摘要
目的:体外比较白细胞介素10(IL-10)信号转导机制对C57BL/6和MRL/lpr-小鼠腹腔巨噬细胞功能及活性的影响。方法:分别分离C57BL/6和MRL/lpr-小鼠腹腔巨噬细胞进行培养;用不同浓度IL-10(0.01、0.1、1和10ng/ml)进行刺激,用MTT法测定比较两种品系小鼠腹腔巨噬细胞的增殖反应能力;Real time PCR分别测定其产生前炎性细胞因子IL-1α、M-CSF、TNF-α的量;用100ng/mlIL-10分别刺激两种品系小鼠腹腔巨噬细胞10分钟,免疫印迹法比较两种细胞中JAK1、TYK2、STAT3及磷酸化水平。结果:C57BL/6小鼠巨噬细胞数及其产生的前炎性细胞因子量随IL-10刺激浓度的增加而递减;MRL/lpr-小鼠巨噬细胞数及其前炎性细胞因子IL-1α、M-CSF的量随IL-10刺激浓度的增加而递增,但巨噬细胞产生TNF-α的量随IL-10刺激浓度的增加而递减;MRL/lpr-小鼠细胞内信号转导分子的磷酸化水平低于C57BL/6。结论:MRL/lpr-小鼠的细胞免疫和炎症反应不能被IL-10抑制,可能是由于其信号转导过程中的缺陷所致。
Objective：To compare the effects of recombirrant murine interleukin 10（IL-10）on peritoneal exudated macrophage of C57BL/6 and of MRL/lpr^- mice in vitro. Methods： Peritoneal exudated macrophages were isolated from C57BL/6 and MRL/lpr^- mice, and cultured in .presence of various concentrations （0.01,0.1,1 and 10 ng/ml） of IL- 10 for 72 hours, respectively. Proliferation of the peritoneal cells of both the strains of mice was compared with MTT assay and the expression of proinflammatory cytokines IL-1α, M-CSF and TNF-α was measured with Real time PCR assay. After the cells were stimulated, with 100 ng/ml of IL-10 for 10 min, the amotmt of JAK1, TYK2 and STAT3 and their phosphorylation were detected with Western blot, and compared between C57BL/6 and MRL/Ipr^- mice. Results： With augmented concentration of interleukin 10, the proliferation and secretion of the proinflammatory cytokines of macrophages from C57BL/6 mice were reduced dose-dependently, whereas up-regulated in the ceils from MRL mice except expression of TNF-α, which appeared decreasing. Phospho-rylation degree of the signal transduction molecules of the cells from MRL was lower than that from C57BL/6 mice. Conclusion：The reason that interleukin 10 can not inhibit cell-mediated immtmity and inflammatory reaction in MRL mice may be deficiency of the signal transduetion.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2008年第11期976-979,共4页
Chinese Journal of Immunology
基金
国家自然科学基金(No.30571701、30600541)资助
