一种血清诱导巨噬细胞转化为泡沫细胞新方法的建立

Establishment of a Novel Method to Induce Formation of the Foam Cells from Macrophages with Fetal Bovine Serum

摘要: 目的：利用血清替代氧化型低密度脂蛋白(oxidized low density lipoprotein, ox-LDL)诱导巨噬细胞，建立一种新的诱导泡沫细胞体外培养方法。方法：小鼠腹腔注射RPMI-1640培养液提取巨噬细胞，RPMI-1640无血清培养4 h贴壁，分别加含不同浓度胎牛血清(体积分数5%、10%和20%)的RPMI-1640培养液、无血清氧化型低密度脂蛋白(2 × 10−2 g/L)RPMI-1640培养液培养4 h、8 h和12 h，采用油红O染色，观察巨噬细胞吞噬脂滴转化为泡沫细胞情况。采用酶化学法检测细胞总胆固醇酯和游离胆固醇含量。结果：不同浓度胎牛血清(体积分数5%，10%和20%)作用不同时间(4 h，8 h和12 h)均对巨噬细胞有诱导作用，且随着浓度的增加、时间的延长，巨噬细胞内脂滴增多变大，部分融合成大脂滴。但当血清浓度增加到20%时，作用时间多于8 h，巨噬细胞有部分破裂、脂滴丢失、细胞减少的现象。在浓度为10%的血清作用7 h诱导的泡沫细胞，细胞内的胆固醇酯含量明显高于空白对照的。结论：浓度为10%的血清作用7 h可诱导巨噬细胞转化为泡沫细胞。

Abstract: Objective: To establish a method for forming foam cells from macrophages by serum instead of oxidized low density lipoprotein (ox-LDL) in vitro. Methods: The macrophages were isolated from the mice by intraperitoneal injection RPMI-1640 culture medium. The macrophages were cultured 4 h in serum-free RPMI-1640 medium for adherence. Then, the adherent macrophages were added RPMI-1640 culture medium with different concentrations of fetal bovine serum (volume fraction of 5%, 10% and 20%) or oxidized low density lipoprotein (2 × 10−2 g/L). The cells were stained with oil red O following the above conducts for observing the intracellular lipid droplets. The contents of intracellular total cholesterol and cholesteryl esters were measured by enzymology. Results: Both different concentrations of fetal bovine serum (volume fraction of 5%, 10% and 20%) and different action time (4 h, 8 h and 12 h) can induce the macrophages into foam cells, and the intracellular lipid droplets were bigger and even the cells merged together a bigger lipid droplets. The macrophages membrane was broken and the lipid droplet and cells were lost when the serum was increased to 20% and the action time was more than 8 h. Macrophages cultured by 10% serum and acted 7 h, cellular contents of cholesteroy and cholesteryl ester increased markedly compared with the normal macrophages. Conclusion: A foam cell model has been established by incubating macrophages with 10% fetal-bovine serum amount of RPMI-1640 medium for 7 hours in vitro culture.