载脂蛋白E对实验性自身免疫脑脊髓炎IL．17、IL-6、TNF-α表达的影响  

Effect of apolipoprotein E on inflammation in mice with experimental autoimmune encephalomyelitis

作　　者：雷茵[1] 韦俊杰[2] 唐玉兰[1] 杨程程[1] 郑明华[1] 赵伟佳[1] 韦云飞[1] 陶敏[3] 

机构地区：[1]广西医科大学第一附属医院神经内科,南宁530021 [2]广西壮族自治区人民医院神经内科,南宁530021 [3]玉林市第一人民医院神经内科,537000

出　　处：《中华神经医学杂志》2016年第5期433-438,共6页Chinese Journal of Neuromedicine

基　　金：国家自然科学基金（81460194）;广西自然科学基金（2012GXNSFAA053082）;广西壮族自治区卫生厅自筹经费科研课题（Z2012097）;广西高等学校科研项目（201204LX050）

摘　　要：目的探讨载脂蛋白E（ApoE）对实验性自身免疫脑脊髓炎（EAE）白介素17（IL-17）、白介素6（IL-6）、肿瘤坏死因子-α（TNF-α）表达的影响。 方法选取10只ApoE基因敲除的C57BL/6J小鼠设为ApoE基因敲除组（ApoE-/-组），同时将40只正常C57BL/6J小鼠按随机数字表法分成野生组（WT组）、ApoE拟肽（COG1120）治疗组、生理盐水组和正常对照组，每组10只。ApoE-/-组、WT组、COG112治疗组和生理盐水组均采用背部皮下多点注射完全抗原（MOG35-55的PBS缓冲液与等量的含卡介苗的完全弗氏佐剂混合而成）方法诱导制作EAE模型，正常对照组用生理盐水代替MOG35-55。COG112治疗组在第2-35天每隔2天背部皮下注射COG112 5 mg/（kg·d），生理盐水组用生理盐水代替COG112。在第0-35天对各组小鼠进行神经功能缺损评分；第35天评分后大脑和脊髓取材，应用HE染色观察炎性细胞浸润情况，应用免疫组化染色检测IL-17、IL-6、TNF-α表达水平。结果ApoE-/-组神经功能缺损峰值评分较WT组明显增高，COG112治疗组神经功能缺损峰值评分较生理盐水组明显降低，差异均有统计学意义（P〈0.05）。ApoE-/-组的炎性细胞浸润程度重于WT组，COG112治疗组的炎性细胞浸润程度较生理盐水组轻。ApoE-/-组、WT组、COG112治疗组和生理盐水组小鼠大脑和脊髓IL-17、IL-6、TNF-α平均吸光度值均明显高于正常对照组，差异均有统计学意义（P〈0.05）；ApoE-/-组小鼠大脑和脊髓IL-17、IL-6、TNF-α平均吸光度值均明显高于WT组，COG112治疗组小鼠大脑和脊髓IL-17、IL-6、TNF-α平均吸光度值均明显低于生理盐水组，差异均有统计学意义（P〈0.05）。结论ApoE缺乏会加重EAE炎性细胞浸润及增加IL-17、IL-6、TNF-α表达；ApoE拟肽的干预可减少炎性细胞浸润及IL-17、IL-6、TNF-α表达，从而在EAE的免疫炎症反应中起保护作用。Objective To explore the effect of apolipoprotein E （ApoE） on expressions of interleukin （IL）-IT, IL-6 and tumor necrosis factor （TNF）-α in central nervous system of mice with experimental autoimmune encephalomyelitis （EAE）. Methods Ten C57BL/6J ApoE-deficient （ApoE-/-） mice were selected randomly as ApoE-/- group; 40 normal C57BL/rJ mice were randomly divided into wild-type （WT） group, peptide （COG1120）-treated group, vehicle group and normal control group （n=10）, and mice in the ApoE-/- group, WT group, COGll20-treated group and vehicle group were injected with completeantigen （MOG35-55 in an PBS buffer with complete Freund adjuvant containing Bacillus Calmette-Guerin vaccine） to induce EAE models. Mile in normal control group were not induced EAE models. Mice of COG1120-treated group were injected subcutaneously with COG112 （5 mg/kg body weight in saline） every two days from day 2 to day 35 postimmunization. Mice of vehicle group were injected with physiological saline instead of COG112. Neurological severity scale scores from day 0 to day 35 were recorded. On day 35 after immunization, mice were sacrificed; spinal cord and brain tissues were harvested; the inflammatory infiltration was observed through HE staining; the expressions ofIL-17, IL-6 and TNF-α were quantified by immunohistochemistry. Results The peak symptoms at neurological severity scale and degrees of inflammatory cell infiltration in ApoE-/- group were significantly worse than those in WT group （P〈0.05）. The average optical density of IL-17, IL-6 and TNF-α in the ApoE-/- group, WT group, COG1120-treated group and vehicle group was significantly higher than that in the normal control group （P〈0.05）; that in the ApoE-/- group was significantly higher than that in the WT group （P〈0.05）; that in the COG1120-treated group and was significantly higher than that in the vehicle group （P〈0.05）. Conclusions ApoE deficiency in the EAE mile could enhance inflammatory infiltrates into CNS, a

关 键 词：实验性自身免疫脑脊髓炎 载脂蛋白E 白介素17 白介素6 肿瘤坏死因子-Α 

分 类 号：R744.51[医药卫生—神经病学与精神病学]