significantly inhibited H9c2 proliferation and promoted apoptosis, while these effects were significantly reduced in the miR‑21‑5p mimic. Compared with the control group, HG‑HF treatment significantly increased reactive oxygen species, while miR‑21‑5p mimic significantly reduced this effect. Compared with the control group, HG‑HF treatment significantly increased the expression of the pro‑apoptotic proteins Bax and phosphorylated (p)‑Akt and decreased the expression of the anti‑apoptotic proteins Bcl‑2, p‑PTEN, and p‑FOXO3a, while overexpression of miR‑21‑5p significantly reduced these effects. The results revealed that miR‑21‑5p inhibited apoptosis and oxidative stress in H9c2 cells induced by HG‑HF, likely through the PTEN/Akt/FOXO3a signaling pathway.

View Figures

View References

1	Picano E: Diabetic cardiomyopathy the importance of being earliest. J Am Coll Cardi-ol. 42:454–457. 2003.PubMed/NCBI View Article : Google Scholar
2	Avogaro A, Vigili de Kreutzenberg S, Negut C, Tiengo A and Scognamiglio R: Diabetic cardiomyopathy: A metabolic perspective. Am J Cardiol. 93:13A–16A. 2004.PubMed/NCBI View Article : Google Scholar
3	Zhang BB, Zhou G and Li C: AMPK: An emerging drug target for diabetes and the metabolic syndrome. Cell Metab. 9:407–416. 2009.PubMed/NCBI View Article : Google Scholar
4	Li B, Zheng Z, Wei Y, Wang M, Peng J, Kang T, Huang X, Xiao J, Li Y and Li Z: Therapeutic effects of neuregulin-1 in diabetic cardiomyopathy rats. Cardiovasc Diabetol. 10(69)2011.PubMed/NCBI View Article : Google Scholar
5	Wang Y, Zou L, Wu T, Xiong L, Zhang T, Kong L, Xue Y and Tang M: Identification of mRNA-miRNA crosstalk in human endothelial cells after exposure of PM2.5 through integrative transcriptome analysis. Ecotoxicol Environ Saf. 169:863–873. 2019.PubMed/NCBI View Article : Google Scholar
6	Yang J, Shi G, Gong Y, Cai J, Zheng Y and Zhang Z: LncRNA 0003250 accelerates heart autophagy and binds to miR-17-5p as a competitive endogenous RNA in chicken induced by selenium deficiency. J Cell Physiol. 236:157–177. 2021.PubMed/NCBI View Article : Google Scholar
7	Yang J, Gong Y, Cai J, Liu Q and Zhang Z: lnc-3215 Suppression Leads to Calcium Overload in Selenium Deficiency-Induced Chicken Heart Lesion via the lnc-3215-miR-1594-TNN2 Pathway. Mol Ther Nucleic Acids. 18:1–15. 2019.PubMed/NCBI View Article : Google Scholar
8	Song M, Zhao G, Sun H, Yao S, Zhou Z, Jiang P, Wu Q, Zhu H, Wang H, Dai C, et al: circPTPN12/miR-21-5 p/Np63alpha pathway contributes to human endometrial fibrosis. eLife 10: e65735.
9	Liu E, Lv L, Zhan Y, Ma Y, Feng J, He Y, Wen Y, Zhang Y, Pu Q, Ji F, et al: METTL3/N6-methyladenosine/ miR-21-5p promotes obstructive renal fibrosis by regulating inflammation through SPRY1/ERK/NF-κB pathway activation. J Cell Mol Med. 25:7660–7674. 2021.PubMed/NCBI View Article : Google Scholar
10	Qiao L, Hu S, Liu S, Zhang H, Ma H, Huang K, Li Z, Su T, Vandergriff A, Tang J, et al: microRNA-21-5p dysregulation in exosomes derived from heart failure patients impairs regenerative potential. J Clin Invest. 129:2237–2250. 2019.PubMed/NCBI View Article : Google Scholar
11	Raupach A, Torregroza C, Niestegge J, Feige K, Klemm-Meyer S, Bauer I, Brandenburger T, Grievink H, Heinen A and Huhn R: MiR-21-5p but not miR-1-3p expression is modulated by preconditioning in a rat model of myocardial infarction. Mol Biol Rep. 47:6669–6677. 2020.PubMed/NCBI View Article : Google Scholar
12	Dai B, Li H, Fan J, Zhao Y, Yin Z, Nie X, Wang DW and Chen C: MiR-21 protected against diabetic cardiomyopathy induced diastolic dysfunction by targeting gelsolin. Cardiovasc Diabetol. 17(123)2018.PubMed/NCBI View Article : Google Scholar
13	Balcells I, Cirera S and Busk PK: Specific and sensitive quantitative RT-PCR of miRNAs with DNA primers. BMC Biotechnol. 11(70)2011.PubMed/NCBI View Article : Google Scholar
14	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
15	Xia Y, Gong L, Liu H, Luo B, Li B, Li R, Li B, Lv M, Pan J and An F: Inhibition of prolyl hydroxylase 3 ameliorates cardiac dysfunction in diabetic cardiomyopathy. Mol Cell Endocrinol. 403:21–29. 2015.PubMed/NCBI View Article : Google Scholar
16	Wang CC and Reusch JE: Diabetes and cardiovascular disease: Changing the focus from glycemic control to improving long-term survival. Am J Cardiol. 110 (Suppl 9):58B–68B. 2012.PubMed/NCBI View Article : Google Scholar
17	Lee Y and Gustafsson AB: Role of apoptosis in cardiovascular disease. Apoptosis. 14:536–548. 2009.PubMed/NCBI View Article : Google Scholar
