Toll‑Like receptor 4 promotes the phosphorylation of CRMP2 via the activation of Rho‑kinase in MCAO rats

Li,Xue‑Bo; Ding,Ming‑Xia; Ding,Chun‑Li; Li,Liang‑Liang; Feng,Jinzhou; Yu,Xiao‑Jun

doi:10.3892/mmr.2018.8968

Toll‑Like receptor 4 promotes the phosphorylation of CRMP2 via the activation of Rho‑kinase in MCAO rats

Authors:
- Xue‑Bo Li
- Ming‑Xia Ding
- Chun‑Li Ding
- Liang‑Liang Li
- Jinzhou Feng
- Xiao‑Jun Yu
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Affiliations: Department of Forensic Medicine, Shantou University Medical College, Shantou, Guangdong 515041, P.R. China, Department of Obstetrics and Gynecology, The Second Hospital of Shandong University, Jinan, Shandong 250014, P.R. China, Key Laboratory of Evidence Identification in Universities of Shandong, Shandong University of Political Science and Law, Chongqing 400016, P.R. China, Department of Neurology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, P.R. China
Published online on: May 3, 2018 https://doi.org/10.3892/mmr.2018.8968
Pages: 342-348
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The mechanism associated with Toll‑like receptor 4 (TLR4) in neurological injury remains unclear. The aim of the present study was to investigate the pathology of TLR4 in middle cerebral artery occlusion (MCAO)/reperfusion rat models via the regulation of collapsin response mediator protein 2 (CRMP2) phosphorylation. The modified neurological severity score (mNSS) was applied to assess neurological recovery. Immunofluorescence and western blotting were used to detect the protein expressions of TLR4, Rho‑associated protein kinase 2 (ROCK‑II) and CRMP2 following the intracerebroventricular administration of TLR4‑specific agonist, lipopolysaccharide (LPS) and TLR4‑neutralizing antibody, the ROCK‑II specific inhibitor Y‑27632 or LPS+Y‑27632 30 min prior to MCAO. The expression levels of TLR4 and the phosphorylation of CRMP2 significantly increased in response to LPS‑mediated induction and/or MCAO; however, they were reversed by treatment with LPS+TLR4‑neutralizing antibody. Y‑27632 decreased the expression of ROCK‑II and phosphorylated (p)‑CRMP2, and suppressed the increased ROCK‑II and p‑CRMP2 induced by LPS; however, no effect on the levels of TLR4 expression was observed. The neurological function as measured by mNSS score was reduced in the LPS group when compared with the MCAO group, whereas the LPS+Y‑27632 group reversed the reduced neurological function at 7 and 14 days post‑MCAO. The results of the present study suggested that TLR4 may promote the phosphorylation of CRMP2 via the activation of ROCK‑II in MCAO rats, which further characterizes the pathological mechanism of TLR4 in stroke, and that modulation of TLR4 could be a potential target to limit secondary post‑stroke brain damage.

