中文摘要：

目的 观察Toll样受体（TLR）4是否参与氧化型低密度脂蛋白/β2糖蛋白L/抗β2糖蛋白Ⅰ（ox-LDL/β2 GPI/抗β2GPI）抗体复合物诱导小鼠腹腔巨噬细胞泡沫化以及相关因子的表达,以探讨TLR4的作用.方法 分离C3 H/HeN（TLR4正常）和C3 H/HeJ（TLR4缺陷）小鼠腹腔巨噬细胞后,根据实验分组分别加入ox-LDL、ox-LDL/β2 GPI、ox-LDL/抗β2GPI、β2GPI/抗β2GPI、ox-LDL/β2GPI/抗β2 GPI、脂多糖（LPS）处理48 h,油红O染色观察C3H/HeN和C3H/HeJ 2组细胞泡沫化情况；收集刺激后2组细胞总RNA及总蛋白,荧光定量聚合酶链式反应检测2组细胞组织因子（TF） mRNA水平,Western blot检测细胞核因子（NF）-κB p65的活化,酶联免疫吸附法检测细胞因子单核细胞趋化蛋白-1（MCP-1）表达,采用单向方差及双向方差分析组内及组间差异.结果 （1）C3H/HeN小鼠组腹腔巨噬细胞经ox-LDL/β2GPI/抗β2GPI抗体复合物刺激后胞质中脂滴显著增加,与C3 H/HeJ小鼠组比较差异明显.（2） ox-LDL/β2 GPI/抗β2GPI抗体复合物诱导C3 H/HeN小鼠组腹腔巨噬细胞泡沫化过程中,磷酸化-NF-κB p65表达增加,与其空白对照比较差异有统计学意义（P＜0.01）,与ox-LDL/β2GPI/抗β2GPI抗体复合物诱导的C3H/HeJ组比较,差异也有统计学意义（0.82 ±0.22比0.57±0.32,P ＜0.01）.（3）ox-LDL/β2 GPI/抗β2GPI抗体复合物诱导C3H/HeN小鼠组巨噬细胞源性泡沫细胞形成时,TF mRNA及MCP-1表达明显增加,与其空白对照比较差异有统计学意义（P＜0.01）,与ox-LDL/β2 GPI/抗β2GPI抗体复合物诱导的C3 H/HeJ组比较,差异也有统计学意义[TF mRNA：0.041±0.023比0.005±0.003;MCP-1：（6 200.0±6.4）pg/ml比（803.3±5.5）pg/ml,P均＜0.01].结论 TLR4能够促进ox-LDL/β2GPI/抗β2GPI抗体复合物诱导小鼠巨噬细胞源性泡沫细胞的形成,并促进相关分子的表达及活化,提示TLR4在抗磷脂综合征相关分子诱导巨噬细胞泡沫化过程中发挥重要作用.

英文摘要：

Objective To explore the role of toll-like receptor 4 （TLR4） on oxidized low-density/ 132-glycoprotein I/β2-glyeoprotein I （ox-LDL/β2GPI/antiqβ2GPI） antibodies complex induced maerophage foam cell formation. Methods The peritoneal macrophages were separated from TLR4 intact C3H/HeN mice and TLR4 defective C3H/HeJ mice. The cells were treated with ox-LDL, ox-LDL/β2GPI, ox-LDL/ anti-β2 GPI, anti-β2 GPI/β2 GPI, ox-LDIMβ2 GPI/anti-β2 GPI, lipopolysaccharide （LPS） for 48 h and the foam cells were identified by Oil red O staining for intracellular lipids. The total cellular RNA and the protein lysates were collected. The levels of tissue factor （TF） mRNA in two groups were detected by real- time PCR （RT-PCR） , and the expression of phosphorylated nuclear factor-κB （NF-κB） 1365 was detected by Western blotting. Monocyte chemotaetie protein-1 （ MCP-1 ） secretion from peritoneal macrophages was determined by enzyme linked immunosorbent assay （ELISA） kits. Results Compared with C3H/HeJ mice, lipid droplets in the cytoplasm of peritoneal macrophages from C3H/HeN mice were significantly increased and phosphorylation-NF-κB expression was significantly upregulated after stimulating by ox-LDIf β2 GPI/anti-β2 GPI complex（P 〈0. 01 ）. TF mRNA and MCP-1 expression were also upregulated post ox- LDL/β2GPI/anti-β2GPI complex stimulation [TF mRNA： 0. 041 ± 0. 023 vs. 0. 005 ± 0. 003 ; MCP-1 ： （6 200.2±6.4） pg/ml vs. （803.3 ±5.5） pg/ml, P〈0. 01]. Conclusion TLR4 can enhance ox-LDIM β2 GPI/anti-β2 GPI complex induced peritoneal maerophage foam cell formation via upregulating phosphorylation-NF-KB, TF and MCP-1 expression.