Ischemic postconditioning attenuates the inflammatory response in ischemia/reperfusion myocardium by upregulating miR‑499 and inhibiting TLR2 activation

Zhang,Xin‑Yue; Huang,Zheng; Li,Qing‑Jie; Zhong,Guo‑Qiang; Meng,Jian‑Jun; Wang,Dong‑Xiao; Tu,Rong‑Hui

doi:10.3892/mmr.2020.11104

Ischemic postconditioning attenuates the inflammatory response in ischemia/reperfusion myocardium by upregulating miR‑499 and inhibiting TLR2 activation

Authors:
- Xin‑Yue Zhang
- Zheng Huang
- Qing‑Jie Li
- Guo‑Qiang Zhong
- Jian‑Jun Meng
- Dong‑Xiao Wang
- Rong‑Hui Tu
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Affiliations: Department of Cardiology, First Affiliated Hospital, Guangxi Medical University, Nanning, Guangxi 530021, P.R. China, Department of Cardiology, Second Affiliated Hospital, Guangxi Medical University, Nanning, Guangxi 530021, P.R. China, Department of Geriatric Health Care Center, First Affiliated Hospital, Guangxi Medical University, Nanning, Guangxi 530021, P.R. China, Guangxi Key Laboratory of Precision Medicine in Cardio‑Cerebrovascular Diseases Control and Prevention, Nanning, Guangxi 530021, P.R. China
Published online on: April 30, 2020 https://doi.org/10.3892/mmr.2020.11104
Pages: 209-218
Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Toll-like receptor 2 (TLR2)-mediated myocardial inflammation serves an important role in promoting myocardial ischemic/reperfusion (I/R) injury. Previous studies have shown that miR‑499 is critical for cardioprotection after ischemic postconditioning (IPostC). Therefore, the present study evaluated the protective effect of IPostC on the myocardium by inhibiting TLR2, and also assessed the involvement of microRNA (miR)‑499. Rat hearts were subjected to 30 min of ischemia and 2 h of reperfusion. The IPostC was 3 cycles of 30 sec of reperfusion and 30 sec of re‑occlusion prior to reperfusion. In total, 90 rats were randomly divided into six groups (n=15 per group): Sham; I/R; IPostC; miR‑499 negative control adeno‑associated virus (AAV) vectors + IPostC; miR‑499 inhibitor AAV vectors + IPostC; and miR‑499 mimic AAV vectors + IPostC. It was identified that IPostC significantly decreased the I/R‑induced cardiomyocyte apoptotic index (29.4±2.03% in IPostC vs. 42.64±2.27% in I/R; P<0.05) and myocardial infarct size (48.53±2.49% in IPostC vs. 66.52±3.1% in I/R; P<0.05). Moreover, these beneficial effects were accompanied by increased miR‑499 expression levels (as demonstrated by reverse transcription‑quantitative PCR) in the myocardial tissue and decreased TLR2, protein kinase C (PKC), interleukin (IL)‑1β and IL‑6 expression levels (as demonstrated by western blotting and ELISA) in the myocardium and serum. The results indicated that IPostC + miR‑499 mimics significantly inhibited inflammation and the PKC signaling pathway and enhanced the anti‑inflammatory and anti‑apoptotic effects of IPostC. However, IPostC + miR‑499 inhibitors had the opposite effect. Therefore, it was speculated that IPostC may have a miR‑499‑dependent cardioprotective effect. The present results suggested that miR‑499 may be involved in IPostC‑mediated ischemic cardioprotection, which may occur via local and systemic TLR2 inhibition, subsequent inhibition of the PKC signaling pathway and a decrease in inflammatory cytokine release, including IL‑1β and IL‑6. Moreover, these effects will ultimately lead to a decrease in the myocardial apoptotic index and myocardial infarct size via the induction of the anti‑apoptotic protein Bcl‑2, and inhibition of the pro‑apoptotic protein Bax in myocardium.

