miR‑155 inhibition represents a potential valuable regulator in mitigating myocardial hypoxia/reoxygenation injury through targeting BAG5 and MAPK/JNK signaling

Xi,Jing; Li,Qiang‑Qiang; Li,Bing‑Qiang; Li,Ning

doi:10.3892/mmr.2020.10924

miR‑155 inhibition represents a potential valuable regulator in mitigating myocardial hypoxia/reoxygenation injury through targeting BAG5 and MAPK/JNK signaling

Authors:
- Jing Xi
- Qiang‑Qiang Li
- Bing‑Qiang Li
- Ning Li
View Affiliations

Affiliations: Department of Cardiology, Anqiu People's Hospital, Weifang, Shandong 262100, P.R. China, Department of Cardiology in Integrated Traditional Chinese and Western Medicine, Anqiu People's Hospital, Weifang, Shandong 262100, P.R. China
Published online on: January 9, 2020 https://doi.org/10.3892/mmr.2020.10924
Pages: 1011-1020
Copyright: © Xi 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

Increasing evidence has indicated that miR‑155 is closely associated with apoptosis, which may protect the myocardium and diminish the infarct area in myocardial ischemia reperfusion injury (IRI). In addition, studies have revealed that miR‑155 serves a leading role in promoting fibroblast inflammation, cardiac dysfunction and other aspects of myocardial injury. The present study aimed to uncover the function and potential biological mechanism of miR‑155 in myocardial IRI. The rat H9c2 myocardial cells was treated with hypoxia/reoxygenation (H/R) to simulate IRI in vitro. Reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) was used to detect the expression levels of miR‑155 mRNA. Cell Counting Kit‑8 and flow cytometry assays and western blot analysis were applied to determine the biological behaviors of the H/R‑treated cells. The association between miR‑155 and BAG family molecular chaperone regulator 5 (BAG5) was predicted by bioinformatics software and was confirmed by dual luciferase assay. RT‑qPCR and western blot analysis were used to analyze the expression of BAG5. The key proteins involved in mitogen‑activated protein kinase (MAPK)/JNK signaling pathway were detected by western blot analysis. The data from the RT‑qPCR assay indicated that the expression of miR‑155 was markedly upregulated in the H/R model, and that downregulation of miR‑155 may promote cell proliferation and inhibit cell apoptosis, and vice versa. BAG5, which was downregulated in the H/R model, was confirmed as a target of miR‑155 and negatively modulated by miR‑155. The key proteins involved in MAPK/JNK signaling, which were highly expressed in the H/R model, were suppressed by treatment with the miR‑155 inhibitor, and overexpression of BAG5 promoted the protective effect of miR‑155 inhibition on cell injury caused by H/R. In addition, the expression patterns of hypoxia‑inducible factor 1‑α and von Hippel‑Lindau were altered following different treatments. Taken together, the data from the present study indicated that miR‑155 inhibition represented a potential treatment strategy to improve myocardial H/R injury, which may be associated with targeting BAG5 and inhibition of the MAPK/JNK pathway.

