| Objective:Sinomenine,a alkaloid monomer extracting from a plant Sinomenium acutum,has been used to relieve rheumatoid arthritis in human having well therapeutic effect and a few side effect.Recent studies have shown that sinomenine possesses favourable immunosuppressive properties. Whether this reagent is effective in human multiple sclerosis(MS),an inflammatory demyelination in the CNS,is not known.Research report has been obtained that sinomenine suppress the activity of proinflammatory nuclear factor KappaB(NF-κB)in the brain to ameliorate experimental autoimmune encephalomyelitis(EAE),an animal model of MS.We thus further investigated the therapeutic effect and mechanism of Sinomenine on EAE through the orientation of regulatory cytokine interleukin-10(IL-10) and transforming growth factor-β1(TGF-β1).Methods:The animal model was established in female Wistar rats by immunizing rats with guinea pig spinal cord homogenate(GP-SCH), complete freund's adguvant(CFA)and pertussis vaccine(PV).Fifty wistar rats were randomly divided into four groups:five rats in normal control group,each fifteen rats in EAE group,Sinomenine group and Prednisone group,then the later three group were randomly divided into three subgroups,each subgroup have five rats.Rats were fed with sinomenine (100mg/kg.d)in sinomenine group,while prednisone group were fed with prednisone(5mg/kg.d),starting from the 1st day after immunization to the day rats were sacrificed.The animals of control group were sacrificed at the 15th day.The three subgroups of other groups were sacrificed at the 11th day, the 15th day and the 21st day.The severity of EAE was scored on a scale 0～5 according to the signs and symptoms.Histological examinations were performed on the sections of brain and 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 IL-10 and TGF-β1 in brain tissue were detected by using immunohistochemistry technique and optical density detection.Experimental results were analyzed with SPSS13.0.Results:1.No rats in normal control group have disease.Comparing to EAE group,the EAE of Sinomenine and Prednisone group's rats have prolonged eclipse period(p＜0.05),degraded morbility,weight loss and neurological deficit scores(p＜0.05).Comparing to Prednisone group,the EAE of Sinomenine group's eclipse period,morbility,weight loss and neurological deficit scores have no difference(p＞0.05)2.The CNS tissue's hematoxylin-eosin(H-E)staining histopathology results of EAE group's rats indicate that CNS already have inflammation focus at the 11th day,the extent and amount of the inflammation focus increased at the 15th day,then decreased at the 21th day.Comparing to the 15th day subgroup of EAE group,the same subgroup of Sinomenine and Prednisone group have decreased CNS inflammation focus(p＜0.05). Comparing to the 15th day subgroup of Prednisone group,the same subgroup of Sinomenine's CNS inflammation focus have no difference(p＞0.05)3.Immunohistochemistry and image analysis results of each group rats indicate that the CNS expression level of IL-10 and TGF-β1 has no difference in the 11th day subgroup(p＞0.05)and up-regulated in the other subgroup(p＜0.05)comparing with normal control group.With the prolonged course of disease,the 15th day subgroup comparing with the 11th day,the 21th day subgroup comparing with the 15th day have higher level of IL-10 and TGF-β1in the CNS(p＜0.05).Comparing to the same subgroup of the EAE group,Sinomenine and Prednisone group's rats have higher level of IL-10 and TGF-β1in the CNS(p＜0.05).Comparing to the subgroup of Prednisone group,the same subgroup of Sinomenine's CNS level of IL-10 and TGF-β1have no difference(p＞0.05)Conclusion:1.Sinomenine have protect effect on EAE rats as sinomenine could decrease EAE morbility,prolong eclipse period,ameliorate clinical manifestation and degrade neurological deficit scores.2.One of the mechanisms related to sinomenine's protection on EAE is up-regulation the level of IL-10 and TGFβ1 in the CNS.
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