摘要
目的观测氧化型低密度脂蛋白（OX—LDL）对外周血单核来源的树突状细胞（DC）成熟及功能的影响，初步探讨其作用机理。方法体外分离培养外周血单核来源的DC，按照处理方式不同分为4组，分别给予磷酸缓冲液（PBS）、低密度脂蛋白（LDL）、ox-LDL、肿瘤坏死因子α（TNF-α）刺激。随后进行流式细胞分析各组DC表面分子的表达情况。使用ELISA方法对培养上清中DC分泌的细胞因子和趋化因子进行测定。使用FITC标记的葡聚糖检测DC吞噬抗原能力变化。使用Westernblot方法对DC胞质内核因子κB（NF—κB）、NF—κB抑制蛋白α（IκBα）的表达进行测定。结果同空白对照组相比，经过ox—LDL处理后的外周血单核来源DC表达CD40（22．3％比45．6％）、CD86（25．9％比82．4％）均明显高，白介素12（31．43pg／ml比126．73pg／ml）、单核细胞趋化蛋白1（59．6ng／ml比116．3ng／ml）分泌多，单核细胞炎性蛋白（MIP1）分泌无明显变化。同时处理后的DC抗原吞噬能力低（46．8％比10．7％），激活自体淋巴细胞增殖能力强（刺激指数4．5比5．7），免疫蛋白印迹实验表明，经ox—LDL处理后DC胞质IκBα的降解多，NF—κB表达高。结论ox—LDL可以促进外周血单核来源的DC成熟，其机制与NF-κB抑制蛋白IκBα降解有关。
Objective To observe the effects of ox-LDL on monocyte-derived dendritic cells. Methods DCs were derived from healthy donors and divided into four groups according to the method of stimulation. The cells of blank group, negative group, experimental group and positive group which were treaded with PBS, LDL, ox-LDL, TNF-α, individually, ox-LDL was added during the late stage of monoeyte differentiation. Flow cytometry was used to analyze the cell surface markers and the endocytes of DCs. 3 H-TdR incorporation was used to measure the proliferation of syngeneie and allogeneie T cells. ELISA assay was used to measure IL-12 ,MCP-land MIP1 in cultured medium. Western blot analysis was used to evaluate the content of IκBα and NF-κB of DCs. Results Addition of ox-LDL during the late stage of monoeytes differentiation can upregulate the cell surface markers including CD40 （22. 3% vs. 45.6% ） and CD86 （25.9% vs. 82.4% ）, increase the secretion of IL-12（31.43 pg/ml vs. 126.73 pg/ml） and MCP-1（59.6 ng/ml vs. 116. 3 ng/ml）, reduce DCs uptake capacity（46. 8% vs. 10. 7% ）, enhance allogeneic Y cells proliferation（SI： 4. 5 vs. 5.7）, promote IκBα degradation and upregulate the expression of NF-κB in DCs. Conclusion ox-LDL can promote the maturation of PBMCs-derived DCs by promoting IκBα degradation.
出处
《中华心血管病杂志》
CAS
CSCD
北大核心
2008年第12期1106-1109,共4页
Chinese Journal of Cardiology
