二黄胶囊对实验性自身免疫性脑脊髓炎小鼠细胞免疫的影响

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。

Effects of Erhuang Capsule on Cell Immunity of Mice with Experimental Autoimmune Encephalomyelitis

Objective To observe the effects of Erhuang Capsule on cell immunity of mice with experimental autoimmune encephalomyelitis(EAE). Methods EAE models C57BL/6 mice were induced by injecting subcutaneously myelin oligodendrocyte glycoprotein 35-55 peptide in complete Freund's adjuvant(CFA), assisted by injecting intraperitoneally with pertussis toxin (PTX) . The animals were weighted and graded daily for clinical disability. At 12, 25 and 40th day after immunization, lymphocyte subgroups and NK cells in the peripheral blood of mice were detected with flow cytometry. Pathologic identification of the brain tissue were done by HE staining. Results The mice in model group were affected beginning on day 9 and reached the peak on day 22. The CD3⁺ and CD4⁺ in the lymphocyte subgroups increased significantly(P<0.01), and the ratio of CD4/CD8 of T lymphocyte subsets elevated in active phase(P<0.05). At the remission stage, the percentage of T lymphocyte subset CD8⁺ and NK cells declined significantly(P<0.05). In pathology, infiltrated inflammatory cells were observed to form sleeves around the vein and the cerebral neurons showed nucleus pyknosis. After the treatment of Erhuang Capsule, the neurological score declined(P<0.05) and HE staining showed mild inflammatory infiltration. As compared with the model group, the ratio of CD4/CD8 of T lymphocyte subsets was significantly lowered in active phase (P<0.01), while, the levels of T lymphocyte subset(CD3⁺ and CD8⁺) and NK cells increased obviously at the remission stage(P<0.05). Conclusion Erhuang Capsule may prevent and cure EAE by regulating the T-lymphocyte subsets(CD3⁺, CD4⁺, CD8⁺ and CD4⁺/CD8⁺ ratio) and NK cells.