The Variation Of Regulatory CD4~+CD25~+Foxp3~+Tcell,PCNA Of Thymus In Different Courses Of EAE Group And The Effection Of Alpha Lipoic Acid

Objective:Multiple sclerosis(MS) is one kind of central nervous system demyelinating diseases, which is mediated by T cell mainly.Thymus is the important central immune organ, which is important to produce and progresse T cell. Experimental autoimmune encephalomyelitis is an classical animal model to research MS. The a-lipoic is one kind of strong antioxidant which can relieving the symptom of EAE according to the study before. In our experiments, EAE was induced by immunization of female Wistar rats with Low-dose guinea pig spinal cord homogenate(GPSCH). Our experiment mainly research the aspect of thymus:(1) To observe the variation of thymic histology in different courses of experimental autoimmune encephalomyelitis (EAE) group. To observe the variation of PCNA of thymus in different courses of (EAE) group and to observe the variation of regulatory CD4+CD25+Foxp3+T cell(Treg) of thymus.(2) The a-lipoic acid was used to intervene the EAE rats, then observe the different incidence of EAE,the variation of regulatory CD4+CD25+Foxp3+T cell and the variation of PCNA of thymus.To discuss the intervening measure of a-lipoic on EAE.In order to provide new idea and theory.Methods:A total of72adult healthy female Wistar rats,were divided randomly into EAE group and ALA group,12rats for normal control group.In our experiments, EAE was induced by immunization of60rats Wistar rats with GPSCH.ALA group were injected intraperitoneal LA100mg·kg-1·d-1.This treatment was started on the first day of EAE attacked and continued for7days.Each group rats was divided into acute course,recovery course,relapse course and half-year course.To assess the EAE rats symptom by the the universal Kono score.Flow cytometry was used to analyze the expression rate of CD4+CD25+Foxp3+Treg cells in the different courses of control group, EAE group and ALA group. Immunohistochemistry was used to analyze the expression rate of PCNA in the different courses of control group,EAE group and ALA group.To observe the variation of thymic histology by light microscope.Results:1After vaccination the rats became sluggishness, lazy, anorexia, Weight loss and so on. The rats tail reduce tension and weightless, the limbs became weightless and palsy, gatism and even cannot eat by themselves after10days vaccinated. The EAE rats condition were more bad than the ALA rats. The attack frequency of the half-year EAE rats is five, the attack frequency of the half-year ALA rats is two, the morbidity of EAE is86.1%, the morbidity of ALA is50%,they have significant difference(P<0.05). The control rats was not attacked.2In acute course and relapse course of EAE and ALA group,the expression of CD4+CD25+Foxp3+Treg cells decreased significantly than that in the control group (P<0.05).In recovery course CD4+CD25+Foxp3+Treg cells raise. No significant difference was found in the expression rate of CD4+CD25+Foxp3+Treg cells between the EAE group and the ALA group.The three groups in half-year course,CD4+CD25+Foxp3+Treg cells reduced to a low level significantly,but there is no significant difference among the three groups(P>0.05).There is no Hassal's body was found in the rats thymus.3In the acute course,relapse course and half-year course of EAE rats,the cortex of thymus got thinner and the numbers of lymphocytes were reduced.The structures of thymus were loosed.We observed the increased amount of cells whose nuclear chromatins were concentrated and nuclei were pyknotic.In the relapse course the lymphocytes increased,but they were still lower than in the control group.4In acute course,recovery course and relapse course of EAE group, the expression of PCNA cells decreased significantly than that in the control group (P<0.05).No significant difference was found in the expression rate of PCNA cells between the EAE group and the ALA group(P>0.05).The three groups in half-year course PCNA cells reduced to a low level significantly,but there is no significant difference among the three groups(P>0.05).Conclusion:1The a-lipoic acid has effects in relieving the symptom of EAE and reduce the attack frequency of EAE.2CD4+CD25+Foxp3+Treg cells may play a role in the occurrence of EAE. There is significant relation between the development of EAE and CD4+CD25+Foxp3+Treg.The function of a-lipoic acid may not play through CD4+CD25+Foxp3+Treg in immune adjustment at EAE3Thymus is the sensitive immune organ,as the age added the function and the structure have both changed.The expression of CD4+CD25+Foxp3+Treg cells and PCNA were both reduced by the atrophy of thymus.It may concerned with the function of thymus.4PCNA is expressed in thymus in different courses of EAE and normal thymus.The expression of PCNA in thymus is susceptible to the environment.