| Objective:In the study, we established a model that uses different dose fingolimod and prednisone to intervene experimental autoimmune encephalomyelitis (EAE) of the Wistar rats intending to find the difference of two measurements, as well as observe the changes on clinical, pathological and expression level of Bax & TRL9 of brain and spine tissue during EVE rats treated by the fingolimod, so that can estimate that potential mechanism which fingolimod effects on EVE could be Multiple Sclerosis(MS). The finding may make good foundation for new medications.Methods:The animal model was established in female Wistar rats by immunizing rats with guinea pig spinal cord homogenate(GPSCH), complete freund's adguvant(CFA) and pertussis vaccine (PV).Seventy wistar rats were randomly divided into seven groups:ten rats in normal control group, ten rats in EAE group, different doses of FTY720 group and prednisone group. Rats were fed with FTY720 in different doses FTY720 group, while prednisone group were fed with prednisone, starting from the day rats were injected by antigen to the day rats were put to death. All of rats were executed at the 18th day after immunization, and the scores of symptom and changes of body weight were recorded in the period. Brain and spinal cords were extracted for pathology observation with the aid of hematoxylin-eosin and Luxol Fast Blue myelin staining.Count the number of inflammation on the central nervous system (CNS) under optical microscope. We find that the expressions of BAX and TLR9 in brain and spine tissue and Spinal cord white quality by using immunohistochemistry technique. SPSS17.0.were use to analyzed Experimental resultsResults:1. Observation on clinical manifestations:(1) Rats in normal group keep normal. (2) Compared with EVE group, rats in low dose group have no specific changes in morbidity and eclipse period (p>0.05) while they show lower nerves function mark and weight losing (p<0.05). and rats in middle dose group, higher dose group, high dose group and Prednisone group can be seen improved obviously on the indicators of eclipse period,morbidity, weight and nerves function mark (p>0.05).2. Observation on pathological results:(1) inflammation focus:①The brain tissue's hematoxylin-eosin(HE) staining histopathology results of EAE group's rats indicate that CNS already have inflammation while no change in normal control group.②Comparing with EAE group, the extent and amount of the inflammation focus decreased in the different doses FTY720 and Prednisone group(p<0.05).③Comparing with the Prednisone group, the 0.1 and 0.3mg/kg-d groups'CNS inflammation focus have no difference(p>0.05).④There are few inflammation focus in the 1.0mg/kg·d group,and have no difference between the normal control group. (2) Luxol fast bule-Cresyl fast violet myelin staining indicated no demyelination in normal group, while typical demyelination lesions were serious occurred in EAE group.3. Observation on results of Immunohistochemistry:(1) Compared to normal group, the expression level of Bax in EVE group falls down(P<0.05), while their expression level of TLR9 raises (P<0.05). (2) Comparing with EVE group,expression of TLR9 in different dose of FTY720 group and prednisone group have raised, while expression level of Bax dropped (p<0.05), and also find that middle dose group, higher dose group and prednisone group have no significant differences(p>0.05),while the difference between these three groups and high dose group was statistical significantly(p<0.05).Conclusion:1. FTY720 has amelioration effect on EAE rats and the effects were in dose-dependent fashion.2. one of the mechanism which FTY720 play its amelioration effect could be making expression level of TLR9 in CNS down-regulation while up-regulation the level of BAX. |
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