miR‑217 and miR‑543 downregulation mitigates inflammatory response and myocardial injury in children with viral myocarditis by regulating the SIRT1/AMPK/NF‑κB signaling pathway

Xia,Kun; Zhang,Yong; Sun,Dongming

doi:10.3892/ijmm.2019.4442

miR‑217 and miR‑543 downregulation mitigates inflammatory response and myocardial injury in children with viral myocarditis by regulating the SIRT1/AMPK/NF‑κB signaling pathway

Authors:
- Kun Xia
- Yong Zhang
- Dongming Sun
View Affiliations

Affiliations: Department of Cardiovascular Medicine, Wuhan Children's Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430019, P.R China
Published online on: December 27, 2019 https://doi.org/10.3892/ijmm.2019.4442
Pages: 634-646

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

The aim of the present study was to investigate the expression levels and roles of microRNA (miR)‑217 and miR‑543 in viral myocarditis, and to examine their underlying mechanisms. Coxsackievirus B3 (CVB3) was used to establish in vivo and in vitro models of viral myocarditis. The levels of miR‑217 and miR‑543 were detected using reverse transcription‑quantitative PCR. The association between miR‑217 and miR‑543 and sirtuin‑1 (SIRT1) was predicted and confirmed by TargetScan and dual‑luciferase reporter assay. Cell viability was detected using Cell Counting Kit‑8 assay, and cell apoptosis was measured by analyzing the expression levels of Bcl‑2 and Bax, and by flow cytometry. In addition, the synthesis of various pro‑inflammatory factors was determined by ELISA. In addition, superoxide dismutase (SOD) activity and malondialdehyde (MDA) levels were measured in cardiomyocytes following transfection and CVB infection. miR‑217 and miR‑543 were found to be highly expressed in the peripheral blood of pediatric patients with viral myocarditis, in the peripheral blood and myocardial tissues of viral myocarditis mice and in CVB3‑infected cardiomyocytes. SIRT1 was found to be a target of both miR‑217 and miR‑543, and SIRT1 expression level was downregulated in viral myocarditis. Further analysis indicated that the reduced cell viability, increased cell apoptosis, enhanced synthesis of inflammatory factors, increased MDA content and decreased SOD activity associated with myocarditis were significantly reversed after inhibition of miR‑217 or miR‑543. Importantly, the present results showed that all the effects of miR‑217 and miR‑543 inhibition on cardiomyocytes were significantly suppressed following SIRT1 knockdown. Collectively, the present data indicated that miR‑217 and miR‑543 were significantly upregulated in viral myocarditis, and downregulation of miR‑217 and miR‑543 attenuated CVB3 infection‑induced cardiomyocyte injury by targeting SIRT1. miR‑217 and miR‑543 may be potential therapeutic targets for developing novel viral myocarditis treatments in the future.