View Figures

View References

1	Jennings RB, Sommers HM, Smyth GA, Flack HA and Linn H: Myocardial necrosis induced by temporary occlusion of a coronary artery in the dog. Arch Pathol. 70:68–78. 1960.PubMed/NCBI
2	Eltzschig HK and Eckle T: Ischemia and reperfusion-from mechanism to translation. Nat Med. 17:1391–401. 2011. View Article : Google Scholar : PubMed/NCBI
3	Sun G, Lu Y, Li Y, Mao J, Zhang J, Jin Y, Li Y, Sun Y, Liu L and Li L: miR-19a protects cardiomyocytes from hypoxia/reoxygenation-induced apoptosis via PTEN/PI3K/p-Akt pathway. Biosci Rep. 37:BSR201708992017. View Article : Google Scholar : PubMed/NCBI
4	Wang X, Ha T, Hu Y, Lu C, Liu L, Zhang X, Kao R, Kalbfleisch J, Williams D and Li C: MicroRNA-214 protects against hypoxia/reoxygenation induced cell damage and myocardial ischemia/reperfusion injury via suppression of PTEN and Bim1 expression. Oncotarget. 7:86926–86936. 2016. View Article : Google Scholar : PubMed/NCBI
5	Qiu R, Li W and Liu Y: MicroRNA-204 protects H9C2 cells against hypoxia/reoxygenation-induced injury through regulating SIRT1-mediated autophagy. Biomed Pharmacother. 100:15–19. 2018. View Article : Google Scholar : PubMed/NCBI
6	Kawaguchi M, Takahashi M, Hata T, Kashima Y, Usui F, Morimoto H, Izawa A, Takahashi Y, Masumoto J, Koyama J, et al: Inflammasome activation of cardiac fibroblasts is essential for myocardial ischemia/reperfusion injury. Circulation. 123:594–604. 2011. View Article : Google Scholar : PubMed/NCBI
7	Feng R, Liu J, Wang Z, Zhang J, Cates C, Rousselle T, Meng Q and Li J: The structure-activity relationship of ginsenosides on hypoxia-reoxygenation induced apoptosis of cardiomyocytes. Biochem Biophys Res Commun. 494:556–568. 2017. View Article : Google Scholar : PubMed/NCBI
8	Li Y, Shi X, Li J, Zhang M and Yu B: Knockdown of KLF11 attenuates hypoxia/reoxygenation injury via JAK2/STAT3 signaling in H9c2. Apoptosis. 22:510–518. 2017. View Article : Google Scholar : PubMed/NCBI
9	Tutar Y: miRNA and cancer; computational and experimental approaches. Curr Pharm Biotechnol. 15:4292014. View Article : Google Scholar : PubMed/NCBI
10	Lesizza P, Prosdocimo G, Martinelli V, Sinagra G, Zacchigna S and Giacca M: Single-dose intracardiac injection of Pro-regenerative MicroRNAs improves cardiac function after myocardial infarction. Circ Res. 120:1298–1304. 2017. View Article : Google Scholar : PubMed/NCBI
11	Li YG, Zhang PP, Jiao KL and Zou YZ: Knockdown of microRNA-181 by lentivirus mediated siRNA expression vector decreases the arrhythmogenic effect of skeletal myoblast transplantation in rat with myocardial infarction. Microvasc Res. 78:393–404. 2009. View Article : Google Scholar : PubMed/NCBI
12	Wang J, Bai Y, Li N, Ye W, Zhang M, Greene SB, Tao Y, Chen Y, Wehrens XH and Martin JF: Pitx2-microRNA pathway that delimits sinoatrial node development and inhibits predisposition to atrial fibrillation. Proc Natl Acad Sci USA. 111:9181–6. 2014. View Article : Google Scholar : PubMed/NCBI
13	Rodriguez A, Vigorito E, Clare S, Warren MV, Couttet P, Soond DR, van Dongen S, Grocock RJ, Das PP, Miska EA, et al: Requirement of bic/microRNA-155 for normal immune function. Science. 316:608–611. 2007. View Article : Google Scholar : PubMed/NCBI
14	Elton TS, Selemon H, Elton SM and Parinandi NL: Regulation of the MIR155 host gene in physiological and pathological processes. Gene. 532:1–12. 2013. View Article : Google Scholar : PubMed/NCBI
15	Faraoni I, Antonetti FR, Cardone J and Bonmassar E: miR-155 gene: A typical multifunctional microRNA. Biochim Biophys Acta. 1792:497–505. 2009. View Article : Google Scholar : PubMed/NCBI
16	Nazari-Jahantigh M, Wei Y, Noels H, Akhtar S, Zhou Z, Koenen RR, Heyll K, Gremse F, Kiessling F, Grommes J, et al: MicroRNA-155 promotes atherosclerosis by repressing Bcl6 in macrophages. J Clin Invest. 122:4190–202. 2012. View Article : Google Scholar : PubMed/NCBI