18	Shekhar A, Heeger P, Reutelingsperger C, Arbustini E, Narula N, Hofstra L, Bax JJ and Narula J: Targeted Imaging for Cell Death in Cardiovascular Disorders. JACC Cardiovasc Imaging. 11:476–493. 2018.PubMed/NCBI View Article : Google Scholar
19	Song Z, Shen F, Zhang Z, Wu S and Zhu G: Calpain inhibition ameliorates depression-like behaviors by reducing inflammation and promoting synaptic protein expression in the hippocampus. Neuropharmacology. 174(108175)2020.PubMed/NCBI View Article : Google Scholar
20	Song Z, Chen H, Xu W, Wu S and Zhu G: Basolateral amygdala calpain is required for extinction of contextual fear-memory. Neurobiol Learn Mem. 155:180–188. 2018.PubMed/NCBI View Article : Google Scholar
21	Myers MP, Stolarov JP, Eng C, Li J, Wang SI, Wigler MH, Parsons R and Tonks NK: P-TEN, the tumor suppressor from human chromosome 10q23, is a dual-specificity phosphatase. Proc Natl Acad Sci USA. 94:9052–9057. 1997.PubMed/NCBI View Article : Google Scholar
22	Shi YH, Wang YX, You JF, Heng WJ, Zhong HH and Fang WG: Activation of HIF-1 by bFGF in breast cancer: Role of PI-3K and MEK1/ERK pathways. Zhonghua Yi Xue Za Zhi. 84:1899–1903. 2004.PubMed/NCBI(In Chinese).
23	Song ZJ, Yang SJ, Han L, Wang B and Zhu G: Postnatal calpeptin treatment causes hippocampal neurodevelopmental defects in neonatal rats. Neural Regen Res. 14:834–840. 2019.PubMed/NCBI View Article : Google Scholar
24	Li D, Qu Y, Mao M, Zhang X, Li J, Ferriero D and Mu D: Involvement of the PTEN-AKT-FOXO3a pathway in neuronal apoptosis in developing rat brain after hypoxia-ischemia. J Cereb Blood Flow Metab. 29:1903–1913. 2009.PubMed/NCBI View Article : Google Scholar
25	Landgraf P, Rusu M, Sheridan R, Sewer A, Iovino N, Aravin A, Pfeffer S, Rice A, Kamphorst AO, Landthaler M, et al: A mammalian microRNA expression atlas based on small RNA library sequencing. Cell. 129:1401–1414. 2007.PubMed/NCBI View Article : Google Scholar
26	Sayed D, He M, Hong C, Gao S, Rane S, Yang Z and Abdellatif M: MicroRNA-21 is a downstream effector of AKT that mediates its antiapoptotic effects via suppression of Fas ligand. J Biol Chem. 285:20281–20290. 2010.PubMed/NCBI View Article : Google Scholar
27	Cheng Y, Liu X, Zhang S, Lin Y, Yang J and Zhang C: MicroRNA-21 protects against the H(2)O(2)-induced injury on cardiac myocytes via its target gene PDCD4. J Mol Cell Cardiol. 47:5–14. 2009.PubMed/NCBI View Article : Google Scholar
28	He Z, Long J, Yang C, Gong B, Cheng M, Wang Q and Tang J: LncRNA DGCR5 plays a tumor-suppressive role in glioma via the miR-21/Smad7 and miR-23a/PTEN axes. Aging (Albany NY). 12:20285–20307. 2020.PubMed/NCBI View Article : Google Scholar
29	Wang J, Yue J, Xia Q, Jiao X and Zhi J: Angiotensin II type i receptor agonistic autoantibodies induces apoptosis of cardiomyocytes by downregulating miR21 in preeclampsia: A mechanism study. Am J Transl Res. 11:2339–2349. 2019.PubMed/NCBI
30	Roy S, Khanna S, Hussain SR, Biswas S, Azad A, Rink C, Gnyawali S, Shilo S, Nuovo GJ and Sen CK: MicroRNA expression in response to murine myocardial infarction: miR-21 regulates fibroblast metalloprotease-2 via phosphatase and tensin homologue. Cardiovasc Res. 82:21–29. 2009.PubMed/NCBI View Article : Google Scholar
31	Meng F, Henson R, Wehbe-Janek H, Ghoshal K, Jacob ST and Patel T: MicroRNA-21 regulates expression of the PTEN tumor suppressor gene in human hepatocellular cancer. Gastroenterology. 133:647–658. 2007.PubMed/NCBI View Article : Google Scholar
32	Shen F, Song Z, Xie P, Li L, Wang B, Peng D and Zhu G: Polygonatum sibiricum polysaccharide prevents depression-like behaviors by reducing oxidative stress, inflammation, and cellular and synaptic damage. J Ethnopharmacol. 275(114164)2021.PubMed/NCBI View Article : Google Scholar
33	Qian M, Tan HM, Yu N, Wang T and Zhang Q: Inactivated Sendai Virus Induces ROS-dependent Apoptosis and Autophagy in Human Prostate Cancer Cells. Biomed Environ Sci. 31:280–289. 2018.PubMed/NCBI View Article : Google Scholar
34	Yang H, Xie Y, Yang D and Ren D: Oxidative stress-induced apoptosis in granulosa cells involves JNK, p53 and Puma. Oncotarget. 8:25310–25322. 2017.PubMed/NCBI View Article : Google Scholar
35	Sears CE, Ashley EA and Casadei B: Nitric oxide control of cardiac function: Is neuronal nitric oxide synthase a key component? Philos Trans R Soc Lond B Biol Sci. 359:1021–1044. 2004.PubMed/NCBI View Article : Google Scholar