View Figures

View References

1	Yajima T: Viral myocarditis: Potential defense mechanisms within the cardiomyocyte against virus infection. Future Microbiol. 6:551–566. 2011. View Article : Google Scholar : PubMed/NCBI
2	Gaaloul I, Riabi S, Harrath R, Evans M, Salem NH, Mlayeh S, Huber S and Aouni M: Sudden unexpected death related to enterovirus myocarditis: Histopathology, immunohistochemistry and molecular pathology diagnosis at post-mortem. BMC Infect Dis. 12:2122012. View Article : Google Scholar : PubMed/NCBI
3	Wang T, Zhang J, Xiao A, Liu W, Shang Y and An J: Melittin ameliorates CVB3-induced myocarditis via activation of the HDAC2-mediated GSK-3β/Nrf2/ARE signaling pathway. Biochem Biophys Res Commun. 480:126–131. 2016. View Article : Google Scholar : PubMed/NCBI
4	Márquez-González H, López-Gallegos D and González- Espi nosa AM: Effect of immune therapy in the prognosis of viral myocarditis in pediatric patients. Rev Med Inst Mex Seguro Soc. 54(Suppl 3): S296–S301. 2016.In Spanish.
5	Yu M, Long Q, Li HH, Liang W, Liao YH, Yuan J and Cheng X: IL-9 inhibits viral replication in coxsackievirus B3-induced myocarditis. Front Immunol. 7:4092016. View Article : Google Scholar : PubMed/NCBI
6	Wang C, Dong C and Xiong S: IL-33 enhances macrophage M2 polarization and protects mice from CVB3-induced viral myocarditis. J Mol Cell Cardiol. 103:22–30. 2017. View Article : Google Scholar : PubMed/NCBI
7	Pollack A, Kontorovich AR, Fuster V and Dec GW: Viral myocarditis-diagnosis, treatment options, and current controversies. Nat Rev Cardiol. 12:670–680. 2015. View Article : Google Scholar : PubMed/NCBI
8	Rienks M, Papageorgiou A, Wouters K, Verhesen W, Leeuwen RV, Carai P, Summer G, Westermann D and Heymans SA: novel 72-kDa leukocyte-derived osteoglycin enhances the activation of toll-like receptor 4 and exacerbates cardiac inflammation during viral myocarditis. Cell Mol Life Sci. 74:1511–1525. 2017. View Article : Google Scholar
9	Yue-Chun L, Guang-Yi C, Li-Sha G, Chao X, Xinqiao T, Cong L, Xiao-Ya D and Xiangjun Y: The protective effects of ivabradine in preventing progression from viral myocarditis to dilated cardiomyopathy. Front Pharmacol. 7:4082016. View Article : Google Scholar : PubMed/NCBI
10	Bartel DP: MicroRNAs: Genomics, biogenesis, mechanism, and function. Cell. 116:281–297. 2004. View Article : Google Scholar : PubMed/NCBI
11	Hammond SM: An overview of microRNAs. Adv Drug Deliv Rev. 87:3–14. 2015. View Article : Google Scholar : PubMed/NCBI
12	Ghildiyal M and Zamore PD: Small silencing RNAs: An expanding universe. Nat Rev Genet. 10:94–108. 2009. View Article : Google Scholar : PubMed/NCBI
13	Soifer HS, Rossi JJ and Saetrom P: MicroRNAs in disease and potential therapeutic applications. Mol Ther. 15:2070–2079. 2007. View Article : Google Scholar : PubMed/NCBI
14	Krol J, Loedige I and Filipowicz W: The widespread regulation of microRNA biogenesis, function and decay. Nat Rev Genet. 11:597–610. 2010. View Article : Google Scholar : PubMed/NCBI
15	O'Connell RM, Rao DS, Chaudhuri AA and Baltimore D: Physiological and pathological roles for microRNAs in the immune system. Nat Rev Immunol. 10:111–122. 2010. View Article : Google Scholar : PubMed/NCBI
16	Bardooli F, McAlindon E, Littlejohns B, Suleiman MS, Bucciarelli-Ducci C and Baumbach A: TCT-184 Early changes in circulating miRNA 133a are indicative of cardiac remodelling after 3 months in patients presenting with acute ST elevation myocardial infarction. J Am Coll Cardiol. 68:B75–B76. 2016. View Article : Google Scholar
