不同浓度次级淋巴组织趋化因子对溃疡性结肠炎大鼠淋巴细胞迁移的影响及其意义  

Impact of secondary lymphoid tissue chemokine on lymphocyte migration and the significance thereof in pathogenesis of ulcerative colitis： experiment with rats

作　　者：葛步军[1] 陈锡美[2] 杨长青[2] 吴剑[2] 

机构地区：[1]同济大学附属同济医院外科,上海200065 [2]同济大学附属同济医院消化科,上海200065

出　　处：《中华医学杂志》2008年第44期3112-3115,共4页National Medical Journal of China

基　　金：国家自然科学基金资助项目（30470794）

摘　　要：目的探讨不同浓度次级淋巴组织趋化因子（SLC）对溃疡性结肠炎（UC）大鼠淋巴细胞迁移能力的影响及意义。方法60只大鼠分为UC模型组（模型组）、SLC抗体干预组（干预组）、对照组，各20只，采集各组大鼠门静脉血，分离出淋巴细胞培养，RT—PCR检测淋巴细胞趋化因子受体CCR7的表达，利用Boyden小室法探讨不同浓度SLC对不同组别淋巴细胞的体外趋化作用，酶联免疫吸附试验（ELISA）检测淋巴细胞培养上清液中自细胞介素（IL）-10和干扰素（IFN）-γ的表达。结果3组大鼠淋巴细胞均能表达趋化因子受体CCR7，模型组和干预组CCR7mRNA的表达量（0．792±0．108、0．386±0．115）均明显高于对照组（0．106±0．029，均P〈0．01）；SLC对模型组的淋巴细胞有明显的趋化活性，并呈现剂量效应关系，80ng／ml为SLC的饱和浓度，此浓度下可提高淋巴细胞的迁移能力，模型组与对照组相比，差异有统计学意义（迁移细胞数85．9±16．0vs20．5±1．8，P〈0．01）；干预组迁移细胞数（38．2±6，3）也高于对照组（P〈0．05）；淋巴细胞分泌IL-10的水平明显低于对照组（60±16vs195±39，P=0．036），IFN-γ的分泌明显高于对照组（150±26vs51±13，P=0．042）。结论SLC促进淋巴细胞的迁移，增强SLC诱导淋巴细胞细胞分化，参与UC的发病。Objective To explore the impact of secondary lymphoid tissue chemokine （SLC） on lymphocyte migration and the significance thereof in the pathogenesis of ulcerative colitis （UC）. Methods Sixty SD rats were randomly divided into 3 equal groups： model group undergoing dripping of 40% acetone solution of dinitro-chlorobenzene （DNCB） on the back for 2 weeks and then enema of 6% DNCB acetone solution so as to establish models of UC, and then intravenous injection of normal saline （NS） for 5 days; SLC antibody intervention group undergoing intravenous injection of SLC antibody 15 μg · ml^-1· kg^-1 immediately after the establishing of model; and control group undergoing enema of NS nly and then intravenous injection of NS for 5 days. Six days after the establishing of model venous blood samples were collected from the portal veins of the 3 groups. Lymphocytes were isolated and cultured. RT-PCR was used to detect the mRNA expression of the SLC receptor CCR7. Boyden chamber system was used to examine the migration ability of the lymphocytes exposed to SLC of 20, 40, 60, 80, and 100 ng/ml respectively. ELISA was used to detect the expression of interleukin （ IL）-10 and interferon （IFN） -γ in the supernatants of the lymphocytes of different groups. Results RT-PCR showed that the CCR7 mRNA expression level of the model group was （0. 792 ±0. 108）, significantly higher than that of the intervention group （0. 386 ±0. 115, P = 0. 0429） , and the CCR7 mRNA expression levels of these 2 groups were both significantly higher than that of the control group （0, 106 ± 0. 029, both P 〈 0.01 ）. SLC dose-dependently promoted the migration ability of the lymphocytes, but there existed a saturation phenomenon. Exposed to 80 ng/ml SLC the migration level of the lymphocytes of the model group peaked to （85.9 ± 16. 0）, 3.7 times as high as that of the control group （20. 5 ± 1.8, P 〈 0.01 ）, and the migration level of the lymphocytes of the intervention group was 38.2 ± 6. 3, significa

关 键 词：趋化因子类 结肠炎 溃疡性 淋巴细胞 逆转录聚合酶链反应 

分 类 号：R686[医药卫生—骨科学]