人单核巨噬细胞泡沫化抑制剂筛选模型的建立及应用

Establishment and Application of Screening Model for Inhibitor of Foam Cell Formation from Human Monocyte-derived Macrophages

目的 :建立人单核巨噬细胞泡沫化抑制剂筛选模型 ,筛选得到可抑制细胞泡沫化的抑制剂。方法 :U937单核细胞经 10 0nmol·L-1佛波酯 (PMA)诱导 72h分化为巨噬细胞后 ,换无血清培养液于 96孔板中 ,每毫升含巨噬细胞 1× 10 6个 ,每孔再加入 80mg·L-1氧化的低密度脂蛋白 (ox -LDL) ,37℃培养 4 8h ,建立单核巨噬细胞泡沫化模型。利用微生物发酵液 ,或单一的化合物样品与其共孵育 ,油红。染色后观察细胞胞内变化 ,寻找对泡沫细胞形成有抑制作用的样品。利用基因工程技术表达的人清道夫受体A类II型的胞外部分 ,在本模型中可抑制巨噬细胞泡沫化的形成 ,进一步验证了模型的可行性。结果 :从 2 0 0 0个微生物发酵液中筛选到 6株微生物发酵液为阳性 ,从 10个化合物中发现一个有抑制巨噬细胞泡沫化活性的新化合物。结论

Objective: To discover new drugs which may inhibit the formation of foam cell,an inhibitor screening model using human monocyte derived macrophages was established.Method: Human monocyte cell line U937 was stimulated and transformed to macrophages by 100 nmol/L PMA for 72 hours. The monocyte derived macrophages were transferred to 96 well plate with serum free RPMI 1640 medium. They were incubated with 80 mg·L -1 ox LDL at 37 ℃ for 48 hours. Then foam cell model was established. The fermentation broths of microorganisms or some chemical compounds were added into 96 well plate. The lipid droplet in cells was stained by oil red O. All wells were examined by microscope, looking for the samples that inhibited formation of foam cells.Result: Using this screening model, 6 positive strains were found from 2 000 kinds of fermentation broths of microorganisms. 1 new compound was found to inhibit formation of foam cells from 10 compounds tested by this model.Conclusion: This model can be used on high through put screening for inhibitors of foam cell fermation.