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1	Mensah GA, Norrving B and Feigin VL: The global burden of stroke. Neuroepidemiology. 45:143–145. 2015. View Article : Google Scholar : PubMed/NCBI
2	Herson PS and Traystman RJ: Animal models of stroke: Translational potential at present and in 2050. Future Neurol. 9:541–551. 2014. View Article : Google Scholar : PubMed/NCBI
3	Sierra C, Coca A and Schiffrin EL: Vascular mechanisms in the pathogenesis of stroke. Curr Hypertens Rep. 13:200–207. 2011. View Article : Google Scholar : PubMed/NCBI
4	Xu X, Wen Z, Zhao N, Xu X, Wang F, Gao J, Jiang Y and Liu X: MicroRNA-1906, a Novel regulator of toll-like receptor 4, ameliorates ischemic injury after experimental stroke in mice. J Neurosci. 37:10498–10515. 2017. View Article : Google Scholar : PubMed/NCBI
5	Tang SC, Arumugam TV, Xu X, Cheng A, Mughal MR, Jo DG, Lathia JD, Siler DA, Chigurupati S, Ouyang X, et al: Pivotal role for neuronal Toll-like receptors in ischemic brain injury and functional deficits. Proc Natl Acad Sci USA. 104:13798–13803. 2007. View Article : Google Scholar : PubMed/NCBI
6	Tang SC, Yeh SJ, Li YI, Wang YC, Baik SH, Santro T, Widiapradja A, Manzanero S, Sobey CG, Jo DG, et al: Evidence for a detrimental role of TLR8 in ischemic stroke. Exp Neurol. 250:341–347. 2013. View Article : Google Scholar : PubMed/NCBI
7	Olson JK and Miller SD: Microglia initiate central nervous system innate and adaptive immune responses through multiple TLRs. J Immunol. 173:3916–3924. 2004. View Article : Google Scholar : PubMed/NCBI
8	Lehnardt S, Lehmann S, Kaul D, Tschimmel K, Hoffmann O, Cho S, Krueger C, Nitsch R, Meisel A and Weber JR: Toll-like receptor 2 mediates CNS injury in focal cerebral ischemia. J Neuroimmunol. 190:28–33. 2007. View Article : Google Scholar : PubMed/NCBI
9	Fukata Y, Itoh TJ, Kimura T, Ménager C, Nishimura T, Shiromizu T, Watanabe H, Inagaki N, Iwamatsu A, Hotani H and Kaibuchi K: CRMP-2 binds to tubulin heterodimers to promote microtubule assembly. Nat Cell Biol. 4:583–591. 2002. View Article : Google Scholar : PubMed/NCBI
10	Yoshimura T, Kawano Y, Arimura N, Kawabata S, Kikuchi A and Kaibuchi K: GSK-3beta regulates phosphorylation of CRMP-2 and neuronal polarity. Cell. 120:137–149. 2005. View Article : Google Scholar : PubMed/NCBI
11	Gim SA, Sung JH, Shah FA, Kim MO and Koh PO: Ferulic acid regulates the AKT/GSK-3β/CRMP-2 signaling pathway in a middle cerebral artery occlusion animal model. Lab Anim Res. 29:63–69. 2013. View Article : Google Scholar : PubMed/NCBI
12	Ye Y, Xu H, Zhang X, Li Z, Jia Y, He X and Huang JH: Association between toll-like receptor 4 expression and neural stem cell proliferation in the hippocampus following traumatic brain injury in mice. Int J Mol Sci. 15:12651–12664. 2014. View Article : Google Scholar : PubMed/NCBI
13	Julian L and Olson MF: Rho-associated coiled-coil containing kinases (ROCK): Structure, regulation, and functions. Small GTPases. 5:e298462014. View Article : Google Scholar : PubMed/NCBI
14	Yoneda A, Multhaupt HA and Couchman JR: The rho kinases I and II regulate different aspects of myosin II activity. J Cell Biol. 170:443–453. 2005. View Article : Google Scholar : PubMed/NCBI
15	National Research Council, . Guide for the care and use of laboratory animals. 8th edition. National Academy Press; Washington, DC: 2011, https://grants.nih.gov/grants/olaw/guide-for-the-care-and-use-of-laboratory-animals.pdf PubMed/NCBI
16	Arumugam TV, Salter JW, Chidlow JH, Ballantyne CM, Kevil CG and Granger DN: Contributions of LFA-1 and Mac-1 to brain injury and microvascular dysfunction induced by transient middle cerebral artery occlusion. Am J Physiol Heart Circ Physiol. 287:H2555–H2560. 2004. View Article : Google Scholar : PubMed/NCBI