17	Eisenhardt SU, Weiss JB, Smolka C, Maxeiner J, Pankratz F, Bemtgen X, Kustermann M, Thiele JR, Schmidt Y, Bjoern Stark G, et al: MicroRNA-155 aggravates ischemia-reperfusion injury by modulation of inflammatory cell recruitment and the respiratory oxidative burst. Basic Res Cardiol. 110:322015. View Article : Google Scholar : PubMed/NCBI
18	Nederlof R, Eerbeek O, Hollmann MW, Southworth R and Zuurbier CJ: Targeting hexokinase II to mitochondria to modulate energy metabolism and reduce ischaemia-reperfusion injury in heart. Br J Pharmacol. 171:2067–2079. 2014. View Article : Google Scholar : PubMed/NCBI
19	Tan L, Jiang W, Lu A, Cai H and Kong L: miR-155 aggravates liver Ischemia/reperfusion injury by suppressing SOCS1 in mice. Transplant Proc. 50:3831–3839. 2018. View Article : Google Scholar : PubMed/NCBI
20	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. View Article : Google Scholar : PubMed/NCBI
21	Enright AJ, John B, Gaul U, Tuschl T, Sander C and Marks DS: MicroRNA targets in drosophila. Genome Biol. 5:R12003. View Article : Google Scholar : PubMed/NCBI
22	Dweep H and Gretz N: miRWalk2.0: A comprehensive atlas of microRNA-target interactions. Nat Methods. 12:6972015. View Article : Google Scholar : PubMed/NCBI
23	Lewis BP, Burge CB and Bartel DP: Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets. Cell. 120:15–20. 2005. View Article : Google Scholar : PubMed/NCBI
24	Kalia SK, Kalia LV and McLean PJ: Molecular chaperones as rational drug targets for Parkinson's disease therapeutics. CNS Neurol Disord Drug Targets. 9:741–753. 2010. View Article : Google Scholar : PubMed/NCBI
25	Harper SJ and LoGrasso P: Signalling for survival and death in neurones: The role of stress-activated kinases, JNK and p38. Cell Signal. 13:299–310. 2001. View Article : Google Scholar : PubMed/NCBI
26	Lamb JA, Ventura JJ, Hess P, Flavell RA and Davis RJ: JunD mediates survival signaling by the JNK signal transduction pathway. Mol Cell. 11:1479–1489. 2003. View Article : Google Scholar : PubMed/NCBI
27	Chan SH, Hsu KS, Huang CC, Wang LL, Ou CC and Chan JY: NADPH oxidase-derived superoxide anion mediates angiotensin II-induced pressor effect via activation of p38 mitogen-activated protein kinase in the rostral ventrolateral medulla. Circ Res. 97:772–780. 2005. View Article : Google Scholar : PubMed/NCBI
28	Shao Z, Bhattacharya K, Hsich E, Park L, Walters B, Germann U, Wang YM, Kyriakis J, Mohanlal R, Kuida K, et al: c-Jun N-terminal kinases mediate reactivation of Akt and cardiomyocyte survival after hypoxic injury in vitro and in vivo. Circ Res. 98:111–118. 2006. View Article : Google Scholar : PubMed/NCBI
29	Kaelin WG Jr and Ratcliffe PJ: Oxygen sensing by metazoans: The central role of the HIF hydroxylase pathway. Mol Cell. 30:393–402. 2008. View Article : Google Scholar : PubMed/NCBI
30	Semenza GL: HIF-1 inhibitors for cancer therapy: From gene expression to drug discovery. Curr Pharm Des. 15:3839–3843. 2009. View Article : Google Scholar : PubMed/NCBI
31	Epstein AC, Gleadle JM, McNeill LA, Hewitson KS, O'Rourke J, Mole DR, Mukherji M, Metzen E, Wilson MI, Dhanda A, et al: C. elegans EGL-9 and mammalian homologs define a family of dioxygenases that regulate HIF by prolyl hydroxylation. Cell. 107:43–54. 2001. View Article : Google Scholar : PubMed/NCBI
32	Brennan J: Reperfusion injury of cardiac myocytes: Mechanisms, treatment, and implications for advanced practice nursing. AACN Clin Issues. 11:252–260. 2000. View Article : Google Scholar : PubMed/NCBI
33	Hausenloy DJ and Yellon DM: Myocardial ischemia-reperfusion injury: A neglected therapeutic target. J Clin Invest. 123:92–100. 2013. View Article : Google Scholar : PubMed/NCBI
34	Baines CP: How and when do myocytes die during ischemia and reperfusion: The late phase. J Cardiovasc Pharmacol Ther. 16:239–243. 2011. View Article : Google Scholar : PubMed/NCBI
