Objective: The mechanism of cerebral ischemia-reperfusion injury is closely related to inflammation, and miRNA-135b-5p may play a key role in the inflammation of cerebral ischemia injury. In this study we investigate the effect of miRNA-135b-5p through GSK3 β-TLR3 signal axis on inflammatory response in mice with cerebral ischemia-reperfusion injury. Methods: 48 mice were randomly divided into sham operation group (n = 12), model group (n = 12), negative control group (n = 12) and overexpression group (n = 12). After neurobehavioral score, the left brain of mice was killed and the water content of brain tissue was measured, the expression levels of TNF- α and IL-6 in brain tissue and serum were detected by ELISA method, the expression levels of TLR3, GSK3β and ERK protein in brain tissue were detected by Western Blot, and the mRNA expression levels of TLR3, GSK3 β and ERK in each group were detected by RT-PCR. Results: The neurobehavioral scores in the model group, negative control group and overexpression group were significantly higher than those in the sham operation group (P < 0.05), and the neurobehavioral scores in the overexpression group were significantly lower than those in the negative control group and model group (P < 0.05). The expression level of miRNA-135b-5p in brain tissue of mice in model group and negative control group was significantly lower than that in sham operation group, while the expression level of miRNA-135b-5p in overexpression group was significantly higher than that in other three groups. The expression levels of TNF- α and IL-6 in brain tissue and serum of mice in model group and negative control group were significantly higher than those in sham operation group, while the expression levels of TNF- α and IL-6 in overexpression group were significantly lower than those in model group and negative control group. The mRNA expression levels of TLR3, GSK3 β and ERK in the model group and negative control group were significantly higher than those in the sham operation group, while the mRNA expression levels of TLR3, GSK3 β and ERK in the overexpression group were significantly lower than those in the model group and negative control group. Conclusion: Overexpression of miRNA-135b-5p could inhibit the expression of GSK3β-TLR3 signal axis related genes and reduce the inflammatory response during cerebral ischemia-reperfusion injury, which may be a new strategy for the treatment of cerebral ischemia-reperfusion injury.

Qiang Duan, Silan Fan, Qian Wang, Chenhao Qi, Chao Li, Jie Huang, Chunxia Wei, Bo Wang, Xiaoqun Huang. miRNA-135b-5p Regulates GSK3β-TLR3 Signal Pathway to Inhibit Inflammatory Response with Cerebral Ischemia-Reperfusion Injury in Mice. International Journal of Frontiers in Medicine (2022), Vol. 4, Issue 10: 43-49. https://doi.org/10.25236/IJFM.2022.041007.

[1] Wu, Simiao et al. Stroke in China: advances and challenges in epidemiology, prevention, and management. The Lancet Neurology. 2019, Volume 18, Issue 4, 394 – 405.

[2] Wechsler LR. Intravenous thrombolytic therapy for acute ischemic stroke. N Engl J Med 2011; 364: 2138-46.

[3] Sims NR, Muyderman H. Mitochondria, oxidative metabolism and cell death in stroke. Biochim Biophys Acta 2010; 1802: 80-91.

[4] Wang P, Liang X, Lu Y, et al. MicroRNA-93 Downregulation Ameliorates Cerebral Ischemic Injury Through the Nrf2/HO-1 Defense Pathway. Neurochemical Research, 2016, 41(10): 1-9.

[5] Ping L, Fan J, Gao Y, et al. miR-135b-5p inhibits LPS-induced TNFα production via silencing AMPK phosphatase Ppm1e. Oncotarget, 2016, 7(47): 77978-77986.

[6] Qiang D, Wei S, Hua Y, et al. MicroRNA-135b-5p prevents oxygen-glucose deprivation and reoxygenation-induced neuronal injury through regulation of the GSK-3β/Nrf2/ARE signaling pathway. Archives of Medical Science Ams, 2018, 14(4): 735-744.

[7] Wang H, Brown J, Martin M. Glycogen synthase kinase 3: a point of convergence for the host inflammatory response. Cytokine, 2011, 53(2): 130-140.

[8] Beurel E, Michalek S M, Jope R S. Innate and adaptive immune responses regulated by glycogen synthase kinase-3 (GSK3). Trends in Immunology, 2010, 31(1): 24-31.

[9] Ko R, Park J H, Ha H, et al. Glycogen synthase kinase 3β ubiquitination by TRAF6 regulates TLR3-mediated pro-inflammatory cytokine production. Nature Communications, 2015, 6(1).

[10] Wang X, An F, Wang S et al. Orientin attenuates cerebral ischemia /reperfusion injury in rat model through the AQP-4 and TLR4 /NF-κB /TNF-α signaling pathway [J]. J Stroke Cerebrovasc Dis,2017,26( 10) : 2199-2214.

[11] Tu XK, Yang WZ, Chen JP et al. Curcumin inhibits TLR2 /4- NF-κB signaling pathway and attenuates brain damage in permanent focal cerebral ischemia in rats [J]. Inflammation,2014,37 (5): 1544-1551.

[12] Xiao G, Lyu M, Wang Y. Ginkgo Flavonol Glycosides or Ginkgolides Tend to Differentially Protect Myocardial or Cerebral Ischemia-Reperfusion Injury via Regu-lation of TWEAK-Fn14 Signaling in Heart and Brain [J]. Front Pharmacol,2019,10: 735-750.

[13] Yang Z, Weian C, Susu H et al. Protective effects of mangiferin on cerebral ischemia-reperfusion injury and its mechanisms [J]. Eur J Pharmacol,2015,771: 145-151.

[14] Xiaoyu Hu, Paul K. Paik, Janice Chen, et al. IFN-γ Suppresses IL-10 Production and Synergizes with TLR2 by Regulating GSK3 and CREB/AP-1 Proteins [J]. Immunity, 2006, 24(5):563.

[15] Beurel E, Michalek S M, Jope R S. Innate and adaptive immune responses regulated by glycogen synthase kinase-3 (GSK3) [J]. Trends in Immunology, 2010, 31(1):24.

[16] Ko R, Lee S Y. Glycogen synthase kinase 3β in Toll-like receptor signaling [J]. Bmb Reports, 2016, 49(6):305-310.

[17] Paul J, Naskar K, Chowdhury S, et al. TLR mediated GSK3 activation suppresses CREB mediated IL-10 production to induce a protective immune response against murine visceral leishmaniasis [J]. Biochimie.2014:107, 235-246.

[18] Li Y, Zhu J, Liu Y, et al. Glycogen Synthase Kinase 3beta Influences Injury Following Cerebral Ischemia/Reperfusion in Rats [J]. Int J Biol Sci 2016; 12: 518-31.

[19] He Y, Wang J, Wang J, et al. MicroRNA-135b regulates apoptosis and chemoresistance in colorectal cancer by targeting large tumor suppressor kinase 2. Am J Cancer Res, 2015, 5(4): 1382-1395.

[20] Hu K, Xie YY, Zhang C, et al. MicroRNA expression profile of the hippocampus in a rat model of temporal lobe epilepsy and miR-34a-targeted neuroprotection against hippocampal neurone cell apoptosis post-status epilepticus. BMC Neurosci 2012; 13: 115.

[21] Zhang Y, Xing H, Guo S, et al. MicroRNA-135b has a neuroprotective role via targeting of beta-site APP-cleaving enzyme 1. Exp Ther Med 2016; 12: 809-14.