| Objective:To observe the effects of oxymatrine(Matrine, MAT) on experimental autoimmune encephalomyelitis(experimental autoimmune encephalomyelitis, EAE) of the clinical symptoms and pathological histology in rats, rats in the pathogenesis of amyloid beta(beta-Amyloid, A), beta secretase(B-site APP cleaving enzyme 1 BACE-1, myelin basic protein(Myelin), basic protein, MBP), S100 B protein(S100B protein, S100B), neuron specific enolase(neuron specificenolase, NSE), brain derived neurotrophic factor(brain derived, neurotrophic factor, BDNF) expression changes of Matrine on EAE axonal control effect and mechanism of injury, intended for the multiple sclerosis drug new ideas. Methods:1 The establishment of EAE model: under sterile conditions, the guinea pig spinal cord homogenate(guinea pig spinal cord homogenate, GPSCH), and complete Freund’s adjuvant with BCG(6mg/ml complete Freund adjuvant, CFA) mixed in equal volumes, repeatedly beat preparation stability of the emulsion as antigen, 6 ~ 8 week old female Wistar rats footpad injection of antigen emulsion induced EAE model. Self immunity date, observe and record the changes of body weight, the rats activity changes, clinical symptoms and nerve function score.2 Animal groups and the treatment of EAE:30 Wistar female rats were randomly divided into three groups: after immunization with EAE group(EAE), MAT group(MAT), methylprednisolone group(MET), 10 rats in each group. At the same time 10 healthy rats were used as control group(Normal). Immune from that day, four groups of rats were given different interventions, group MAT given daily doses of 1 3.3ml/kg(200mg/kg) Marine Injection intraperitoneal injection, group MET given daily doses of 1 3.3ml/kg(1mg/kg) and dexamethasone injection by intraperitoneal injection, normal control group and model group with the same volume of saline instead of treatment, four groups of the rats were 16 days of continuous intervention.3 Sample collection: the immune seventeenth days, using 10% chloral hydrate anesthesia, the orbital venous plexus of blood 2ml, serum pre cooling system; saline perfusion after complete, the rats were left for specimens, paraffin section, the right brain immediately put into liquid nitrogen retained backup.4 Related indexes: the hematoxylin and eosin(HE) staining and Pierre Su Minsky’s(Bielschowsky staining) staining was used to observe the inflammatory cell infiltration and demyelination, analysis of the expression of A beta immunohistochemistry, enzyme-linked immunosorbent assay(ELISA method) to detect MBP, S100 B, NSE, reverse transcription polymerase chain reaction(RT-PCR analysis of BACE-1 and BDNF content).Results:1 EAE incidence:with 10 animals in each group, the rats in normal group were not observed the incidence, the incidence of all 10 rats in model group, MAT group, MAT group of 6 patients, 4 patients, group MET with 2 patients. Fisher exact analysis results showed: the normal control group the incidence was significantly lower than that of model group(P < 0.05); group MAT, the incidence rate of MET group rats was lower than that of model group, but the difference was not statistically significant(P > 0.05).2 Body weight trend:Rats in the normal group from the second day average weight increase immunity, the remaining three groups of average weight showed a downward trend after rising. The weight of MAT group was significantly lower than that of model group decreased significantly(P < 0.05), also lower than those in MET group(P < 0.05).3 Clinical score trend:I The normal group of nerve in this study were maintained for 0. Immune seventeenth days, the average neurological function in the rats of model group was significantly higher than the scores of two drug intervention group(P < 0.05). There was no significant difference between the two drug intervention group the average neurological function score(P > 0.05).4 CNS histopathology : The average score of model group, inflammatory infiltration and demyelination were higher than those in control group, the difference was significant(P < 0.05). The average score of MAT group, inflammatory infiltration and demyelination were significantly lower than those in the model group(P < 0.05). Compared with MAT group, MET group, the average score of inflammatory infiltration was significantly lower(P < 0.01), there was no significant difference between the scores of the myelin sheath(P > 0.05).5 BACE-1 and BDNF m RNA expression in cervical spinal cords:BACE-1 in the brain, the expression of BDNF m RNA: compared with the normal control group, BDNF model group, m RNA content decreased significantly(P < 0.05). The expression of BDNF m RNA in MAT group were significantly higher than those in the model group(P < 0.05), MAT group and MET group in BACE-1, BDNF m RNA expression had no significant difference(P > 0.05).6 Aβ content in the brain:The expression of Aβ: compared with the normal control group, model group, Aβ content increased significantly(P < 0.05); two of the drug intervention group were lower than model group, the expression of Aβdecreased significantly(P < 0.05), the differences between groups was significant(P < 0.05).7 MBP, S100 B, NSE in Serum: compared with the normal control group, model group rats serum MBP, S100 B, NSE were significantly increased(P < 0.05). MAT group MBP, S100 B, NSE levels were significantly lower than those in the model group(P < 0.05), MET group in the inhibition of S100 B were increased than group MAT, the difference was statistically significant(P < 0.05). Conclusion: MAT has good preventive and therapeutic effect on Wistar rats EAE. |
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