The Influence Of Ndga On Expression Of Inflammatory Cytokines Of Experimental Autoimmune Encephalomyelitis In Mice

Objective: Multiple sclerosis(MS) is an immune-mediated central nervous system(CNS) disease characterized by demyelination and axonaldamage. Until now, the etiology and mechanism of MS remain unclear. Inflammation is considered to be the most important in the early stages of this disease. Cytokines play a pivotal role in the regulation of inflammatory responses and tissue repair, influencing the severity and the tendency of the disease. They orchestrate all phases of immune responses and act in highly complex, dynamic networks in a paracrine and/or autocrine fashion. In the previous study, we investigated Nrf2 activator sulforaphane could release oxidative stress and regulate immunity, providing neuroprotective effects on EAE mice. Sulforaphane is not yet in clinical practice. Instead, we searched and found another Nrf2 activator nordihydroguaiaretic acid(NDGA) which is widely used in clinic. It remains unclearly if NDGA play a role in immunoregulatory mechanism of MS and researches should be done. In this study, we observed the clinical symptoms and course in both EAE treatment group and invesgating the infuence of NDGA on IL-4, IL-6, IL-12, IL-17, IFN-γ, and TGF-β translated and expressed in EAE mice. To reveal the role of cytokines in the course of EAE and whether administration of NDGA is effective in controlling EAE.Methods: 54 female C57BL/6 mice(8 to 10 weeks old) of 18 g to 20 g body weight, were randomly divided into 3 equal groups: group EAE, group NDGA and group NDGA.Each group divided into two subgroup 10 day group and 20 day group.Then EAE mice model was established. Treatments started after onset of the first clinical signs(weaver ≥onepoint) until death, for NDGA treatment group mice receiving 10mg/kg peritoneal injection and for EAE and control group mice received 5% DEMO 10ml/kg peritoneal injection each day. Their weight and the clinical score were recorded twice a day. IL-4, IL-6, IL-12, IL-17, IFN-γ, TGF-β traslating levels in spine and spleen were detected with qrt PCR procedures, IL-4, IL-6, IL-12, IL-17, IFN-γ, TGF-β levels in brain were detected with ELISA procedures.Results:1 Compared with group EAE, the meal value of clinical score of group NDGA decreases remarkably in both 10 day and 20 day of EAE mice.2 IL-4, IL-6, IL-12, IL-17, IFN-γ, TGF-β traslating in spine and spleen:Comparing with the control group in 10 days after incident of the MS, IL-6, IL-12, IL-17, IFN-γ m RNA translation were increased in both group EAE and group NDGA, with NDGA group relatively low. However, the TGF-β, IL-4 m RNA translation in both group EAE and group NDGA were decreased, with NDGA group relatively high.Comparing with the control group in 20 days after incident of the MS, IL-6, IL-12, IL-17, IFN-γ m RNA translation were increased in both group EAE and group NDGA, with NDGA group relatively low. However, the TGF-β, IL-4 m RNA translation in both group EAE and group NDGA were decreased, with NDGA group relatively high. 3 The expression of IL-4, IL-6, IL-12, IL-17, IFN-γ and TGF-β in mice brain tissue of each group:Comparing with the control group in 10 days after incident of the MS, IL-6, IL-12, IL-17, IFN-γ m RNA translation were increased in both group EAE and group NDGA, with NDGA group relatively low. However, the TGF-β, IL-4 m RNA translation in both group EAE and group NDGA were decreased, with NDGA group relatively high.Comparing with the control group in 20 days after incident of the MS IL-17, IFN-γ were increased in both group EAE and group NDGA, with NDGA group relatively low.TGF- β in NDGA group relativelyhigher than EAE group.Conclusion:1 The onset and alleviation of EAE are associated with the dynamic change of cytokines.2 NDGA could reduce the neural function damage severity of the disease of EAE mice, promote nerve functional recovery, and has a protective effect on experimental autoimmune myelitis mice.3 NDGA could lower the expression of proinflammatory cytokines(IL-6, IL-12, IL-17, IFN- γ) in the EAE brain, spinal cord and spleen tissue and increase the expression of the anti-inflammatory cytokine IL-4 and TGF-β, with anti-inflammatory effects.4 NDGA plays a nerve protective role through controlling immune responses and the maintenance of immune balance.