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1	Turer AT and Hill JA: Pathogenesis of myocardial ischemia-reperfusion injury and rationale for therapy. Am J Cardiol. 106:360–368. 2010. View Article : Google Scholar : PubMed/NCBI
2	Hausenloy DJ, Botker HE, Engstrom T, Erlinge D, Heusch G, Ibanez B, Kloner RA, Ovize M, Yellon DM and Garcia-Dorado D: Targeting reperfusion injury in patients with ST-segment elevation myocardial infarction: Trials and tribulations. Eur Heart J. 38:935–941. 2017.PubMed/NCBI
3	Yang Y, Yang J, Liu XW, Ding JW, Li S, Guo X, Yang CJ, Fan ZX, Wang HB, Li Q, et al: Down-Regulation of miR-327 alleviates ischemia/reperfusion-induced myocardial damage by targeting RP105. Cell Physiol Biochem. 49:1049–1063. 2018. View Article : Google Scholar : PubMed/NCBI
4	Pope MR and Fleming SD: TLR2 modulates antibodies required for intestinal ischemia/reperfusion-induced damage and inflammation. J Immunol. 194:1190–1198. 2015. View Article : Google Scholar : PubMed/NCBI
5	Arslan F, Keogh B, Mcguirk P and Parker AE: TLR2 and TLR4 in ischemia reperfusion injury. Mediators Inflamm. 2010:7042022010. View Article : Google Scholar : PubMed/NCBI
6	Chen E, Bakr MM, Firth N and Love RM: Inflammatory cell expression of Toll-like receptor-2 (TLR2) within refractory periapical granuloma. F1000Res. 7:18192018. View Article : Google Scholar : PubMed/NCBI
7	Ward JR, Wilson HL, Francis SE, Crossman DC and Sabroe I: Translational mini-review series on immunology of vascular disease: Inflammation, infections and Toll-like receptors in cardiovascular disease. Clin Exp Immunol. 156:386–394. 2009. View Article : Google Scholar : PubMed/NCBI
8	Ha T, Hu Y, Liu L, Lu C, McMullen JR, Kelley J, Kao RL, Williams DL, Gao X and Li C: TLR2 ligands induce cardioprotection against ischaemia/reperfusion injury through a PI3K/Akt-dependent mechanism. Cardiovasc Res. 87:694–703. 2010. View Article : Google Scholar : PubMed/NCBI
9	Arslan F, Houtgraaf JH, Keogh B, Kazemi K, de Jong R, McCormack WJ, O'Neill LA, McGuirk P, Timmers L, Smeets MB, et al: Treatment with OPN-305, a humanized anti-toll-like receptor-2 antibody, reduces myocardial ischemia/reperfusion injury in pigs. Circ Cardiovasc Interv. 5:279–287. 2012. View Article : Google Scholar : PubMed/NCBI
10	Skyschally A, van Caster P, Iliodromitis EK, Schulz R, Kremastinos DT and Heusch G: Ischemic postconditioning: Experimental models and protocol algorithms. Basic Res Cardiol. 104:469–483. 2009. View Article : Google Scholar : PubMed/NCBI
11	Staat P, Rioufol G, Piot C, Cottin Y, Cung TT, L'Huillier I, Aupetit JF, Bonnefoy E, Finet G, André-Fouët X and Ovize M: Postconditioning the human heart. Circulation. 112:2143–2148. 2005. View Article : Google Scholar : PubMed/NCBI
12	Zhang DW, Zhang L, Liu JG, Wang CL, Shi DZ and Chen KJ: Mechanisms of Chinese herbs combined with ischemic postconditioning in protecting myocardium of rats from ischemia-reperfusion injury. Zhong Xi Yi Jie He Xue Bao. 8:465–471. 2010.(In Chinese). View Article : Google Scholar : PubMed/NCBI
13	Wang Y, Ge P, Yang L, Wu C, Zha H, Luo T and Zhu Y: Protection of ischemic post conditioning against transient focal ischemia-induced brain damage is associated with inhibition of neuroinflammation via modulation of TLR2 and TLR4 pathways. J Neuroinflammation. 11:152014. View Article : Google Scholar : PubMed/NCBI
14	Zhong GQ, Tu RH, Zeng ZY, Li QJ, He Y, Li S, He Y and Xiao F: Novel functional role of heat shock protein 90 in protein kinase C-mediated ischemic postconditioning. J Surg Res. 189:198–206. 2014. View Article : Google Scholar : PubMed/NCBI
