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Study on the Treatment of Hydroxyfasudil and Its Influence on Expression of ROCK-Ⅱ, INF-γ, IL-10 and IL-17 in Experimental Autoimmune Encephalomyelitis Mice

Author: DuanHuiJie
Tutor: MaCunGen
School: Shanxi Medical
Course: Neurology
Keywords: Experimental autoimmune encephalomyelitis Fasudil hydrochloride ROCK-II Lymphocyte proliferation IFN-γ IL-17 IL-10
CLC: R744.51
Type: Master's thesis
Year: 2010
Downloads: 112
Quote: 0
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Abstract


Objective: the experimental application of hydrochloric acid fasudil (Fasudil hydrochloride) intervention in the treatment of myelin oligodendrocytes glial glycoprotein 35-55 peptides (myelin oligodendrocyte glycoprotein peptides. 35-55 MOG35-55) induced C57BL / 6 mice experiments autoimmune encephalomyelitis (experimental autoimmune encephalomyelitis, EAE) model, observed a group of clinical symptoms, pathological changes detected expression of ROCK-Ⅱ in the brains of mice in each group, and the spleen lymphocyte proliferation and cell supernatants of IFN -γ, IL-10, IL-17 cytokine secretion changes, to study the role of Rho / Rock signaling pathway in the pathogenesis of EAE observed treatment effect of Rho kinase inhibitor hydrochloric fasudil EAE its on ROCK- II splenic lymphocyte proliferative activity and cytokine secretion, and to explore the occurrence of EAE development mechanism and application of hydrochloric acid fasudil treatment of EAE mechanism of action, to provide experimental evidence for the treatment of multiple sclerosis. Methods: 20 female C57BL / 6 mice (18-20g) 60 Zhi divided into three groups, according to the average weight EAE group, the treatment group and the adjuvant group MOG35-55 induced EAE model, the adjuvant group was replaced by normal saline. Treatment group in the first 6 days after immunization by intraperitoneal injection of 50mg/kg · d hydrochloric fasudil liquid, for 14 consecutive days, EAE group and the adjuvant group with normal saline as a control. General observation and comparison of mice in each group, body weight changes, clinical manifestations, and symptom scores. After vaccination 18-20 days each group removed eight mice, aseptic isolation of the spleen lymphocytes, the cell suspension was prepared. Groups spleen lymphocytes proliferation was detected by MTT detected by ELISA and compared among the groups lymphocyte culture supernatant cytokines TNF-γ, IL-17, IL-10 levels. Anesthesia 18-20 days after immunization, the animals were sacrificed, dissecting the brain and spinal cord, HE staining, the myelin staining observed pathological changes, the Western blot method to detect the expression of ROCK-Ⅱ. Using SPSS13.0 software analysis and processing of the data obtained. The results: 1. Use of MOG35-55 peptides induced C57BL / 6 mouse EAE model, HE staining showed significant inflammatory infiltration in EAE mice spinal cord, myelin staining apparent demyelination. Compared with EAE symptoms treatment group late light performance, morbidity, weight change, mean maximum symptom score differences were statistically significant (P lt; 0.05), inflammatory cell infiltration and demyelination change significantly reduced. The adjuvant group no disease, weight increasing trend. 2.EAE group spleen lymphocyte proliferation significantly (P lt; 0.01); and lymphocyte supernatant of INF-γ, IL-17 secretion increased (P lt; 0.05). Fasudil hydrochloride spleen lymphocyte proliferative response was significantly inhibited (P lt; 0.01), and significant inhibition of IFN-γ, IL-17 secretion (P lt; 0.05), IL-10 secretion obvious role (P lt; 0.05). 3.EAE group, the treatment group and adjuvant mice brain were ROCK-II expression. EAE group ROCK-Ⅱ expression is higher as compared to the adjuvant group (P lt; 0.05), compared to the treatment group and the EAE group was significantly lower (P lt; 0.05). Conclusion: fasudil hydrochloride can effectively treat mice of EAE, reduce the incidence, remission of clinical symptoms, relieve inflammatory infiltration and demyelination. Fasudil hydrochloride can inhibit T cell proliferation and INF-γ secretion of pro-inflammatory cytokines, promote anti-inflammatory cytokines such as IL-10 secretion fasudil effectively suppress mice EAE possible mechanism. 3 inhibit activation of ROCK-Ⅱ is another possible mechanism of the effective suppression of EAE in mice.

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