摘要
目的:探讨氧化修饰低密度脂蛋白(ox-LDL)对人单核细胞源树突状细胞(DC)功能的影响。方法:采用免疫磁珠法分离人外周血CD14+单核细胞,经含rhGM-CSF(100μg/L)和rhIL-4(20μg/L)的Cellgro培养,使其分化为DC。DC与100 mg/L天然的或氧化修饰的LDL孵育72 h后,采用透射电镜和尼罗红染色观察细胞内脂质沉积,流式细胞术检测DC表型(CD1 a,CD40,CD86,HLA-DR),混合T淋巴细胞反应检测DC对淋巴细胞增殖的影响,FITC-dextran检测DC吞噬功能,ELISA检测细胞培养上清Th1/Th2(IL-12/IL-2)细胞因子的浓度。结果:ox-LDL可诱导DC形成泡沫细胞,而天然的LDL无此作用。经ox-LDL处理的DC吞噬作用明显弱于天然的LDL,而对T细胞增殖作用却明显强于天然的LDL;可明显上调CD80(72.4±9.6vs89.5±10.1,P<0.01),CD86(67.2±8.8vs80.2±11.6,P<0.01),HLA-DR(80.6±9.8vs86.6±10.8,P<0.01)和CD1 a(40.2±10.3vs60.2±9.3,P<0.01)的表达,明显促进DC细胞因子IL-12[(44.3±8.9)ng/Lvs(65.1±10.4)ng/L,P<0.05]的分泌,但却降低IL-2[(43.6±7.8)ng/Lvs(10.0±4.5)ng/L,P<0.01]。结论:DC可通过摄取ox-LDL形成泡沫细胞,而后者与成熟DC的功能相似,说明DC可能是泡沫细胞新的来源,在动脉粥样硬化免疫病理发生中具有重要的作用。
AIM： The purpose of this study is to investigate the mechanisms related to oxidized low - density lipoprotein （ox- LDL） and dendritic cells （DCs） in the process of atherosclerosis. METHODS： Human DCs were prepared from human CD14^＋ peripheral blood monocytes using rhGM - CSF （ 100 μg/L） and rhlL -4 （40 μg/L）. Cells were incubated with 100 mg/L native or oxidized LDL for 72 h. The formation of foam cells was investigated by electron microscopy and oil red O staining. Phenotypic and immune functional assays were used with FACS, FITC -dextran phagocytosis, allogeneic mixed T lymphocytes reaction and secretion of Th1/Th2 （IL- 12/IL- 2） cytokines were also conduced. RESULTS ： DCs treated with ox - LDL, but not native LDL were induced into foam cells after cultured for 72 h. Compared with native LDL, ox - LDL - treated DCs were less potent in FITC - dextran phagocytosis, ox - LDL promoted allogeneic T cells proliferation. Moreover, ox - LDL upregulated CD80 （72. 4 ± 9.6 vs 89.5 ± 10. 1, P 〈0.01 ） , CD86 （67.2 ± 8. 8 vs 80.2±11.6, P〈0. 01）, HLA-DR （80.6±9.8 vs 86.6±10.8, P〈0.01） and CDla （40.2±10.3 vs60.2±9.3, P 〈 0.01 ） expressions, increased IL - 12 secretion [ （44. 3 ± 8.9 ） ng/L vs （65. 1 ± 10. 4 ） ng/L, P 〈 0. 05 ] in DCs. However, the secretion of IL - 2 was decreased [ （43.6 ± 7.8 ） ng/L vs （ 10. 0 ± 4. 5 ） ng/L, P 〈 0. 01 ] significently. CONCLUSION ： DCs were induced into foam cells by ingesting ox - LDL with some functional characteristic of mature DC. DCs seem to be a new source of foam cells and play a key role in immunopathogenesis of atherosclerosis.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2006年第10期1931-1934,共4页
Chinese Journal of Pathophysiology
基金
国家重点基础研究发展规划资助项目(No.G2000056903)
上海市科委资助项目(No.02JC14026)