17	Wang Y, Ouyang M, Wang Q and Jian Z: MicroRNA-142-3p inhibits hypoxia/reoxygenation-induced apoptosis and fibrosis of cardiomyocytes by targeting high mobility group box 1. Int J Mol Med. 38:1377–1386. 2016. View Article : Google Scholar : PubMed/NCBI
18	Singh GB, Raut SK, Khanna S, Kumar A, Sharma S, Prasad R and Khullar M: MicroRNA-200c modulates DUSP-1 expression in diabetes-induced cardiac hypertrophy. Mol Cell Biochem. 424:1–11. 2017. View Article : Google Scholar
19	Corsten MF, Papageorgiou A, Verhesen W, Carai P, Lindow M, Obad S, Summer G, Coort SL, Hazebroek M, van Leeuwen R, et al: MicroRNA profiling identifies microRNA-155 as an adverse mediator of cardiac injury and dysfunction during acute viral myocarditis. Circ Res. 111:415–425. 2012. View Article : Google Scholar : PubMed/NCBI
20	Corsten MF, Dennert R, Jochems S, Kuznetsova T, Devaux Y, Hofstra L, Wagner DR, Staessen JA, Heymans S and Schroen B: Circulating MicroRNA-208b and MicroRNA-499 reflect myocardial damage in cardiovascular disease. Circ Cardiovasc Genet. 3:499–506. 2010. View Article : Google Scholar : PubMed/NCBI
21	Bao JL and Lin L: MiR-155 and miR-148a reduce cardiac injury by inhibiting NF-κB pathway during acute viral myocarditis. Eur Rev Med Pharmacol Sci. 18:2349–2356. 2014.PubMed/NCBI
22	Nogueiras R, Habegger KM, Chaudhary N, Finan B, Banks AS, Dietrich MO, Horvath TL, Sinclair DA, Pfluger PT and Tschöp MH: Sirtuin 1 and sirtuin 3: Physiological modulators of metabolism. Physiol Rev. 92:1479–1514. 2012. View Article : Google Scholar : PubMed/NCBI
23	da Cunha MSB and Arruda SF: Tucum-do-Cerrado (Bactris setosa Mart.) may promote anti-aging effect by upregulating SIRT1-Nrf2 pathway and attenuating oxidative stress and inflammation. Nutrients. 9:pii: E1243. 2017. View Article : Google Scholar : PubMed/NCBI
24	Rada P, Pardo V, Mobasher MA, García-Martínez I, Ruiz L, González-Rodríguez Á, Sanchez-Ramos C, Muntané J, Alemany S, James LP, et al: SIRT1 controls acetaminophen hepatotoxicity by modulating inflammation and oxidative stress. Antioxid Redox Signal. 28:1187–1208. 2018. View Article : Google Scholar
25	Chan SH, Hung CH, Shih JY, Chu PM, Cheng YH, Lin HC and Tsai KL: SIRT1 inhibition causes oxidative stress and inflammation in patients with coronary artery disease. Redox Biol. 13:301–309. 2017. View Article : Google Scholar : PubMed/NCBI
26	Cheng YY, Kao CL, Ma HI, Hung CH, Wang CT, Liu DH, Chen PY and Tsai KL: SIRT1-related inhibition of pro-inflammatory responses and oxidative stress are involved in the mechanism of nonspecific low back pain relief after exercise through modulation of Toll-like receptor 4. J Biochem. 158:299–308. 2015. View Article : Google Scholar : PubMed/NCBI
27	Nguyen LT, Mak CH, Chen H, Zaky AA, Wong MG, Pollock CA and Saad S: SIRT1 attenuates kidney disorders in male offspring due to maternal high-fat diet. Nutrient. 11:pii: E146. 2019. View Article : Google Scholar
28	Hwang JW, Yao H, Caito S, Sundar IK and Rahman I: Redox regulation of SIRT1 in inflammation and cellular senescence. Free Radic Biol Med. 61:95–110. 2013. View Article : Google Scholar : PubMed/NCBI
29	Guo R, Liu W, Liu B, Zhang B, Li W and Xu Y: SIRT1 suppresses cardiomyocyte apoptosis in diabetic cardiomyopathy: An insight into endoplasmic reticulum stress response mechanism. Int J Cardiol. 191:36–45. 2015. View Article : Google Scholar : PubMed/NCBI