15	Sheedy FJ and O'Neill LA: Adding fuel to fire: MicroRNAs as a new class of mediators of inflammation. Ann Rheum Dis. 67 (Suppl 3):iii50–iii55. 2008. View Article : Google Scholar : PubMed/NCBI
16	Wu CJ and Lu LF: MicroRNA in immune regulation. Curr Top Microbiol Immunol. 410:249–267. 2017.PubMed/NCBI
17	Wojciechowska A, Braniewska A and Kozar-Kamińska K: MicroRNA in cardiovascular biology and disease. Adv Clin Exp Med. 26:865–874. 2017. View Article : Google Scholar : PubMed/NCBI
18	Hajibabaie F, Kouhpayeh S, Mirian M, Rahimmanesh I, Boshtam M, Sadeghian L, Gheibi A, Khanahmad H and Shariati L: MicroRNAs as the actors in the atherosclerosis scenario. J Physiol Biochem. 76:1–12. 2020. View Article : Google Scholar : PubMed/NCBI
19	Li Y, Lu J, Bao X, Wang X, Wu J, Li X and Hong W: MiR-499-5p protects cardiomyocytes against ischaemic injury via anti-apoptosis by targeting PDCD4. Oncotarget. 7:35607–35617. 2016. View Article : Google Scholar : PubMed/NCBI
20	Jia Z, Wang J, Shi Q, Liu S, Wang W, Tian Y, Lu Q, Chen P, Ma K and Zhou C: SOX6 and PDCD4 enhance cardiomyocyte apoptosis through LPS-induced miR-499 inhibition. Apoptosis. 21:174–183. 2016. View Article : Google Scholar : PubMed/NCBI
21	Zhu J, Yao K, Wang Q, Guo J, Shi H, Ma L, Liu H, Gao W, Zou Y and Ge J: Ischemic postconditioning-regulated miR-499 protects the rat heart against ischemia/reperfusion injury by inhibiting apoptosis through PDCD4. Cell Physiol Biochem. 39:2364–2380. 2016. View Article : Google Scholar : PubMed/NCBI
22	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
23	Sheibani M, Faghir-Ghanesefat H, Dehpour S, Keshavarz-Bahaghighat H, Sepand MR, Ghahremani MH, Azizi Y, Rahimi N and Dehpour AR: Sumatriptan protects against myocardial ischaemia-reperfusion injury by inhibition of inflammation in rat model. Inflammopharmacology. 27:1071–1080. 2019. View Article : Google Scholar : PubMed/NCBI
24	Sakata Y, Dong JW, Vallejo JG, Huang CH, Baker JS, Tracey KJ, Tacheuchi O, Akira S and Mann DL: Toll-like receptor 2 modulates left ventricular function following ischemia-reperfusion injury. Am J Physiol Heart Circ Physiol. 292:H503–H509. 2007. View Article : Google Scholar : PubMed/NCBI
25	Selejan S, Poss J, Walter F, Hohl M, Kaiser R, Kazakov A, Böhm M and Link A: Ischaemia-induced up-regulation of Toll-like receptor 2 in circulating monocytes in cardiogenic shock. Eur Heart J. 33:1085–1094. 2012. View Article : Google Scholar : PubMed/NCBI
26	Arslan F, Smeets MB, O'Neill LA, Keogh B, McGuirk P, Timmers L, Tersteeg C, Hoefer IE, Doevendans PA, Pasterkamp G and de Kleijn DP: Myocardial ischemia/reperfusion injury is mediated by leukocytic toll-like receptor-2 and reduced by systemic administration of a novel anti-toll-like receptor-2 antibody. Circulation. 121:80–90. 2010. View Article : Google Scholar : PubMed/NCBI
27	Mersmann J, Berkels R, Zacharowski P, Tran N, Koch A, Iekushi K, Dimmeler S, Granja TF, Boehm O, Claycomb WC and Zacharowski K: Preconditioning by toll-like receptor 2 agonist Pam3CSK4 reduces CXCL1-dependent leukocyte recruitment in murine myocardial ischemia/reperfusion injury. Crit Care Med. 38:903–909. 2010. View Article : Google Scholar : PubMed/NCBI
28	Chatterjee PK: Cardiac preconditioning by specific ligands of Toll-like receptors: Is it wither or whither? Crit Care Med. 38:1003–1004. 2010. View Article : Google Scholar : PubMed/NCBI
29	Dong JW, Vallejo JG, Tzeng HP, Thomas JA and Mann DL: Innate immunity mediates myocardial preconditioning through Toll-like receptor 2 and TIRAP-dependent signaling pathways. Am J Physiol Heart Circ Physiol. 298:H1079–H1087. 2010. View Article : Google Scholar : PubMed/NCBI
