Preliminary Study On Anti-inflammatory Effects Of Grape Seed Proanthocyanidins On EAE

Objective:In this study,we try to explore the anti-inflammatory effect of grape seed proanthocyanidins?GSE?on lipopolysaccharide?LPS?-stimulated BV-2 microglia?MG?,and further explore the treatment effect of oral GSE in C57BL/6 mice with the experimental autoimmune encephalomyelitis?EAE?models induced by myelin oligodendrocyte glycoprotein peptides 35-55?MOG35-55?,providing experimental basis for further clinical treatment of multiple sclerosis?MS?.Methods:A:The anti-inflammatory effect of GSE on BV-2MG?1?The effect of different concentrations of GSE on proliferation of BV-2MG was examined by MTT assay;?2?BV-2MG were divided into blank group,LPS control group,LPS+GSE group.The levels of inflammatory factors were detected by ELISA;?3?The morphological changes of BV-2MG were observed by HE;?4?The NF-?B phosphorylation in BV-2MG were detected by Western blot.B:The therapeutic effect of GSE on EAEThe experimental animals were randomly divided into GSE treatment group and EAE control group.The EAE model was induced by MOG_35-55 peptide.GSE and saline were respectively administered by gavage every day from the third day to the 28th day after immunization.?1?Clinical score was evaluated every other day;?2?The animals were sacrificed at day 28 after immunization.Spinal cords were collected for HE staining and Luxol Fast Blue staining;?3?The levels of inflammatory factors in spleen cells were detected by ELISA.Results:A:?1?MTT results showed that:GSE did not affect the survival of normal cultured BV-2MG in the concentration range of 100mg·L^-1;?2?ELISA results showed that:Compared with LPS group,GSE could inhibite the release of inflammatory in LPS-stimulated BV-2MG;?3?HE staining showed that:Compared with LPS group,GSE could reverse the morphological changes of BV-2MG cells stimulated by LPS;?4?Western blot results showed that:Compared with the LPS group,GSE was able to reduce the phosphorylation of p65NF-?B in BV-2MG.B:Compared with EAE control group:?1?GSE can improve EAE symptoms;?2?HE staining and Luxol Fast Blue staining results show that:GSE can improve the inflammatory cell infiltration of the spinal cords and reduce white matter demyelination;?3?ELISA results showed that:GSE can inhibit the secretion of inflammatory cytokines in splenocytes.Conclusion:GSE has a good effect on EAE treatment.The therapeutic effect may be to reduce the inflammatory cell infiltration and myelin sheath loss of the CNS,reduce the NF-?B phosphorylation of inflammatory cells and secretion of inflammatory factors,thereby improving the CNS inflammation microenvironment,and achieving the prevention in the occurrence and development of EAE.