中文摘要：

目的探讨Toull样受体4（Toll-1ikereceptor4，TLR4）促进平滑肌源性泡沫细胞的形成是否与三磷酸腺苷-结合盒转运子A1（ATP-binding cassette transporter，ABcA1）表达异常相关。方法组织贴块法培养原代血管平滑肌细胞（vascular smooth muscle cells，VSMCs）；细胞分为野生型（Wildtype）空白对照组、Wildtype氧化低密度脂蛋白（oxLDL）组、TLR4敲除（TLR4-/-）空白对照组和TLR4-/-oxLDL组；油红0染色观察VsMcs内脂质聚集，并用异丙醇萃取油红0测定光密度值；RT-qPCR、western blot检测VSMC8中TLR4、ABcA1、IL-1β及TNF-αmRNA和蛋白表达水平；细胞免疫荧光染色鉴定原代VSMCs及观察核转录因子-kappa（NF-KB）核转位。结果①wild type，oxLDL组中见细胞内大量脂滴形成，油红O光密度值为wild type空白对照组的2．3倍（P〈0．01）；TLR4-/-空白对照组中只见少许脂质沉积于细胞内，油红O光密度值与，TLR4-/-空白对照组相比无明显变化（P〉0．05）。②wild type oxLDL组ABcA1 mRNA及蛋白表达水平较Wild type空白对照组分别下降61％（P〈0．05），44％（P〈0．01）；TLR4-/-oxLDL组ABcA1 mRNA及蛋白表达水平分别为TLR4-/-。空白对照组的2．3倍（P〈0．05），1．7倍（P〈0．05）。③Wild type oxLDL组VsMcs中NF-KB核转位明显增多，IL-1β和TNF-αmRNA表达较wild type空白对照组分别升高3．9倍（P〈0．05），1．7倍（P〈0．05）；TLR4-/- 0xLDL组VsMC8中无明显NF-KB核转位，仅IL-1β mRNA表达较TLR4-/-空白对照组升高1．2倍（P〉0．05）。结论TLR4及其调节的炎性反应参与平滑肌源性泡沫细胞形成过程中ABCA1表达异常。

英文摘要：

Objective To determine whether Toll-like receptor 4 （TLR4） promoting VSMCs-derived foam cells formation is related with the abnormal expression of ATP-binding cassette transporter 1 （ ABCA1 ）. Methods Vascular smooth muscle cells （VSblCs） were isolated from male C57BL/6J （TLR4-/-） and C57BL/lOScNj （ TLR4 -/- ） mice respectively, and primary cultured and identified. The obtained VSblCs were divided intowild type blank control cells, oxLDL-treated wild type cells （50 μg/mL oxLDL）, TLR4-/- blank control cells and oxLDL-treated TLR4 -/- cells. Neutral lipid inclusions in the VSblCs induced by oxLDL were assessed by Oil Red O staining. Lipid accumulation was quantitated by isopropanol extraction of Oil Red O from stained cells and optical density determinations at 518 nm. The mRNA and/or protein levels of TLR4, ABCA1, IL-1 β and TNF-α were determined by Western blotting and RT-qPCR. The NF-KB translocation was identified by immunofluorescence assay. Results Oil red O staining indicated that a large amount of lipid was accumu-- lated in oxLDL-treated wild type VSMCs, with the OD value increased by 2.3 folds compared with that of blank control （ P 〈 0.01 ）. Only few lipids were found in oxLDL-treated TLR4 -/- VSMCs, with no significant difference with the untreated cells （ P 〉 0.05 ）. The ABCA1 expression at mRNA and protein levels wasreduced by 61% （P 〈 0.05 ） and 44% （ P 〈 0.01 ）, respectively, in oxLDL-treated wild type VSMCs than in untreated cells. However, in the TLR4-/- VSMCs stimulated by oxLDL, the ABCA1 mRNA and protein expression were 2.3-fold （ P 〈 0.05 ） and 1.7-fold higher （ P 〈 0.05 ） than the TLR4 -/- blank control, respec- tively. The translocation of NF-KB was activated by TLR4 in oxLDL-treated wild type VSMCs, and the IL-1β and TNF-α mRNA expression were 3.9 folds （P 〈 0.05 ） and 1.7 folds （ P 〈 0.05 ） respectively of wild type blank controls. However, in oxLDL-treated TLR4 knockout VSMCs, NF-KB translocation and the expressi