Cornuside ameliorated experimental autoimmune encephalomyelitis by limiting the recruitment of CD4+ T lymphocytes in the spinal cord

Abstract We conducted this study to determine whether cornuside could improve the neurological deficit symptoms of experimental autoimmune encephalomyelitis (EAE) rats, as well as determine the potential involvement of CD4+ T lymphocytes, vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), and tumor necrosis factor-α (TNF-α). Altogether, 32 Lewis rats were randomly divided into control, EAE, EAE/prednisolone, and EAE/cornuside, wherein their neurological function was assessed every day. CD4+ T lymphocyte recruitment into the spinal cord (SC) was evaluated using immunohistochemistry. The VCAM-1, ICAM-1 and TNF-α mRNA expressions in the SC were determined by real-time quantitative PCR, and the VCAM-1 and ICAM-1 proteins were determined by western blotting. Compared to the control group, the EAE group rats with neurological deficits had enhanced CD4+ T lymphocyte infiltration and higher expression levels of VCAM-1, ICAM-1, and TNF-α in the SC. Meanwhile, compared with the EAE group, the EAE/cornuside and EAE/prednisolone groups had lower neurological scores, less CD4+ T lymphocyte infiltrations, and lower expression levels of VCAM-1, ICAM-1, and TNF-α in the SC. Thus, cornuside ameliorated EAE, which could be owed to the inhibition of CD4+ T lymphocyte recruitment and VCAM-1, ICAM-1, and TNF-α expressions in the SC.

摘要 本研究旨在探究山茱萸苷（cornuside）能否改善实验性自身免疫性脑脊髓炎（experimental autoimmune encephalomyelitis, EAE）模型大鼠的神经功能缺损症状，并明确CD4+ T淋巴细胞、血管细胞黏附分子-1（vascular cell adhesion molecule-1, VCAM-1）、细胞间黏附分子-1（intercellular adhesion molecule-1, ICAM-1）及肿瘤坏死因子-α（tumor necrosis factor-α, TNF-α）在其中的潜在介导作用。
共纳入32只Lewis大鼠，随机分为对照组、EAE模型组、EAE/泼尼松龙组及EAE/山茱萸苷组，每日评估大鼠神经功能。采用免疫组织化学法检测脊髓（spinal cord, SC）中CD4+ T淋巴细胞的募集情况；通过实时定量聚合酶链反应检测脊髓中VCAM-1、ICAM-1及TNF-α的mRNA表达水平；采用蛋白质印迹法检测脊髓中VCAM-1与ICAM-1的蛋白表达水平。
与对照组相比，EAE模型组大鼠出现神经功能缺损，脊髓内CD4+ T淋巴细胞浸润增强，VCAM-1、ICAM-1及TNF-α的表达水平显著升高。与EAE模型组相比，EAE/山茱萸苷组与EAE/泼尼松龙组的神经功能评分更低，脊髓内CD4+ T淋巴细胞浸润程度更轻，且VCAM-1、ICAM-1及TNF-α的表达水平均显著降低。
综上，山茱萸苷可有效缓解实验性自身免疫性脑脊髓炎，其作用机制可能与抑制脊髓内CD4+ T淋巴细胞募集，以及下调VCAM-1、ICAM-1和TNF-α的表达密切相关。