| Multiple sclerosis (MS) is a CNS autoimmune demyelination. The mechanisms of MS was not very clear, but it is generally believed that T cells play a key role. The disproportion of Th1/Th2 and disorder of their cytokines is the main cause of MS. Thl cells produce inflammatory cytokins, including IL-2, IFN-y, TNF-a and TNF-βwhich initiate and aggravate the MS myelin damage; while Th2 cells secrete anti-inflammatory cytokines IL-4, IL-5, IL-6, IL-10 and IL-13 which supress the autoimmune reaction. Therefore, decrease the proportion of Thl/Th2 and restore the balance of Thl/Th2 is an effective measure to control autoimmunity inflammation and alleviate MS.At present, treatment of MS in domestic and international is the uses of cortical hormone, immunity inhibitor etc., but all of them have weak therapeutic effects, serious side effects and high price. In recent years the Chinese medicine was used to treat MS and some progress has been made. It has been found that some Chinese medicine with anti-inflammation, immunity regulation, anti-oxidation and neural protection have inhibitory effect on inflammation and protective function on CNS. Radix Angelicae Biserratae Doubleteeth Angelica Root (Duhuo) is the herb of dispelling wind and eliminating dampness. Modern pharmacology research has indicated it's functions of anti-inflammation, immune regulation, anti-tumor, anti-bacterial, etc. In recent years Duhuo has also been used to treat Alzheimer's disease experimentally by inflammatory suppression, immunity increase and anti-apoptosis. So we suppose that Duhuo may also intervenes the autoimmune mechanism of MS, thereby reduces CNS damage and alleviate EAE disease process.Experimental allergic encephalomyelitis (EAE) is recognized internationally as an ideal animals model of MS. It has similar characterization with human MS in clinical, biochemical, immunological and pathological features. In this study we firstly use EAE model to detect the immunological mechanism of Duhuo on MS/EAE, aim to give further experimental data for the clinical use of Duhuo on human MS.Methods and results:1. The establish of EAE model and grade of CNS functionMOG35.55 immunized C57BL/6 mice was administered in stomach with different dose of Duhuo for 40 days, and was graded every day according to CNS function test standards. The results showed that high dose of Duhuo (36 mg/kg) can improve the EAE CNS function, decrease clinical score obviously (P<0.05 vs EAE model group)2. Lymph cells proliferation assayMOG35-55 immunized C57BL/6 mice was administered in stomach with different dose of Duhuo for 14 days. Spleen lymph cells was collected and stimulated by 10μg/ml MOG35-55 for 3days, then the cells were harvested for proliferation assay with MTT. The results showed that high dose of Duhuo inhibited the proliferation obviously (P<0.05 vs EAE model group)。3. Lymph cells releasing cytokine assayMOG35-55 immunized C57BL/6 mice was administered in stomach with different dose of Duhuo for 14 days. Spleen lymph cells was collected and stimulated with 10μg/ml MOG35-55.for 3days, then the supernentant was harvested for detection of Thl cytokine IFN-y and Th2 cytokine IL-13 by Elisa assay. The results showed that high dose of Duhuo decreased IFN-y and increased IL-13 obviously (P<0.05 vs EAE model group), indicating the down regulation of proportion of Th1/Th2 and restoration of the balance of Th1/Th2, resulting in the alleviating of MS/EAE.Conclusion:(1). Duhuo improved EAE CNS founction and decrease clinical score obviously;(2). Duhuo inhibited Lymph cells proliferation obviously;(3).Duhuo decreased IFN-γand increased IL-13 obviously (P<0.05 vs EAE model group), indicating the down regulation of proportion of Th1/Th2 and restoration of the balance of Th1/Th2, resulting in the alleviating of MS/EAE. |
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