30	Vilahur G and Badimon L: Ischemia/reperfusion activates myocardial innate immune response: The key role of the toll-like receptor. Front Physiol. 5:4962014. View Article : Google Scholar : PubMed/NCBI
31	Jin C, Cleveland JC, Ao L, Li J, Zeng Q, Fullerton DA and Meng X: Human myocardium releases heat shock protein 27 (HSP27) after global ischemia: The proinflammatory effect of extracellular HSP27 through toll-like receptor (TLR)-2 and TLR4. Mol Med. 20:280–289. 2014. View Article : Google Scholar : PubMed/NCBI
32	Jong WM, Ten Cate H, Linnenbank AC, de Boer OJ, Reitsma PH, de Winter RJ and Zuurbier CJ: Reduced acute myocardial ischemia-reperfusion injury in IL-6-deficient mice employing a closed-chest model. Inflamm Res. 65:489–499. 2016. View Article : Google Scholar : PubMed/NCBI
33	De Jesus NM, Wang L, Lai J, Rigor RR, Francis Stuart SD, Bers DM, Lindsey ML and Ripplinger CM: Antiarrhythmic effects of interleukin 1 inhibition after myocardial infarction. Heart Rhythm. 14:727–736. 2017. View Article : Google Scholar : PubMed/NCBI
34	Kharbanda RK, Mortensen UM, White PA, Kristiansen SB, Schmidt MR, Hoschtitzky JA, Vogel M, Sorensen K, Redington AN and MacAllister R: Transient limb ischemia induces remote ischemic preconditioning in vivo. Circulation. 106:2881–2883. 2002. View Article : Google Scholar : PubMed/NCBI
35	Przyklenk K, Bauer B, Ovize M, Kloner RA and Whittaker P: Regional ischemic ‘preconditioning’ protects remote virgin myocardium from subsequent sustained coronary occlusion. Circulation. 87:893–899. 1993. View Article : Google Scholar : PubMed/NCBI
36	Wang S, Zhang F, Zhao G, Cheng Y, Wu T, Wu B and Zhang YE: Mitochondrial PKC-εdeficiency promotes I/R-mediated myocardial injury via GSK3β-dependent mitochondrial permeability transition pore opening. J Cell Mol Med. 21:2009–2021. 2017. View Article : Google Scholar : PubMed/NCBI
37	Zheng H, Liu J, Liu C, Lu F, Zhao Y, Jin Z, Ren H, Leng X, Jia J, Hu G, et al: Calcium-sensing receptor activating phosphorylation of PKCδ translocation on mitochondria to induce cardiomyocyte apoptosis during ischemia/reperfusion. Mol Cell Biochem. 358:335–343. 2011. View Article : Google Scholar : PubMed/NCBI
38	Cario E, Gerken G and Podolsky DK: Toll-like receptor 2 enhances ZO-1-associated intestinal epithelial barrier integrity via protein kinase C. Gastroenterology. 127:224–238. 2004. View Article : Google Scholar : PubMed/NCBI
39	Park DW, Lee HK, Lyu JH, Chin H, Kang SW, Kim YJ, Bae YS and Baek SH: TLR2 stimulates ABCA1 expression via PKC-η and PLD2 pathway. Biochem Biophys Res Commun. 430:933–937. 2013. View Article : Google Scholar : PubMed/NCBI
40	Jiang SY, Wei CC, Shang TT, Lian Q, Wu CX and Deng JY: High glucose induces inflammatory cytokine through protein kinase C-induced toll-like receptor 2 pathway in gingival fibroblasts. Biochem Biophys Res Commun. 427:666–670. 2012. View Article : Google Scholar : PubMed/NCBI
41	Xin Y, Yang C and Han Z: Circulating miR-499 as a potential biomarker for acute myocardial infarction. Ann Transl Med. 4:1352016. View Article : Google Scholar : PubMed/NCBI
42	Liu X, Fan Z, Zhao T, Cao W, Zhang L, Li H, Xie Q, Tian Y and Wang B: Plasma miR-1, miR-208, miR-499 as potential predictive biomarkers for acute myocardial infarction: An independent study of Han population. Exp Gerontol. 72:230–238. 2015. View Article : Google Scholar : PubMed/NCBI
43	Qin H, Chen GX, Liang MY, Rong J, Yao JP, Liu H and Wu ZK: The altered expression profile of microRNAs in cardiopulmonary bypass canine models and the effects of mir-499 on myocardial ischemic reperfusion injury. J Transl Med. 11:1542013. View Article : Google Scholar : PubMed/NCBI