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目的观察冬凌草甲素(ORI)对实验性自身免疫性脑脊髓炎(EAE)的预防作用,并探讨其可能机制。方法 EAE小鼠发病前给予ORI治疗,以PBS为阴性对照,观察各组小鼠发病情况。通过对EAE小鼠临床评分和脊髓病灶病理学检测判断ORI的预防效果;通过5,6-羧基荧光素二乙酸盐琥珀酰亚胺酯(CFSE)标记检测CD4+T细胞增殖,流式细胞术检测白细胞介素17(IL-17)阳性细胞占CD4+T细胞的百分比、γ干扰素(IFN-γ)阳性细胞占CD4+T细胞的百分比,ELISA检测IL-1β水平。结果 ORI能显著改善EAE的临床评分、减少脊髓病变部位炎性细胞浸润;ORI能抑制EAE小鼠髓鞘少突胶质细胞糖蛋白(MOG)反应性T细胞增殖;与PBS组相比,ORI组IFN-γ+CD4+T细胞和IL-17+CD4+T细胞减少;ORI抑制脾脏肿大、淋巴细胞增殖和IL-1β分泌。结论 ORI能减轻EAE严重程度,与减少IL-1β水平、抑制T细胞增殖及辅助T细胞分化有关。
Objective To observe the preventive effect of oridonin (ORI) on experimental autoimmune encephalomyelitis (EAE) and to explore its possible mechanism. Methods EAE mice were given ORI before onset, PBS was used as negative control to observe the incidence of mice in each group. The preventive effect of ORI was judged by the clinical score of EAE mice and the pathological examination of spinal lesions. The proliferation of CD4 + T cells was detected by CFSE labeling. Flow cytometry The percentage of IL-17 positive cells to CD4 + T cells, the percentage of interferon-γ (IFN-γ) positive cells to CD4 + T cells were detected by ELISA. The level of IL-1β was detected by ELISA. Results ORI could significantly improve the clinical scores of EAE and reduce inflammatory cell infiltration in spinal cord lesion. ORI can inhibit the proliferation of myelin oligodendrocyte glycoprotein (MOG) reactive T cells in EAE mice. Compared with PBS group, ORI IFN-γ + CD4 + T cells and IL-17 + CD4 + T cells decreased; ORI inhibited spleen enlargement, lymphocyte proliferation and IL-1β secretion. Conclusion ORI can reduce the severity of EAE, and reduce the level of IL-1β, inhibit T cell proliferation and assist T cell differentiation.