本文選題：氧化型低密度脂蛋白 + 樹突狀細胞　； 參考：《中國動脈硬化雜志》2017年03期

【摘要】：目的應用小鼠高脂模型及樹突狀細胞株研究血凝素樣氧化型低密度脂蛋白受體1(LOX-1)高表達樹突狀細胞(DC)在動脈粥樣硬化中的作用。方法氧化型低密度脂蛋白(ox-LDL)誘導原代培養(yǎng)的C57小鼠骨髓來源的樹突狀細胞(BMDC),采用Western blot檢測BMDC LOX-1蛋白水平;采用流式細胞術(shù)檢測高表達LOX-1和低表達LOX-1細胞亞群的比例;MACS磁珠分選LOX-1表達水平不同的兩種細胞亞群,觀察ox-LDL對兩種細胞亞群的影響及釋放炎癥因子的影響;采用C57小鼠高脂模型,在高脂喂養(yǎng)的不同時間(4周、6周、8周)檢測小鼠總膽固醇水平和主動脈中DC數(shù)量;用不同濃度的Dil標記的ox-LDL(Dil-ox-LDL)刺激DC2.4細胞,熒光顯微鏡觀察各組細胞吞噬作用的差異,Western blot檢測LOX-1的表達。結(jié)果 DC可分為高表達和低表達LOX-1兩種細胞亞群,且ox-LDL能增加LOX-1~(high)DC的比例;分選后的陽性細胞被ox-LDL刺激后,TNF-α和IL-1βmRNA的表達明顯上調(diào)(P0.01),且陽性細胞的上升比例高于陰性細胞。在C57小鼠高脂模型中,高脂喂養(yǎng)組總膽固醇水平明顯升高,且隨著總膽固醇水平的升高,小鼠主動脈中DC增多,且LOX-1~(high)DC比例明顯增加。用不同濃度的Dil-ox-LDL刺激DC2.4細胞,隨著Dil-ox-LDL濃度的升高,DC2.4細胞對ox-LDL的吞噬也增加,并且ox-LDL可以誘導DC2.4細胞表面LOX-1表達,20 mg/L和40 mg/L的ox-LDL對LOX-1表達的誘導作用明顯。結(jié)論 ox-LDL能夠上調(diào)DC表面受體LOX-1,增加LOX-1~(high)DC比例,促進炎癥因子表達。
[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- 偽 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.
【作者單位】： 第二軍醫(yī)大學藥學院藥理學教研室;
【基金】：國家自然科學基金資助項目(30972713)
【分類號】：R54

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