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目的观察二黄胶囊对实验性自身免疫性脑脊髓炎小鼠(EAE)细胞免疫的影响。方法皮下注射髓鞘少突胶质细胞糖蛋白35-55(MOG35-55)与完全弗氏佐剂(CFA)混合的抗原,辅以腹腔注射百日咳毒素(PTX),建立EAE模型。造模后每天记录各组小鼠体质量及神经功能评分;用流式细胞仪检测小鼠外周血T淋巴细胞亚群及自然杀伤(NK)细胞;采用HE染色进行脑组织病理学观察。结果模型组小鼠造模后第9天陆续发病,第22天达到高峰,发病急性期(造模后第25天)T淋巴细胞亚群CD3+、CD4+明显比正常组增高(P<0.01),CD4/CD8比值也有所升高(P<0.05);发病缓解期(造模后第40天)T淋巴细胞亚群CD8+和NK细胞明显下降(P<0.05);病理检查发现EAE小鼠脑组织神经元细胞核固缩,小静脉周围炎性细胞浸润形成袖套样改变。二黄胶囊治疗后,EAE小鼠神经功能评分比模型组明显降低(P<0.05),脑组织炎性浸润程度减轻,发病急性期外周血T淋巴细胞亚群CD4+/CD8+比值显著降低(P<0.01);恢复期T淋巴细胞亚群(CD3+、CD8+)及NK细胞明显增加(P<0.05)。结论二黄胶囊可在EAE小鼠发病的不同时期通过调节T淋巴细胞亚群(CD3+,CD4+,CD8+,CD4+/CD8+)及NK细胞而起到防治EAE作用。
Objective To observe the effect of Erhuang Capsule on cellular immunity of experimental autoimmune encephalomyelitis (EAE) mice. Methods EAE model was established by subcutaneous injection of myelin oligodendrocyte glycoprotein 35-55 (MOG35-55) and complete Freund’s adjuvant (CFA) mixed antigen, supplemented by intraperitoneal injection of pertussis toxin (PTX). The body weight and neurological function scores of each group were recorded every day after modeling; peripheral blood T lymphocyte subsets and natural killer (NK) cells were detected by flow cytometry; brain pathology was observed by HE staining. Results The mice in the model group developed on the 9th day after modeling and peaked on the 22nd day. In the acute phase (25th day after modeling), the T lymphocyte subsets CD3+ and CD4+ were significantly higher than those in the normal group (P<0.01). CD4/CD8 ratio also increased (P<0.05); CD8+ and NK cells in T lymphocyte subsets decreased significantly at the onset of remission (40 days after modeling) (P<0.05); pathological examination found EAE mouse brain tissue Neuronal nucleus pyknosis, infiltration of inflammatory cells around the small veins formed cuff-like changes. After treatment with Erhuang Capsule, the neurological score of EAE mice was significantly lower than that of the model group (P<0.05). The degree of inflammatory infiltration in the brain tissue was reduced, and the ratio of CD4+/CD8+ in peripheral blood T lymphocyte subsets was significantly decreased in the acute phase of disease (P< 0.01); Recovery of T lymphocyte subsets (CD3+, CD8+) and NK cells increased significantly (P<0.05). Conclusion Erhuang capsule can prevent and treat EAE by regulating T lymphocyte subsets (CD3+, CD4+, CD8+, CD4+/CD8+) and NK cells at different stages of EAE mouse onset.