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The Effect of Fasudil on Expression of Rock Ⅱ in Spinal Cords in Experimental Autoimmune Encephalomyelitis Mice

Author: LiYanXiang
Tutor: MaCunGen
School: Shanxi Medical
Course: Neurology
Keywords: Experimental autoimmune encephalomyelitis Multiple sclerosis Rho kinase Rho / Rock signaling pathway Fasudil
CLC: R744.5
Type: Master's thesis
Year: 2011
Downloads: 22
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Abstract


Objective: Rho / Rho kinase (Rock) signaling pathway is a ubiquitous organism signal transduction pathways involved in regulation of cell proliferation, adhesion, migration, etc., are believed to control cell behavior upstream signaling molecules. Recent studies have found that many neuropathological Rock activation state, and the application of Rock inhibitors can improve the clinical symptoms and pathological conditions. The experimental application Rho kinase inhibitor Fasudil treatment C57BL / 6 mice EAE, clinical efficacy and potential immunological mechanisms, aimed at exploring the pathogenesis of EAE and application of Fasudil treatment of EAE mechanism for EAE and MS immunopathology mechanisms to provide new theoretical support, but also for the future clinical applications Fasudil adjuvant treatment of CNS autoimmune diseases MS provide experimental evidence. Methods: Female C57BL / 6 mice were randomly divided into EAE group 20, Fasudil intervention group 20, adjuvant group 20. EAE and Fasudil intervention group with myelin oligodendrocyte glycoprotein 35-55 (MOG35-55) were immunized with established EAE model, adjuvant group replaced by normal saline MOG35-55 EAE group as a control. Fasudil intervention group in the first seven days after immunization by intraperitoneal injection Fasudil (50mg/kg/d), continuous treatment for 14 days. EAE group and the adjuvant group given normal saline as a control. Immunized animals in each group observed the day after the body weight change and incidence, according to the international standard assessment of clinical symptoms. 4 weeks after immunization the animals were killed at the mouse spinal cord lumbar enlargement, HE staining, Luxol Fast Blue staining and indirect immunofluorescence staining to observe pathological changes of mice in each group and the Rock-Ⅱ expression. Experimental data were analyzed using SPSS13.0 software. Results: 1. Use MOG35-55 peptide successfully induced C57BL / 6 mice EAE model. Incidence of EAE group 100% (n = 20), mean onset time was 13.35 ± 1.31 days, the peak of the average symptom score 3.80 ± 0.78, the peak of the average weight 15.75 ± 0.81. In the first 12 days after the onset began after another obvious clinical symptoms, the incidence of varying severity, the first performance of eating less, less active, and the fur is not smooth, weight loss, weakness tail drooping, with exacerbations occur hindlimb paralysis on one side, and further development of bilateral hindlimb paralysis, can not be independent stand, accompanied by severe forelimb paralysis, the highest score of 4 points. Fasudil intervention group incidence of 35% (n = 20), compared with EAE group decreased; mean onset time of 15.55 ± 1.50 days, compared with EAE group was significantly delayed (P lt; 0.01); peak of the mean symptom score 2.24 ± 0.26, compared with EAE group decreased (P lt; 0.05); peak average weight of 19.56 ± 0.66, compared with EAE significantly different (P lt; 0.05). This 4 Comparison of clinical indicators, Fasudil intervention group and EAE group were statistically significant. Adjuvant group, no disease, body weight continued to increase. 2.HE staining showed that, EAE mice significant inflammatory cell infiltration, Fasudil intervention group and mild inflammatory infiltration adjuvant group, EAE group, Fasudil intervention group and the adjuvant group Okuda inflammatory cell infiltration score was significantly different. EAE group of inflammatory cell infiltration score 3.25 ± 0.56, Fasudil intervention group, 2.13 ± 0.44, adjuvant group 1.38 ± 0.78. Fasudil EAE group and intervention group, EAE group and the adjuvant group were statistically significant (P lt; 0.05). Luxol Fast Blue staining, EAE group demyelination than Fasudil intervention group and adjuvant were more obvious, and with the severity of the disease were positively correlated. 3. Rock-Ⅱ in EAE group, Fasudil spinal intervention group and adjuvant group were expressed, and the expression of the disease in mice and clinical severity score consistent with endothelial cells in EAE group clearly expressed. Observe the same field within the spinal cord Rock-Ⅱ positive cells, EAE mice was 31.78 ± 1.04, Fasudil intervention group 20.22 ± 1.56, adjuvant group was 18.50 ± 1.39, EAE group and Fasudil intervention group, EAE group and the adjuvant group phase There were significant differences (P lt; 0.01). The results show that Fasudil Rock-Ⅱ intervention group was significantly lower than EAE group. Conclusions: 1. Antigen emulsion with MOG35-55 immunized C57BL / 6 mice, can be successfully established EAE model. 2.Rho/Rock signaling pathway with EAE pathogenesis. 3.Rock inhibitor - Fasudil can reduce the incidence of EAE, alleviate EAE clinical symptoms, reduce the inflammatory cell infiltration and demyelination. 4. Rock inhibitors - Fasudil reduced EAE mouse spinal cord and vascular endothelial cells Rock-Ⅱ expression may be the mechanism of immune suppression of EAE.

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CLC: > Medicine, health > Neurology and psychiatry > Neurology > Spinal cord disease > Demyelinating disease
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