30	Mao Q, Liang X, Wu Y and Lu Y: Resveratrol attenuates cardiomyocyte apoptosis in rats induced by coronary micro-embolization through SIRT1-mediated deacetylation of p53. J Cardiovasc Pharmacol Ther. 24:551–558. 2019. View Article : Google Scholar : PubMed/NCBI
31	Zhang WX, He BM, Wu Y, Qiao JF and Peng ZY: Melatonin protects against sepsis-induced cardiac dysfunction by regulating apoptosis and autophagy via activation of SIRT1 in mice. Life Sci. 217:8–15. 2019. View Article : Google Scholar
32	Ben Salem I, Boussabbeh M, Pires Da Silva J, Guilbert A, Bacha H, Abid-Essefi S and Lemaire C: SIRT1 protects cardiac cells against apoptosis induced by zearalenone or its metabolites α- and β-zearalenol through an autophagy-dependent pathway. Toxicol Appl Pharmacol. 314:82–90. 2017. View Article : Google Scholar
33	Li J, Dong G, Wang B, Gao W and Yang Q: miR-543 promotes gastric cancer cell proliferation by targeting SIRT1. Biochem Biophys Res Commun. 469:15–21. 2016. View Article : Google Scholar
34	Hu X, Chi L, Zhang W, Bai T, Zhao W, Feng Z and Tian H: Down-regulation of the miR-543 alleviates insulin resistance through targeting the SIRT1. Biochem Biophys Res Commun. 468:781–787. 2015. View Article : Google Scholar : PubMed/NCBI
35	Yin H, Liang X, Jogasuria A, Davidson NO and You M: miR-217 regulates ethanol-induced hepatic inflammation by disrupting sirtuin 1-lipin-1 signaling. Am J Pathol. 185:1286–1296. 2015. View Article : Google Scholar : PubMed/NCBI
36	Bayne K: Revised guide for the care and use of laboratory animals available American physiological society. Physiologist 39. 199:208–211. 1996.
37	Xin L, Ma X, Xiao Z, Yao H and Liu Z: Coxsackievirus B3 induces autophagy in HeLa cells via the AMPK/MEK/ERK and Ras/Raf/MEK/ERK signaling pathways. Infect Genet Evol. 36:46–54. 2015. View Article : Google Scholar : PubMed/NCBI
38	Qi L, Xin Q and Wenjun J: Inhibition of iNOS protects cardiomyocytes against coxsackievirus B3-induced cell injury by suppressing autophagy. Biomed Pharmacother. 91:673–679. 2017. View Article : Google Scholar : PubMed/NCBI
39	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
40	Tsikas D: Assessment of lipid peroxidation by measuring malondialdehyde (MDA) and relatives in biological samples: Analytical and biological challenges. Anal Biochem. 524:13–30. 2017. View Article : Google Scholar
41	Wu JQ, Kosten TR and Zhang XY: Free radicals, antioxidant defense systems, and schizophrenia. Prog Neuropsychopharmacol Biol Psychiatry. 46:200–206. 2013. View Article : Google Scholar : PubMed/NCBI
42	Casadonte JR, Mazwi ML, Gambetta KE, Palac HL, McBride ME, Eltayeb OM, Monge MC, Backer CL and Costello JM: Risk factors for cardiac arrest or mechanical circulatory support in children with fulminant myocarditis. Pediatr Cardiol. 38:128–134. 2017. View Article : Google Scholar
43	Simpson KE, Storch GA, Lee CK, Ward KE, Danon S, Simon CM, Delaney JW, Tong A and Canter CE: High frequency of detection by PCR of viral nucleic acid in the blood of infants presenting with clinical myocarditis. Pediatr Cardiol. 37:399–404. 2016. View Article : Google Scholar :
44	Tse G, Yeo JM, Chan YW, Lai ET and Yan BP: What is the arrhythmic substrate in viral myocarditis? Insights from clinical and animal studies. Front Physiol. 7:3082016. View Article : Google Scholar : PubMed/NCBI
45	Tian Y, Ma J, Wang W, Zhang L, Xu J, Wang K and Li D: Resveratrol supplement inhibited the NF-κB inflammation pathway through activating AMPKα-SIRT1 pathway in mice with fatty liver. Mol Cell Biochem. 422:75–84. 2016. View Article : Google Scholar : PubMed/NCBI