摘要
背景:趋化因子受体及其配体是白细胞聚集的关键因素,可介导淋巴细胞聚集到正常的小肠。目的:研究趋化因子受体CCR2、CCR5及其配体巨噬细胞炎性蛋白(MIP)-1α在小鼠克罗恩病(CD)肠黏膜固有层单个核细胞(LPMC)中的表达,探讨炎症性肠病(IBD)发生、发展的分子机制。方法:健康雄性BALB/c小鼠20只,随机分为对照组和模型组。模型组小鼠以三硝基苯磺酸(TNBS)灌肠造模。5d后处死,提取和培养LPMC,采用流式细胞仪测定小鼠LPMC中趋化因子受体CCR2、CCR5的表达;采用酶联免疫吸附测定(ELISA)检测培养上清液中MIP-1α的分泌;采用逆转录聚合酶链反应(RT-PCR)检测MIP-1α mRNA的表达。结果:小鼠CD模型组LPMC中CCR2、CCR5阳性CD4+细胞比例显著高于对照组(P<0.05);LPMC原代培养后,模型组上清液MIP-1α的分泌显著高于对照组(P<0.05);模型组MIP-1α mRNA的表达显著高于对照组(P<0.05)。结论:趋化因子受体CCR2、CCR5及其配体MIP-1α在小鼠CD的LPMC中表达增高,可能参与了CD的免疫应答和炎症反应。
Background: Chemokine receptors and their ligands play a key role in the recruitment of leucocytes and can mediate the recruitment of lymphocytes to normal small intestine. Aims: To determine the expression of chemokine receptors CCR2, CCR5 and their ligand macrophage inflammatory protein (MIP)-1α in the colonic lamina propria mononuclear cells (LPMC) of mouse with induced colitis and to explore the molecular mechanism of the development of inflammatory bowel disease (IBD). Methods: Twenty healthy male BALB/c mice were divided into two groups, model group and normal control group. Crohn's disease was induced by trinitrobenzene sulfonic acid (TNBS) in the model group. The mice were sacrificed 5 days afterwards and LPMC were isolated and cultured. The expression of CCR2 and CCR5 on LPMC were determined by flow cytometry. The secretion of MIP-1α in the supernatant was determined by enzyme-linked immunosorbent assay (ELISA), and the expression of MIP-1α mRNA was determined by reverse transcriptase polymerase chain reaction (RT-PCR). Results: The ratio of CCR2- and CCR5-positive CD4^+ cells in LPMC was significantly higher in the model group than that in the normal control group (P〈0.05). The secretion of MIP-1α in the supernatant of primary culture of LPMC was significantly higher in the model group than that in the normal control group (P〈0.05). The expression of MIP-1α mRNA was also significantly higher in the model group than that in the normal control group (P〈0.05). Conclusions: Expression of chemokine receptors CCR2 and CCR5 and their ligand MIP-1α increase significantly in LPMC of mouse model with Crohn's disease, and it may be involved in the immune response and inflammatory reaction of Crohn's disease.
出处
《胃肠病学》
2007年第6期349-352,共4页
Chinese Journal of Gastroenterology