35	Braunwald E: The war against heart failure: The lancet lecture. Lancet. 385:812–824. 2015. View Article : Google Scholar : PubMed/NCBI
36	Zhao ZQ: Oxidative stress-elicited myocardial apoptosis during reperfusion. Curr Opin Pharmacol. 4:159–165. 2004. View Article : Google Scholar : PubMed/NCBI
37	Podsiad A, Standiford TJ, Ballinger MN, Eakin R, Park P, Kunkel SL, Moore BB and Bhan U: MicroRNA-155 regulates host immune response to postviral bacterial pneumonia via IL-23/IL-17 pathway. Am J Physiol Lung Cell Mol Physiol. 310:L465–L475. 2016. View Article : Google Scholar : PubMed/NCBI
38	Bayraktar R and Van Roosbroeck K: miR-155 in cancer drug resistance and as target for miRNA-based therapeutics. Cancer Metastasis Rev. 37:33–44. 2018. View Article : Google Scholar : PubMed/NCBI
39	Gangwar RS, Rajagopalan S, Natarajan R and Deiuliis JA: Noncoding RNAs in cardiovascular disease: Pathological relevance and emerging role as biomarkers and therapeutics. Am J Hypertens. 31:150–165. 2018. View Article : Google Scholar : PubMed/NCBI
40	Zhai A, Qian J, Kao W, Li A, Li Y, He J, Zhang Q, Song W, Fu Y, Wu J, et al: Borna disease virus encoded phosphoprotein inhibits host innate immunity by regulating miR-155. Antiviral Res. 98:66–75. 2013. View Article : Google Scholar : PubMed/NCBI
41	Cao S, Wang Y, Li J, Lv M, Niu H and Tian Y: Tumor-suppressive function of long noncoding RNA MALAT1 in glioma cells by suppressing miR-155 expression and activating FBXW7 function. Am J Cancer Res. 6:2561–2574. 2016.PubMed/NCBI
42	Teramura T and Onodera Y: Stem cell depletion by inflammation-associated miR-155. Aging (Albany NY). 10:17–18. 2018. View Article : Google Scholar : PubMed/NCBI
43	Tili E, Mezache L, Michaille JJ, Amann V, Williams J, Vandiver P, Quinonez M, Fadda P, Mikhail A and Nuovo G: microRNA 155 up regulation in the CNS is strongly correlated to Down's syndrome dementia. Ann Diagn Pathol. 34:103–109. 2018. View Article : Google Scholar : PubMed/NCBI
44	Bruchmann A, Roller C, Walther TV, Schafer G, Lehmusvaara S, Visakorpi T, Klocker H, Cato AC and Maddalo D: Bcl-2 associated athanogene 5 (Bag5) is overexpressed in prostate cancer and inhibits ER-stress induced apoptosis. BMC Cancer. 13:962013. View Article : Google Scholar : PubMed/NCBI
45	Guo K, Li L, Yin G, Zi X and Liu L: Bag5 protects neuronal cells from amyloid β-induced cell death. J Mol Neurosci. 55:815–820. 2015. View Article : Google Scholar : PubMed/NCBI
46	Ying Z, Haiyan G and Haidong G: BAG5 regulates PTEN stability in MCF-7 cell line. BMB Rep. 46:490–494. 2013. View Article : Google Scholar : PubMed/NCBI
47	Yue X, Zhao Y, Huang G, Li J, Zhu J, Feng Z and Hu W: A novel mutant p53 binding partner BAG5 stabilizes mutant p53 and promotes mutant p53 GOFs in tumorigenesis. Cell Discov. 2:160392016. View Article : Google Scholar : PubMed/NCBI
48	Kalia SK, Lee S, Smith PD, Liu L, Crocker SJ, Thorarinsdottir TE, Glover JR, Fon EA, Park DS and Lozano AM: BAG5 inhibits parkin and enhances dopaminergic neuron degeneration. Neuron. 44:931–945. 2004. View Article : Google Scholar : PubMed/NCBI
49	Bi L, Yang Q, Yuan J, Miao Q, Duan L, Li F and Wang S: MicroRNA-127-3p acts as a tumor suppressor in epithelial ovarian cancer by regulating the BAG5 gene. Oncol Rep. 36:2563–2570. 2016. View Article : Google Scholar : PubMed/NCBI
50	Bourke LT, McDonnell T, McCormick J, Pericleous C, Ripoll VM, Giles I, Rahman A, Stephanou A and Ioannou Y: Antiphospholipid antibodies enhance rat neonatal cardiomyocyte apoptosis in an in vitro hypoxia/reoxygenation injury model via p38 MAPK. Cell Death Dis. 8:e25492017. View Article : Google Scholar : PubMed/NCBI
51	Li Q, Xiang Y, Chen Y, Tang Y and Zhang Y: Ginsenoside Rg1 protects cardiomyocytes against hypoxia/reoxygenation injury via activation of Nrf2/HO-1 signaling and inhibition of JNK. Cell Physiol Biochem. 44:21–37. 2017. View Article : Google Scholar : PubMed/NCBI