【摘要】 目的探究载脂蛋白E(Apo E)对髓鞘抗原刺激的小鼠脾脏淋巴细胞基质金属蛋白酶-9(MMP-9)表达的影响。方法以髓鞘少突胶质细胞糖蛋白多肽35-55(MOG35-55)作为抗原诱导免疫Apo E基因缺失型(Apo E-/-)和野生型(WT)型的C57BL/6J小鼠,免疫后第7天取小鼠脾脏,分离并体外培养淋巴细胞,随机分为4组,即Apo E-/-实验组(E-Apo E-/-)、WT实验组(E-WT)、Apo E-/-对照组(C-Apo E-/-)、WT对照组(C-WT)。予以MOG35-55刺激E-Apo E-/-组和E-WT组淋巴细胞,C-Apo E-/-组和C-WT组给予等量生理盐水作为对照处理,24 h后离心并收集上清。采用酶联免疫吸附实验(ELISA)测定各组细胞分泌肿瘤坏死因子-α(TNF-α)、白细胞介素-17(IL-17)、基质金属蛋白酶-9(MMP-9)、基质金属蛋白酶组织抑制剂-1(TIMP-1)的浓度。结果 (1)小鼠被诱导免疫后,Apo E-/-小鼠和WT小鼠脾脏淋巴细胞均能分泌TNF-α、IL-17、MMP-9,Apo E-/-小鼠脾脏淋巴细胞分泌的量较多,差异有统计学意义(P<0.05);(2)体外用MOG35-55刺激脾脏淋巴细胞后,E-Apo E-/-组和E-WT组淋巴细胞TNF-α、IL-17、MMP-9的浓度明显升高,且E-Apo E-/-组较E-WT组更显著,差异有统计学意义(P<0.05);(3)各组小鼠脾脏淋巴细胞分泌TIMP-1的量未见明显差别,差异无统计学意义(P>0.05)。结论 Apo E可以调节脾脏淋巴细胞分泌炎症因子TNF-α、IL-17和基质金属蛋白酶MMP-9,进而可能影响血脑屏障的完整性。更多还原

【Abstract】 Objective To investigate the effect of apolipoprotein E( Apo E) on the expression of matrix metalloproteinase-9( MMP-9) in lymphocytes. Methods Myelinated oligodendrocyte glycoprotein polypeptide 35-55( MOG35-55) antigen induces immunization of Apo E deficient( Apo E-/-) and wild-type( WT) of C57 BL/6 J mice. The spleen of mice was removed on the 7 th day after immunization. Lymphocytes were isolated and cultured in vitro and were randomly divided into four groups: Apo E-/-experimental group( E-Apo E-/-),WT experimental group( E-WT),Apo E-/-control group( C-Apo E-/-) and WT control group( C-WT). The MOG35-55 stimulated the E-Apo E-/-group and the E-WT group lymphocytes,C-Apo E-/-group and C-WT group were treated with equal volume of saline as control. After 24 hours,the supernatant was collected by centrifugation. ELISA was performed to detect the concentrations of tumor necrosis factor-α( TNF-α),interleukin-17( IL-17),matrix metalloproteinase-9( MMP-9),tissueinhibitor of metalloproteinases 1( TIMP-1). Results( 1) After the mice were induced to immunize,the splenic lymphocytes in Apo E-/-mice and WT mice could secrete TNF-α,IL-17 and MMP-9,the amount of secretion of Apo E-/-mice was more significant,and the difference was statistically significant( P < 0. 5).( 2) The concentrations of TNF-α,IL-17 and MMP-9 in splenic lymphocytes of E-Apo E-/-group and E-WT group were significantly increased after two kinds of mouse splenic lymphocytes were stimulated with MOG35 ~ 55 in vitro,and the E-Apo E-/-group was more significant than the E-WT group( P < 0. 05),the difference was statistically significant( P < 0. 05).( 3) There was no significant difference in the concentration of TIMP-1 in each group of splenic lymphocytes,and the difference was not statistically significant( P >0. 05). Conclusion Apo E may affect the integrity of the blood-brain barrier by regulating spleen lymphocyte secretion of inflammatory factors TNF-α、IL-17 and Matrix metalloproteinase MMP-9.更多还原