Therapeutic Effect Of Tripterygium Wilfordii Polyglycosidium On Experiment Autoimmune Encephalomyelitis In Rats

Objective To establish the animal model of experimental autoimmune encephalomyelitis(EAE) and observe the expression of caspase-3 and IFN-γin spinal cord of EAE rats, explore the possible protective function mechanism of Tripterygium wilfordii Ployglycosidium (TWP) to EAE rats, seek for the efficient path of multiple sclerosis's treatment .Methods Sixty female Wistar rats were randomly divided into four groups: normal control group (CON), EAE mode group (EAE), TWP1 group, TWP2 group. The EAE animal model was established in rats by immunizing rats with guinea pig spinal cord homogenate (GPSCH) and complete freund's adjuvant(CFA). The CON group rats were sacrificed in the 21st day. The animals of other groups were divided into three subgroups :the 7th day sacrificed ,the 14th sacrificed and the 21st sacrificed. The Rats were fed with TWP(10mg/kg.d) in TWP1 group, TWP (30mg/kg.d) in TWP2 group , starting from the 1st day after immunization to the day rats were Sacrificed. Nerve function scores of rats were examined daily. Histological examinations were performed on the sections of spinal cord with the aid of hematoxylin- eosin staining. The number of inflammation in the central nervous system (CNS) was counted under the optical microscope .And the expressions of caspase-3 and IFN-γin spinal cord were detected by using immunohistochemistry technique.Results1.Incidence of a disease: No ill rats in CON group .The incidence of EAE group was 86.67%. The incidence of TWP1 and TWP2 groups was lower than that of the EAE group (P <0.05), the incidence of TWP2 group was lower comparing with the TWP1 group (P<0.05). 2.Ethology appraisal:The neurological function score of TWP1 and TWP2 groups was lower than that of the EAE group at the 14th day and the 21th day (P <0.05), and that of the 14th day of TWP2 group was lower comparing with the TWP1 group(P <0.05). Comparing the three time points in each group, the 7th day of group, the 21st day of group compared with the 14th day of group was significantly different (P <0.05); the 7th day and the 21st day of group, no significant difference between groups (P > 0.05).3.Histopathological findings: There was no inflammatory cell infiltration of the CON group, but the inflammatory cell was found in the spinal cord of the each stage group of EAE group and TWP1,TWP2 group. The nurnber of inflammatory cell of TWP1,TWP2 group was fewer than that of the EAE group at the 14th day and the 21th day (P <0.05),and that of the TWP2 group was lower than the TWP1 group (P <0.05). Comparing the three time points in each group, the 7th day of group, the 21st day of group compared with the 14th day of group was significantly different (P <0.05); the 7th day and the 21st day of group, no significant difference between groups (P > 0.05).4.Caspase-3 and IFN-γexpression: There was no caspase-3 and IFN-γcells in spinal cord of the CON group; but the caspase-3 and IFN-γcell could be seen in the spinal cord of the EAE group and TWP1,TWP2 group,and the number of caspase-3 and IFN-γcell of TWP1,TWP2 group was fewer than that of the EAE group (P <0.05),that of the 14th day of TWP2 group was lower than the TWP1 group (P <0.05). Comparing the three time points in each group, the 7th day of group, the 21st day of group compared with the 14th day of group was significantly different (P <0.05); the 7th day and the 21st day of group, no significant difference between groups (P > 0.05).ConclusionTWP intervention has a protective effect on EAE in rats, reduce the incidence of EAE rats and showed dose-dependent, which may reduce the EAE rat spinal cord with caspase-3 and IFN-γexpression.