17	Longa EZ, Weinstein PR, Carlson S and Cummins R: Reversible middle cerebral artery occlusion without craniectomy in rats. Stroke. 20:84–91. 1989. View Article : Google Scholar : PubMed/NCBI
18	Xiong X, Barreto GE, Xu L, Ouyang YB, Xie X and Giffard RG: Increased brain injury and worsened neurological outcome in interleukin-4 knockout mice after transient focal cerebral ischemia. Stroke. 42:2026–2032. 2011. View Article : Google Scholar : PubMed/NCBI
19	Iwasa T, Matsuzaki T, Tungalagsuvd A, Munkhzaya M, Kawami T, Kato T, Kuwahara A, Yasui T and Irahara M: Effects of ovariectomy on the inflammatory responses of female rats to the central injection of lipopolysaccharide. J Neuroimmunol. 277:50–56. 2014. View Article : Google Scholar : PubMed/NCBI
20	Ma Y, Zhang X, Bao H, Mi S, Cai W, Yan H, Wang Q, Wang Z, Yan J, Fan GC, et al: Toll-like receptor (TLR) 2 and TLR4 differentially regulate doxorubicin induced cardiomyopathy in mice. PLoS One. 7:e407632012. View Article : Google Scholar : PubMed/NCBI
21	Gong G, Bai S, Wu W, Hu L, Liu Y, Niu J, Dai X, Yin L and Wang X: Lrg participates in lipopolysaccharide preconditioning-induced brain ischemia injury via TLR4 signaling pathway. J Mol Neurosci. 54:20–26. 2014. View Article : Google Scholar : PubMed/NCBI
22	Rikitake Y, Kim HH, Huang Z, Seto M, Yano K, Asano T, Moskowitz MA and Liao JK: Inhibition of rho kinase (ROCK) leads to increased cerebral blood flow and stroke protection. Stroke. 36:2251–2257. 2005. View Article : Google Scholar : PubMed/NCBI
23	Sugawara T, Ayer R, Jadhav V and Zhang JH: A new grading system evaluating bleeding scale in filament perforation subarachnoid hemorrhage rat model. J Neurosci Methods. 167:327–334. 2008. View Article : Google Scholar : PubMed/NCBI
24	Kawai T and Akira S: The role of pattern-recognition receptors in innate immunity: Update on toll-like receptors. Nat Immunol. 11:373–384. 2010. View Article : Google Scholar : PubMed/NCBI
25	Medzhitov R and Janeway CA Jr: Innate immunity: The virtues of a nonclonal system of recognition. Cell. 91:295–298. 1997. View Article : Google Scholar : PubMed/NCBI
26	Doi Y, Mizuno T, Maki Y, Jin S, Mizoguchi H, Ikeyama M, Doi M, Michikawa M, Takeuchi H and Suzumura A: Microglia activated with the toll-like receptor 9 ligand CpG attenuate oligomeric amyloid {beta} neurotoxicity in in vitro and in vivo models of Alzheimer's disease. Am J Pathol. 175:2121–2132. 2009. View Article : Google Scholar : PubMed/NCBI
27	Hensley K, Venkova K, Christov A, Gunning W and Park J: Collapsin response mediator protein-2: An emerging pathologic feature and therapeutic target for neurodisease indications. Mol Neurobiol. 43:180–191. 2011. View Article : Google Scholar : PubMed/NCBI
28	Brown M, Jacobs T, Eickholt B, Ferrari G, Teo M, Monfries C, Qi RZ, Leung T, Lim L and Hall C: Alpha2-chimaerin, cyclin-dependent Kinase 5/p35, and its target collapsin response mediator protein-2 are essential components in semaphorin 3A-induced growth-cone collapse. J Neurosci. 24:8994–9004. 2004. View Article : Google Scholar : PubMed/NCBI
29	Arimura N, Ménager C, Kawano Y, Yoshimura T, Kawabata S, Hattori A, Fukata Y, Amano M, Goshima Y, Inagaki M, et al: Phosphorylation by rho kinase regulates CRMP-2 activity in growth cones. Mol Cell Biol. 25:9973–9984. 2005. View Article : Google Scholar : PubMed/NCBI
30	Wang T, Wu X, Yin C, Klebe D, Zhang JH and Qin X: CRMP-2 is involved in axon growth inhibition induced by RGMa in vitro and in vivo. Mol Neurobiol. 47:903–913. 2013. View Article : Google Scholar : PubMed/NCBI