本文選題：氧化型低密度脂蛋白 + 樹(shù)突狀細(xì)胞��； 參考：《中國(guó)動(dòng)脈硬化雜志》2017年03期

【摘要】：目的應(yīng)用小鼠高脂模型及樹(shù)突狀細(xì)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(dá)樹(shù)突狀細(xì)胞(DC)在動(dòng)脈粥樣硬化中的作用。方法氧化型低密度脂蛋白(ox-LDL)誘導(dǎo)原代培養(yǎng)的C57小鼠骨髓來(lái)源的樹(shù)突狀細(xì)胞(BMDC),采用Western blot檢測(cè)BMDC LOX-1蛋白水平;采用流式細(xì)胞術(shù)檢測(cè)高表達(dá)LOX-1和低表達(dá)LOX-1細(xì)胞亞群的比例;MACS磁珠分選LOX-1表達(dá)水平不同的兩種細(xì)胞亞群,觀察ox-LDL對(duì)兩種細(xì)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(shí)間(4周、6周、8周)檢測(cè)小鼠總膽固醇水平和主動(dòng)脈中DC數(shù)量;用不同濃度的Dil標(biāo)記的ox-LDL(Dil-ox-LDL)刺激DC2.4細(xì)胞,熒光顯微鏡觀察各組細(xì)胞吞噬作用的差異,Western blot檢測(cè)LOX-1的表達(dá)。結(jié)果 DC可分為高表達(dá)和低表達(dá)LOX-1兩種細(xì)胞亞群,且ox-LDL能增加LOX-1~(high)DC的比例;分選后的陽(yáng)性細(xì)胞被ox-LDL刺激后,TNF-α和IL-1βmRNA的表達(dá)明顯上調(diào)(P0.01),且陽(yáng)性細(xì)胞的上升比例高于陰性細(xì)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(dòng)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(xì)胞,隨著Dil-ox-LDL濃度的升高,DC2.4細(xì)胞對(duì)ox-LDL的吞噬也增加,并且ox-LDL可以誘導(dǎo)DC2.4細(xì)胞表面LOX-1表達(dá),20 mg/L和40 mg/L的ox-LDL對(duì)LOX-1表達(dá)的誘導(dǎo)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(jìn)炎癥因子表達(dá)。
[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(xué)藥學(xué)院藥理學(xué)教研室;
【基金】：國(guó)家自然科學(xué)基金資助項(xiàng)目(30972713)
【分類(lèi)號(hào)】：R54

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