髓样分化因子88抑制剂ST2825对RAW264.7细胞生物学活性的影响  

Effects of myeloid differentiation factor 88 inhibitor ST2825 on biological activities of RAW264.7

作　　者：李超[1] 薛承彪[1] 杨涛[1] 李尧[1] 黄霞[1] 张利民[1] 李明强[1] 周平[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院器官移植研究所 卫生部/教育部器官移植重点实验室,武汉430030

出　　处：《中华移植杂志（电子版）》2012年第4期19-22,共4页Chinese Journal of Transplantation(Electronic Edition)

基　　金：国家自然科学基金(81072442)

摘　　要：目的探讨Toll样受体信号转导分子髓样分化因子88(MyD88)抑制剂ST2825对巨噬细胞系RAW264.7细胞生物学活性的影响,为新型免疫抑制剂的研发提供新的思路。方法培养RAW264.7细胞并分为4组:空白对照组细胞不给予任何处理和刺激;脂多糖对照组细胞给予脂多糖(500ng/mL)刺激;ST2825低浓度组和高浓度组细胞分别给予浓度为10μmol/L和20μmol/L的ST2825预处理1h后再给予脂多糖刺激。36h后收集细胞,流式细胞术检测细胞表面共刺激分子CD80和CD86的表达;提取细胞RNA,实时荧光定量PCR检测IL-1、IL-6、TNF-αmRNA水平。异硫氰酸荧光素标记的葡聚糖(FITC-Dextran)法测定细胞吞噬功能。结果脂多糖对照组RAW264.7细胞共刺激分子CD80和CD86表达率分别为(54.2±1.6)%和(56.8±2.9)%,与空白对照组相比均明显升高(均P<0.05);ST2825低浓度组和高浓度组CD80表达率分别为(37.4±1.7)%和(26.4±1.2)%,CD86表达率分别为(43.4±2.0)%和(31.5±1.1)%,与脂多糖对照组相比均降低(均P<0.05)。经脂多糖刺激后,脂多糖对照组RAW264.7细胞IL-1、IL-6和TNF-αmRNA表达水平升高,ST2825可降低上述促炎性细胞因子的mRNA表达水平。ST2825低浓度组和高浓度组细胞吞噬FITC-Dextran的能力与正常RAW264.7细胞无差异。结论 MyD88抑制剂ST2825可抑制脂多糖对天然免疫细胞的免疫活化效应,而不影响细胞的吞噬功能,为ST2825作为免疫抑制剂用于临床奠定了实验基础。Objective To investigate the effect of Toll-like receptor signaling transduction adapter myeloid differentiation factor 88 （MyD88） inhibitor ST2825 on biological activities of RAW264. 7 in costimulatory molecules expression, secretion of cytokines and phagocytic function, and to provide new thinking in the development of new type of immunosuppressive agents. Methods The RAW264.7 cultured in vitro were divided in 4 groups. The normal control group ： the RAW264.7 cell line was not given any stimulation. Lipopolysaccharide（LPS） control group： the RAW264.7 cell line was stimulated with LPS（500 ng/mL）. ST2825 groups： the RAW264.7 cell line was treated with ST2825 1 h before LPS stimulation and the concentration gradient consisted of low concentration （10 μmol/L） and high concentration（ 20 μmol/L）. In 36 h after LPS stimulation, cells were harvested, and then co-stimulatory molecules CD80 and CD86 were analyzed by flow cytometry; IL-1, IL-6, and TNF-α mRNA were detected with real-time PCR; and phagocytic function was analyzed by flow cytometry. Results ST2825 inhibited the upregulation of CD80 and CD86 in a concentration-dependent manner IED80： （37.4±1.7）% vs （26.4±1.2）% and CD86：（43.4 ±2.0）% vs （31.5±1.1）%], and the expression of IL-1, IL-6 and TNF-α mRNA upon the stimulation of LPS in vitro. No significant difference in phagocytic function was noted before and after the administration of ST2825. Conclusion MyD88 inhibitor ST2825 can inhibit the responsive effects of innate immune cells upon LPS stimulation and this study may lay a foundation for clinical application of ST2825 as an immunosuppressive agent.

关 键 词：髓样分化因子88 ST2825 RAW264 7 免疫抑制剂 

分 类 号：R392[医药卫生—免疫学]