MiR-103 regulates the angiogenesis of ischemic stroke rats by targeting vascular endothelial growth factor (VEGF)

Document Type : Original Article

Authors

1 Department of Neurology, Affiliated Hospital of Hebei University, Baoding 071000, Hebei Province, China

2 Department of Neurology, the Traditional Chinese Medicine Hospital of Yixian, Baoding 074200, Hebei Province, China

3 Laboratory Animal Center of Hebei University, Hebei University, Baoding 071000, Hebei Province, China

Abstract

Objective(s): To investigate the effect of miR-103 on the angiogenesis of ischemic stroke rats via targeting vascular endothelial growth factor (VEGF) at the molecular level.
Materials and Methods: Rat models had received the middle cerebral artery occlusion (MCAO) or sham operation before grouping, and cell models of oxygen-glucose deprivation (OGD) were performed. FITC-dextran, matrigel, and Trans-well assays were used to evaluate the vascular density, tube formation, and cell migration respectively. The expression levels of miR-103 and VEGF were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting. Dual-luciferase assay was used for analyzing the targeting relationship between miR-103 and VEGF.
Results: We found the reduced miR-103 in rats after MCAO. Down-regulating miR-103 with the miR-103 inhibitor enhanced VEGF, ameliorated the neurological scores, decreased infarct volume, and increased vascular density in rats after MCAO. Besides, in OGD human umbilical vein endothelial cells (HUVECs), inhibition of miR-103 could promote the increase of tube length and the migration of cells. Additionally, we found that miR-103 could directly target VEGF and thereby lead to the down-expression of VEGF. Meanwhile, si-VEGF could reverse the effect of miR-103 inhibitor on angiogenesis in rats subjected to MCAO.
Conclusion: Inhibition of miR-103 could promote ischemic stroke angiogenesis and reduce infarction volume via enhancing VEGF, which provides a new target for the clinical treatment of ischemic stroke.

