胆汁酸代谢异常对Raw264.7细胞功能的影响

Effects of Abnormal Bile Acid Metabolism on the Function of Raw264.7 Cells

目的 探讨异常代谢的胆汁酸单独或联合细菌脂多糖(LPS)对小鼠单核/巨噬细胞系Raw264.7活力、极化及吞噬功能的影响。方法 采用CCK-8法检测不同浓度、时间点鹅去氧胆酸(CDCA)、胆酸(CA)和脱氧胆酸(DCA)单独或联合LPS对Raw264.7细胞活力的影响;CD86和CD163分别标记M1及M2型巨噬细胞,流式细胞仪检测胆汁酸单独或联合LPS处理对Raw264.7极化和吞噬荧光素标记的右旋糖酐(FITC-Dextran)功能的影响。结果 100 ng·mL^(-1) LPS刺激细胞4 h时,可显著增加细胞活力,促进细胞发生M1型极化,并增加吞噬功能(P<0.05)。随着CDCA和DCA刺激的时间延长和浓度上升,细胞活力显著下降(P<0.05),单独使用CDCA刺激巨噬细胞对极化和吞噬功能没有影响,与LPS联合作用介导巨噬细胞发生M1型极化,吞噬功能下降(P<0.05);100μmol·L^(-1) CA刺激细胞后,细胞活力显著升高(P<0.05),单独使用CA刺激细胞对极化和吞噬功能没有影响,与CA组比较,CA与LPS联合作用,介导细胞发生M1型极化,吞噬能力增强(P<0.05)。结论 单独作用情况下,DCA下调细胞吞噬功能,CA、CDCA对巨噬细胞的极化和吞噬功能均无明显影响;但在LPS诱导的体外细胞模型中,CA、CDCA、DCA参与调控巨噬细胞活性、极化以及吞噬功能。

Objective To probe into the effects of abnormally metabolized bile acids alone or in combination with lipopolysaccharide(LPS)on the cell viability,polarization,and phagocytic function of mouse monocyte/macrophage line Raw264.7.Methods CCK-8 method was used to determine the effects of chenodeoxycholic acid(CDCA),cholic acid(CA)and deoxycholic acid(DCA)alone or combined with LPS on Raw264.7 viability at different concentrations and times.M1 and M2 macrophages were labeled with CD86 and CD163,respectively.The effects of bile acids alone or combined with LPS on the polarization and phagocytosis of FITC-dextran of Raw264.7 were detected by flow cytometry.Results Stimulation of cells with 100 ng·mL^(-1) LPS for 4 h could significantly increase cell viability,promote M1 polarization,and increase phagocytosis(P<0.05).With the increase in the time and concentration of CDCA and DCA stimulation,the cell viability decreased notably(P<0.05).The phenotype and phagocytic function of macrophages stimulated by CDCA alone did not change significantly,while combined action with LPS mediated macrophage polarization to the M1 phenotype and decreased phagocytosis(P<0.05).When the cells were stimulated by 100μmol·L^(-1) of CA,the cell viability increased significantly(P<0.05).CA stimulation alone did not affect polarization and phagocytosis,while the stimulation of CA combined with LPS mediated M1 polarization and enhanced phagocytosis(P<0.05).Conclusion Under the condition of individual action,DCA downregulated the phagocytic function of macrophages,while CA and CDCA had no significant effect on macrophages'polarization and phagocytic function.In the LPS induced cell model in vitro,CA,CDCA,and DCA regulate macrophage viability,polarization,and phagocytosis.