ox-LDL促进树突状细胞LOX-1表达及炎症因子分泌

发布时间：2018-05-21 09:59

本文选题：氧化型低密度脂蛋白 + 树突状细胞　；参考：《中国动脉硬化杂志》2017年03期

【摘要】：目的应用小鼠高脂模型及树突状细胞株研究血凝素样氧化型低密度脂蛋白受体1(LOX-1)高表达树突状细胞(DC)在动脉粥样硬化中的作用。方法氧化型低密度脂蛋白(ox-LDL)诱导原代培养的C57小鼠骨髓来源的树突状细胞(BMDC),采用Western blot检测BMDC LOX-1蛋白水平;采用流式细胞术检测高表达LOX-1和低表达LOX-1细胞亚群的比例;MACS磁珠分选LOX-1表达水平不同的两种细胞亚群,观察ox-LDL对两种细胞亚群的影响及释放炎症因子的影响;采用C57小鼠高脂模型,在高脂喂养的不同时间(4周、6周、8周)检测小鼠总胆固醇水平和主动脉中DC数量;用不同浓度的Dil标记的ox-LDL(Dil-ox-LDL)刺激DC2.4细胞,荧光显微镜观察各组细胞吞噬作用的差异,Western blot检测LOX-1的表达。结果 DC可分为高表达和低表达LOX-1两种细胞亚群,且ox-LDL能增加LOX-1~(high)DC的比例;分选后的阳性细胞被ox-LDL刺激后,TNF-α和IL-1βmRNA的表达明显上调(P0.01),且阳性细胞的上升比例高于阴性细胞。在C57小鼠高脂模型中,高脂喂养组总胆固醇水平明显升高,且随着总胆固醇水平的升高,小鼠主动脉中DC增多,且LOX-1~(high)DC比例明显增加。用不同浓度的Dil-ox-LDL刺激DC2.4细胞,随着Dil-ox-LDL浓度的升高,DC2.4细胞对ox-LDL的吞噬也增加,并且ox-LDL可以诱导DC2.4细胞表面LOX-1表达,20 mg/L和40 mg/L的ox-LDL对LOX-1表达的诱导作用明显。结论 ox-LDL能够上调DC表面受体LOX-1,增加LOX-1~(high)DC比例,促进炎症因子表达。
[Abstract]:Objective to study the role of hyperlipidemic mouse model and dendritic cell line (DC) in atherosclerosis by using hemagglutinin like oxidized low density lipoprotein receptor (LOX-1). Methods oxidized low density lipoprotein (ox-LDL) induced BMDC from bone marrow-derived dendritic cells of primary cultured C57 mice, and Western blot was used to detect the level of BMDC LOX-1 protein. Flow cytometry was used to detect the proportion of high expression LOX-1 and low expression LOX-1 cell subsets. Macs magnetic beads were used to separate two kinds of cell subsets with different LOX-1 expression levels. The effect of ox-LDL on the two cell subsets and the release of inflammatory factors were observed. The levels of total cholesterol and the number of DC in aorta of C57 mice were measured at 4 weeks, 6 weeks and 8 weeks after high fat feeding, and the DC2.4 cells were stimulated by ox-LDLL-Dil-ox-LDL labeled with different concentrations of Dil. The difference of phagocytosis in each group was observed by fluorescence microscope. The expression of LOX-1 was detected by Western blot. Results DC could be divided into two subgroups: high expression and low expression LOX-1, and ox-LDL could increase the proportion of LOX-1~(high)DC, and the expression of TNF- 伪 and IL-1 尾 mRNA was up-regulated by ox-LDL, and the percentage of positive cells was higher than that of negative cells. In the model of C57 mice, the total cholesterol level in high fat feeding group increased obviously, and with the increase of total cholesterol level, the DC in the aorta and the proportion of LOX-1~(high)DC in the aorta of the mice increased obviously. When DC2.4 cells were stimulated with different concentrations of Dil-ox-LDL, the phagocytosis of ox-LDL by DC2.4 cells increased with the increase of Dil-ox-LDL concentration, and ox-LDL could induce LOX-1 expression of LOX-1 20 mg/L and 40 mg/L ox-LDL on DC2.4 cells. Conclusion ox-LDL can up-regulate LOX-1, increase the proportion of LOX-1~(high)DC and promote the expression of inflammatory factors.
【作者单位】：第二军医大学药学院药理学教研室;
【基金】：国家自然科学基金资助项目(30972713)
【分类号】：R54

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