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Research the Effects of Tacromulis on Rats with Experimental Autoimmune Encephalomyelitis

Author: ShiHaiBo
Tutor: WangShunHe
School: Chongqing Medical University
Course: Pathology and Pathophysiology
Keywords: Experimental autoimmune encephalomyelitis Tacrolimus Multiple sclerosis Growth associated protein Rats
CLC: R744.51
Type: Master's thesis
Year: 2009
Downloads: 59
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Abstract


[Objective] In this study, immune inhibitors tacrolimus (FK506, tacromulis) treatment of multiple sclerosis in animal models --- experimental allergic encephalomyelitis (Experimental Autoimmune Encephalomyelitis EAE) rats, by observing the experiment pathological changes of the animal clinical manifestations and the central nervous system (central nervous sysyem, CNS), peripheral blood T lymphocytes CD3, CD4, CD8, and NK cell ratio neuron growth cone of the major protein components phosphate growth-related protein-43 (Gap -43 Growth Associated Protein 43) investigate the expression the tacrolimus treatment effect and mechanism of EAE and prednisolone as compared with the traditional treatment methods, provided the experimental basis for the clinical treatment of multiple sclerosis. [Methods] (1) with guinea pig spinal cord homogenate and complete Freund's adjuvant (complete freund's adjuvant, CFA) made from a mixture of antigen intradermal injection of pertussis vaccine, copy the SD rat animal model of multiple sclerosis acute EAE. (2) the experimental animals were randomly divided into five groups, the normal control group (n = 5), the EAE model group (n = 10), FK506A group (n = 10), FK506B group (n = 10), prednisolone control group (n = 10). Prednisolone control group 10 days after immunization began treatment, FK506A group tacrolimus group 1mg/kg/d, FK506B tacrolimus 0.5mg/kg/d dose continuous intraperitoneal injection of 10 days, 10mg/kg/d dose intraperitoneal injection of 10 days, the saline instead the EAE group with the same amount. (3) changes in weight of the experimental animals were observed daily after immunization and clinical score; 20 days after immunization, mining rats tail vein EDTA anticoagulant, as measured by flow cytometry of T lymphocytes CD3, CD4, CD8 8, and NK cells ratio changes; 30 days after immunization, all rats of each group were sacrificed, HE, Luxol Fast Blue-HE myelin staining by Bielschowsky axons Silver staining observed pathological changes of the central nervous system; GAP-43 immunohistochemistry chemical staining Morphological evidence of axonal regeneration in the central nervous system. Results (1) copy the model of the rat clinical manifestations, pathological changes are in line with the characteristics of acute EAE lesions. Peak score of clinical symptoms (2) EAE control rats (3.02 ± 0.98), reduce the maximum weight (39.62 ± 13.64) g, onset time (10.21 ± 1.08) d after immunization, clinical course continued (16.52 ± 0.83 ) d. FK506A rats clinical symptoms peak ratings for (1 .. 21 ± 0.31), reduce the maximum weight (12.52 ± 5.26) g, onset time (14.61 ± 1.74) d after immunization, clinical course continued (10.34 ± 0.61) d. FK506B rats clinical symptoms peak score (1.52 ± 0.36), reduce the maximum weight (11.34 ± 5.19) g, immunity after the onset time (14.08 ± 1.39) d, the clinical course of the disease continued (10.91 ± 1.21) d. prednisolone clinical peak ratings of control rats (1.67 ± 0.22), body weight to reduce the maximum value (-26.12 ± 7.34), time of onset (10.41 ± 0.81) d after immunization, clinical course continued (18.77 ± 0.56) d. HE staining showed that EAE group rat brain, the cerebellum, spinal cord inflammatory cell infiltration score (2.61 ± 0.47), (3.17 ± 0.29), (3.86 ± 0.44), FK506A rat brain, the cerebellum, spinal cord inflammatory cells infiltration scores were (1.31 ± 0.54), (1.94 ± 0.27), (2.08 ± 0.23), FK506B rat brain, the cerebellum, spinal cord inflammatory cell infiltration score (1.27 ± 0.69), (1.87 ± 0.35), (2.12 ± 0.76), prednisolone control rats brain, the cerebellum, spinal cord inflammatory cell infiltration score were (1.12 ± 0.41), (2.31 ± 0.14), (2.29 ± 0.58). Luxol Fast Blue-HE myelin staining the EAE group rat brain, the cerebellum, and spinal cord demyelination score were (1.59 ± 0.83), (2.07 ± 0.74), (2.21 ± 0.51), FK506A group rat brain , cerebellum, spinal cord demyelination score were (0.86 ± 0.31), (1.29 ± 0.64), (1.44 ± 0.52), FK506B rat brain, the cerebellum, spinal cord demyelination score (0.87 ± 0.17), (1.34 ± 0.48), (1.51 ± 0.72), prednisolone control rats brain, the cerebellum, and spinal cord demyelination scores were (0.81 ± 0.33), (1.27 ± 0.58), (1.17 ± 0.28). The Bielschowsky the axon silver staining Display EAE spinal cord axonal injury score (1.91 ± 0.64), FK506A group showed spinal cord axonal injury score (1.17 ± 0.26), FK506B group showed spinal cord axonal injury score (1.31 ± 0.37), poured Nepal rated prednisolone spinal axon damage control group (1.06 ± 0.14). CD3 T lymphocyte ratio in peripheral blood of EAE group, FK506A group the, FK506B group, prednisolone control group (67.22 ± 1.21), (58.17 ± 0.96), (60.28 ± 2.13), (71.06 ± 3.83). Rat peripheral blood CD4 T lymphocyte ratio EAE group, FK506A group the, FK506B group, prednisolone control group (51.27 ± 2.13), (41.34 ± 1.41), (44.12 ± 2.69), (55.21 ± 4.78). EAE group, FK506A group the, FK506B group, prednisolone control rats peripheral blood CD8 T lymphocyte ratio (28.16 ± 1.53), respectively, (14.43 ± 0.74) (13.2 ± 1.17), (26.34 ± 1.72), the rat peripheral NK EAE group, FK506A group the, FK506B group, prednisolone control group cell ratio (8.52 ± 1.16), respectively (1.07 ± 0.41), (1.89 ± 0.21), (7.62 ± 0.49), EAE group CD4 / CD8 ratio (1.82 ± 0.26), FK506A CD4 / CD8 ratio (2.61 ± 0.21), FK506B CD4 / CD8 ratio (2.84 ± 0.51), prednisolone control group CD4 / CD8 ratio (2.09 ± 0.38). EAE group FK506A group the, FK506B group, prednisolone control group rat central nervous system, GAP-43 immunohistochemical staining positive cells were (195.96 ± 21.51), (454.52 ± 17.63), (397.19 ± 19.09 ), (211.61 ± 12.51). [Conclusion] (1) copy the acute EAE model in this study is successful, stable and reliable. (2) tacrolimus Secretary for the treatment of acute EAE, has effectively improved the the EAE clinical manifestations, alleviate the inflammation of the central nervous system damage, demyelination and axon damage. (3) after the withdrawal of the Secretary treatment group, no clinical symptoms recurred tacrolimus, tacrolimus has significantly delayed EAE clinical onset time and shorten the clinical course of time. (4) tacrolimus Division of the treatment of EAE rats may be related to the inhibition of T cell immunity, may also promote axonal regeneration of the central nervous system to exert its therapeutic role.

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