摘要
目的探讨载脂蛋白E（ApoE）对实验性自身免疫性脑脊髓炎（EAE）小鼠发病前期脾脏巨噬细胞分泌基质金属蛋白酶-9（MMP-9）、基质金属蛋白酶组织抑制因子-1（TIMP—1）及炎症因子的影响。方法按照随机数字表法，将20只ApoE基因敲除型（ApoE-/-）C57BL／6J小鼠分为ApoE-／- EAE组和ApoE-/-对照组，20只野生型（WT）C57BL／6J小鼠分为WTEAE组和WT对照组，每组10只。采用髓鞘少突胶质细胞糖蛋白35．55（MOG35—55）作为抗原诱导ApoE-/- EAE组和WTEAE组小鼠制成EAE模型，ApoE-/-对照组和WT对照组小鼠以生理盐水代替。提取造模后7d时小鼠脾脏巨噬细胞进行体外培养，ApoE-/- EAE组和WTEAE组巨噬细胞以MOG35—55刺激，ApoE-/-对照组和WT对照组巨噬细胞予等量生理盐水处理，48h后采用酶联免疫吸附法（ELISA）检测细胞培养液中MMP-9、TIMP-1、干扰素-γ（IFN-γ）、肿瘤坏死因子-α（TNF-α）及白介素-6（IL-6）含量。结果WTEAE组巨噬细胞的MMP-9、IFN-γ、TNF-α及IL-6分泌量均明显高于WT对照组，ApoE-／-EAE组巨噬细胞的MMP-9、IFN—γ、TNF-α及IL-6分泌量均明显高于ApoE-／-对照组，ApoE-／-EAE组巨噬细胞的MMP-9、IFN-γ、TNF-α及IL-6分泌量均明显高于WTEAE组，WTEAE组巨噬细胞的TIMP-1分泌量明显高于WT对照组，差异均有统计学意义（P〈0．05）。结论在EAE发病前期，脾脏巨噬细胞分泌的MMP-9及炎症因子增多，而ApoE表达的缺乏进一步促进巨噬细胞分泌更多的MMP-9及炎症因子，故给予ApoE补充可能会通过调节巨噬细胞分泌MMP-9及炎症因子．起到保护血脑屏障完整性、减轻炎症反应的作用。
Objective To explore the effect of apolipoprotein E （ApoE） on secretions of matrix metalloproteinase-9 （MMP-9）, matrix metalloproteinase tissue inhibitor-1 （TIMP-1） and inflammatory factors in spleen maerophages at early stage of experimental autoimmune encephalomyelitis （EAE）. Methods In accordance with random number table, 20 ApoE knockout （ApoE-/-） C57BL/6J mice were divided into ApoE-/- EAE group and ApoE-/- control group （n=10）; and 20 wild type （WT） C57BL/6J mice were divided into WT EAE group and WT control group （n=10）. Myelin oligodendrocyte glycoprotein 35-55 （MOG35-55） was used as antigen to induce EAE models in the ApoE-/- EAE group and WT EAE group, and MOG35-55 was replaced by normal saline in ApoE-/- control group and WT control group. The spleen macrophages were isolated from mice after 7^th d of modeling and cultured in vitro; the macrophages of the ApoE-/- EAE group and WT EAE group were stimulated by MOG35-55, while the macrophages of the ApoE-/- control group and WT control group were treated with equivalent normal saline. Forty-eight h after that, the contents of MMP-9, TIMP-1, interferon gamma-γ（IFN-γ）, tumor necrosis factor-α（TNF-α） and interleukin-6 （IL-6） in the culture medium were determined by ELISA. Results The contents of MMP-9, IFN-γ, TNF-α and IL-6 in the WT EAE group were significantly higher than those in the WT control group （P〈0.05）; the contents ofMMP-9, IFN-γ,TNF-α and IL-6 in the ApoE-/- EAE group were significantly higher than those in the ApoE-/- control group （P〈0.05）; the content of MMP-9, IFN-γ, TNF-α and IL-6 in ApoE-/- EAE group were significantly higher than those in the WT EAE group （P〈0.05）. The TIMP-1 secretion ofmacrophages in WT EAE group was significantly higher than that in WT control group （P〈0.05）. Conclusions At early stage of EAE, macrophages secrete high levels of MMP-9 and inflammatory factors, and the deficiency of ApoE further contributes to the secretions of MMP-9 and inf
作者
卢珍梅
卢文玉
唐玉兰
范丽君
李一锋
韦俊杰
韦云飞
Lu Zhenmei;Lu Wenyu;Tang Yulan;Fang Lijun;Li Yifeng;Wei Junjie;Wei Yunfei(Departraent of Neurology, First Affiliated Hospital, Guangxi Medical University, Nanning 530021, China;Department of Neurology, Guangxi Zhuang Autonomous Region People＇s Hospital, Nanning 530021, China;Department of Neurology, Second Affiliated Hospital, Guangxi Medical University, Nanning 530007, China)
出处
《中华神经医学杂志》
CAS
CSCD
北大核心
2018年第2期136-141,共6页
Chinese Journal of Neuromedicine
基金
（1）国家自然科学基金（81460194、81260188）（2）广西自然科学基金（2015GXNSFAA139212、2012GXNSFAA053082）
