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The Study of Therapeutic Effect and Mechanism of Mycophenolate Mofetil on Experimental Autoimmune Encephalomyelitis Rat

Author: TanXiaoZuo
Tutor: WangShunHe
School: Chongqing Medical University
Course: Pathology and Pathophysiology
Keywords: Experimental autoimmune encephalomyelitis Mycophenolate mofetil Multiple sclerosis Lymphocyte proliferation Adhesion molecule Matrix metalloproteinase
CLC: R744.3
Type: Master's thesis
Year: 2008
Downloads: 46
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Purpose of this research is the application mycophenolate mofetil (mycophenolate mofetil, MMF) animal model of multiple sclerosis (multiple sclerosis, MS) - experimental autoimmune encephalomyelitis (experimental autoimmune encephalomyelitis, EAE) in rats treated spleen lymphocyte proliferation and the central nervous system through the observation of clinical manifestations of experimental animals, and the central nervous system (central nervous system, CNS) pathological changes detected intercellular adhesion molecule -1 (intercellular adhesion molecule-1, ICAM-1) , the expression of vascular cell adhesion molecule -1 (vascular cell adhesion molecule-1, VCAM-1) and matrix metalloproteinase-9 (matrix metalloproteinase-9, MMP-9), mycophenolate mofetil treatment of EAE its mechanism of action, provided the experimental basis for the clinical treatment of multiple sclerosis. [Methods] (1) guinea pig spinal cord homogenate and complete Freund's adjuvant (complete freund's adjuvant, CFA) made of mixed emulsion as antigen supplemented intradermal injection of pertussis vaccine, Copy Wistar rats with acute EAE. (2) the experimental animals were randomly divided into three groups, normal control group, the EAE model group and the MMF treatment group, n = 12. 10 days after immunization, MMF treatment group mycophenolate mofetil 30mg · kg -1 · d -1 dose intragastrically 1 week of treatment, EAE model The group with normal saline instead. (3) immune laboratory animals after daily observation of changes in body weight and clinical symptom score; 16 days after immunization, 3 groups from each part of the rats were killed. HE staining, Luxol Fast Blue-HE myelin staining, by Bielschowsky the axon silver staining and electron microscopy observation of the central nervous system pathological changes; lymphocyte proliferation ability of CCK-8 assay; immunohistochemical method for the detection of the central nervous system of ICAM- 1, VCAM-1 and MMP-9 protein expression. Observed groups remaining rats were sacrificed on day 30, to the immune daily still the weight weighing and clinical symptom scores, in order to understand the whole course of the disease changes. [Results] (1) modeling rats clinical manifestations, pathological changes are in line with the characteristics of acute EAE lesions. Peak ratings of clinical symptoms (2) EAE model rats (2.83 ± 1.27), weight loss maximum value (34.58 ± 13.47) g, the duration of (16.33 ± 1.15) d. MMF-treated rats reduce clinical symptoms, symptoms peak score (1.75 ± 1.12), the highest weight loss (24.67 ± 8.42) g, and the duration of (11.67 ± 1.53) d, compared with the EAE group differences were statistically significant (P lt; 0.05). HE staining showed that EAE group rat brain, the cerebellum, spinal cord inflammatory cell infiltration score were (2.50 ± 0.55), (3.50 ± 0.84), (3.83 ± 0.41), the corresponding parts in the MMF treatment scores were (1.5 ± 0.55), (2.00 ± 0.63), (2.17 ± 0.98), the difference was statistically significant (P lt; 0.05). Luxol Fast Blue-HE myelin staining the EAE group rat brain, the cerebellum, and spinal cord demyelination score were (1.33 ± 0.52), (2.00 ± 0.63), (2.17 ± 0.75), the corresponding parts of MMF treatment group scores were (0.83 ± 0.41), (1.00 ± 0.63), (1.17 ± 0.41), the difference was statistically significant (P lt; 0.05). The Bielschowsky the axon silver staining Display MMF spinal cord axonal injury score (1.00 ± 0.63), compared with the EAE group (1.83 ± 0.75), the difference was statistically significant (P lt; 0.05). Electron microscopy results showed ultrastructural changes of the myelin and axons of MMF treatment group compared with EAE model group was significantly reduced. (3) EAE model in spleen lymphocyte stimulation index (2.48 ± 0.29), the stimulation index of MMF treatment group reduced to (1.68 ± 0.24), the difference was statistically significant (p lt; 0.05). The the EAE model group in the rat central nervous system of ICAM-1, VCAM-1 positive number of blood vessels and expression of MMP-9-positive cell rate, respectively (43.61 ± 5.05), (41.28 ± 4.33), (46.93 ± 6.50)%, MMF group indicators expression (10.89 ± 1.76), (11.39 ± 2.08), (20.58 ± 3.34)%, between the two groups of indicators compare the differences were statistically significant (p lt; 0.05). [Conclusion] (1) copy the acute EAE model in this study is successful, stable and reliable. (2) mycophenolate mofetil the acute EAE has obvious therapeutic effect, can effectively improve the clinical manifestations of EAE, reduce inflammatory cell infiltration of the central nervous system, reduce the damage of the myelin sheath of demyelination and axon. (3) The mechanism of action of mycophenolate mofetil treatment of EAE is multifaceted, not only with the suppression of the immune response may also play its therapeutic effect by reducing EAE central nervous system, ICAM-1, VCAM-1 and MMP-9 expression .

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