| Objective and significance:Experimental autoimmune encephalomyelitis (EAE) was an active T cell mediated organ specific autoimmune disease. Clinical characteristics were inflammatory cells infiltrating around blood vessel and central nerve demyelination which had similar teatures with multiple sclerosis (MS) in clinical symptom, biochemical indicator and pathology. So EAE was attributed as favorable animal models for studying MS. Nowadays, many scholars made EAE model by using susceptive animals to interpret pathogenesis on MS and EAE, offer experimental evidence on detecting pathogenetic condition, prevent relapse, select advisable plan and bolt out new methods and new medicine. Therefore, making stable and dependable EAE model will be the basis of developing a series of EAE researches. Some researchers had discovered that mesenchymal stem cells which from fat were infused into animal models on the prophase of EAE, which could ease inflammation of spinal cord and change demyelination and detect the these adult stem cells (ASCs) in lymphatic organ and central nervous system. But there weren't related to exact mechanism of ASCs. By the deep-going researches on immunological characteristics of MSCs, we believe that MSCs will be got general applications in autoimmune disease (AID). This study was aimed at observing the effects of the process of murine mesenchymal stem cell C3H10 transplanting into EAE mice, and then to give the basis of experiments for researching the therapy of autoimmune disease such as MS.Methods:In the first part of the study, twenty-four female C57BL/6J mice were randomly divided into three groups: non-intervene group, C3H10 transplantation group and normal group. The emulsionized antigen adjuvant was obtained by mixation of myelin oligodendrocyte glycoprotein (MOG35-55) and complete Freund's adjuvant (CFA) and then immunized the mice of the experimental group. Bordetella pertussis toxin (PT) was injected by tail vein on day 0 and after 24 fours while control group with PBS treatment. On the eighth day after immunized injecting the C3H10 cells which were stained by CFSE through caudal vein, the clinical manifestation of three groups were rated and analyzed. The spinal tissue of experiment group with apparent clinical manifestation and control group were sliced and HE, Luxol Fast-Blue stained in order to identify that the animal model was successful or not. And then sign and pathologic changes had happened to apresent apparent therapeutic actions or not after transplanting C3H10. In the second part of the study, we made the cryosections with fresh spinal cord to handle the mice in transplantation of C3H10 group, and searched the C3H10 which were got into spinal cord or not trough fluorescence microscope.Results:(1) The changes of animals'action: From the tenth day after immuning, non-intervention group emerges the clinical symptom, peaking in the 14-18day, average neurological score is about 2±0.53(p<0.05); In transplantation of C3H10 group, there are two mice's neurological score is about 1 in the 14th day and the 18th day respectively; Normal group didn't emerge any clinical symptom (0 score), keeping 35 days continuous observation. (2) The changes of animals'weight: The weights of non-intervention group keep decreasing by degrees, the weights is about 18.93±2.12g in the 14th day, 18.65±2.04g in the 22th day; The weights of transplantation of C3H10 group had the trend of ascent, up to 18.7±0.93g and keep the weight on 19.26±0.58g after making model. It indicated that the two groups existed statistical difference (P<0.05). (3) Pathological changes: Inflammatory cells infiltrated around brain and spinal cord in non-intervention group, with exhibited cuffing or demyelinating changes. While transplantation of C3H10 group and normal group didn't emerge these clinical changes. (4) There had fluorescence which liked the appearance of C3H10 from the mice's spinal cord in transplantation of C3H10 group.Conclusions:(1) The animal model of EAE was successfully constructed whose clinical symptoms and pathological changes were consistent with recrudescent and non-relieved EAE and the model was stable and reliable.(2) Transplantation of murine Mesenchyma1 Stem Cells C3H10 into EAE mice could decrease and delay the development of disease, and then present apparent therapeutic action. |
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