Keywords

Main Subjects


1. Sun Y, Gui H, Li Q, Luo ZM, Zheng MJ, Duan JL, et al. MicroRNA-124 protects neurons against apoptosis in cerebral ischemic stroke. CNS Neurosci Ther 2013; 19: 813-819.
2. Wei N, Xiao L, Xue R, Zhang D, Zhou J, Ren H, et al. MicroRNA-9 Mediates the Cell Apoptosis by Targeting Bcl2l11 in Ischemic Stroke. Mol Neurobiol 2016; 53: 6809-6817.
3. Zhao WJ, Zhang HF, Su JY. Downregulation of microRNA-195 promotes angiogenesis induced by cerebral infarction via targeting VEGFA. Mol Med Rep 2017; 16: 5434-5440.
4. Truelsen T, Bonita R. Epidemiological transition of stroke in China? Stroke 2008; 39: 1653-1654.
5. Navarro-Sobrino M, Rosell A, Hernandez-Guillamon M, Penalba A, Boada C, Domingues-Montanari S, et al. A large screening of angiogenesis biomarkers and their association with neurological outcome after ischemic stroke. Atherosclerosis 2011; 216: 205-211.
6. Chen J, Chopp M. Neurorestorative treatment of stroke: cell and pharmacological approaches. NeuroRx 2006; 3: 466-473.
7. Manoonkitiwongsa PS, Jackson-Friedman C, McMillan PJ, Schultz RL, Lyden PD. Angiogenesis after stroke is correlated with increased numbers of macrophages: the clean-up hypothesis. J Cereb Blood Flow Metab 2001; 21: 1223-1231.
8. Wei L, Keogh CL, Whitaker VR, Theus MH, Yu SP. Angiogenesis and stem cell transplantation as potential treatments of cerebral ischemic stroke. Pathophysiology 2005; 12: 47-62.
9. Prokopi M, Kousparou CA, Epenetos AA. The Secret Role of microRNAs in Cancer Stem Cell Development and Potential Therapy: A Notch-Pathway Approach. Front Oncol 2014; 4: 389.
10. Caporali A, Emanueli C. MicroRNA regulation in angiogenesis. Vascul Pharmacol 2011; 55: 79-86.
11. Lou YL, Guo F, Liu F, Gao FL, Zhang PQ, Niu X, et al. miR-210 activates notch signaling pathway in angiogenesis induced by cerebral ischemia. Mol Cell Biochem 2012; 370: 45-51.
12. Li Q, He Q, Baral S, Mao L, Li Y, Jin H, et al. MicroRNA-493 regulates angiogenesis in a rat model of ischemic stroke by targeting MIF. FEBS J 2016; 283: 1720-1733.
13. Li Y, Mao L, Gao Y, Baral S, Zhou Y, Hu B. MicroRNA-107 contributes to post-stroke angiogenesis by targeting Dicer-1. Sci Rep 2015; 5: 13316.
14. Peltier HJ, Latham GJ. Normalization of microRNA expression levels in quantitative RT-PCR assays: identification of suitable reference RNA targets in normal and cancerous human solid tissues. RNA 2008;14: 844-852.
15. Wilfred BR, Wang WX, Nelson PT. Energizing miRNA research: a review of the role of miRNAs in lipid metabolism, with a prediction that miR-103/107 regulates human metabolic pathways. Mol Genet Metab 2007; 91:209-217.
16. Wilson R, Espinosa-Diez C, Kanner N, Chatterjee N, Ruhl R, Hipfinger C, et al. MicroRNA regulation of endothelial TREX1 reprograms the tumour microenvironment. Nat Commun 2016; 7: 13597.
17. Chen Z, Lai TC, Jan YH, Lin FM, Wang WC, Xiao H, et al. Hypoxia-responsive miRNAs target argonaute 1 to promote angiogenesis. J Clin Invest 2013; 123:1057-1067.
18. Greenberg DA, Jin K. Vascular endothelial growth factors (VEGFs) and stroke. Cell Mol Life Sci 2013; 70:1753-1761.
19. Bayne K. Revised Guide for the Care and Use of Laboratory Animals available. American Physiological Society. Physiologist 1996; 39:199, 208-11.
20. Yang M, Wang S, Hao F, Li Y, Tang H, Shi X. NMR analysis of the rat neurochemical changes induced by middle cerebral artery occlusion. Talanta 2012; 88: 136-144.
21. Shi H, Sun BL, Zhang J, Lu S, Zhang P, Wang H, et al. miR-15b suppression of Bcl-2 contributes to cerebral ischemic injury and is reversed by sevoflurane preconditioning. CNS Neurol Disord Drug Targets 2013; 12: 381-391.
22. Wang H, Zhang K, Zhao L, Tang J, Gao L, Wei Z. Anti-inflammatory effects of vinpocetine on the functional expression of nuclear factor-kappa B and tumor necrosis factor-alpha in a rat model of cerebral ischemia-reperfusion injury. Neurosci Lett 2014; 566: 247-251.
23. Wang R, Tian S, Wang HB, Chu DP, Cao JL, Xia HF, et al. MiR-185 is involved in human breast carcinogenesis by targeting Vegfa. FEBS Lett 2014; 588: 4438-4447.
24. Pedrioli DM, Karpanen T, Dabouras V, Jurisic G, van de Hoek G, Shin JW, et al. miR-31 functions as a negative regulator of lymphatic vascular lineage-specific differentiation in vitro and vascular development in vivo. Mol Cell Biol 2010; 30: 3620-3634.
25. Jiang G, Zhou R, He X, Shi Z, Huang M, Yu J, et al. Expression levels of microRNA-199 and hypoxia-inducible factor-1 alpha in brain tissue of patients with intractable epilepsy. Int J Neurosci 2016; 126:326-334.
26. Wang C, Ji B, Cheng B, Chen J, Bai B. Neuroprotection of microRNA in neurological disorders (Review). Biomed Rep 2014; 2: 611-619.
27. Jeyaseelan K, Lim KY, Armugam A. MicroRNA expression in the blood and brain of rats subjected to transient focal ischemia by middle cerebral artery occlusion. Stroke 2008; 39: 959-966.
28. Deng B, Du J, Hu R, Wang AP, Wu WH, Hu CP, et al. MicroRNA-103/107 is involved in hypoxia-induced proliferation of pulmonary arterial smooth muscle cells by targeting HIF-1beta. Life Sci 2016; 147: 117-124.
29. Kazemi-Lomedasht F, Behdani M, Pooshang Bagheri K, Habibi Anbouhi M, Abolhassani M, Khanahmad H, et al. Expression and purification of functional human vascular endothelial growth factor-a121; the most important angiogenesis factor. Adv Pharm Bull 2014; 4: 323-328.
30. Sun Y, Jin K, Xie L, Childs J, Mao XO, Logvinova A, et al. VEGF-induced neuroprotection, neurogenesis, and angiogenesis after focal cerebral ischemia. J Clin Invest 2003; 111: 1843-1851.
31. Margaritescu O, Pirici D, Margaritescu C. VEGF expression in human brain tissue after acute ischemic stroke. Rom J Morphol Embryol 2011; 52: 1283-1292.
32. Wang JX, Zhang XJ, Li Q, Wang K, Wang Y, Jiao JQ, et al. MicroRNA-103/107 regulate programmed necrosis and myocardial ischemia/reperfusion injury through targeting FADD. Circ Res 2015; 117: 352-363.
33. Vinciguerra A, Formisano L, Cerullo P, Guida N, Cuomo O, Esposito A, et al. MicroRNA-103-1 selectively downregulates brain NCX1 and its inhibition by anti-miRNA ameliorates stroke damage and neurological deficits. Mol Ther 2014; 22: 1829-1838.
34. Yang JP, Liu HJ, Liu XF. VEGF promotes angiogenesis and functional recovery in stroke rats. J Invest Surg 2010; 23: 149-155.
35. Sun J, Tao S, Liu L, Guo D, Xia Z, Huang M. miR1405p regulates angiogenesis following ischemic stroke by targeting VEGFA. Mol Med Rep 2016; 13: 4499-4505.
36. He QW, Li Q, Jin HJ, Zhi F, Suraj B, Zhu YY, et al. MiR-150 regulates poststroke cerebral angiogenesis via Vascular Endothelial Growth Factor in Rats. CNS Neurosci Ther 2016; 22: 507-517.