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目的观察尿毒症毒素硫酸吲哚酚(IS)是否促进巨噬泡沫细胞脂滴的蓄积,以及对细胞内B类1型清道夫受体(SR-B1)表达和核因子NF-κB活化的影响。方法体外佛波酯(PMA)诱导人源单核细胞系THP-1分化为巨噬细胞,加入氧化低密度脂蛋白(ox-LDL)培养促进巨噬细胞泡沫化,随后以不同浓度的IS干预巨噬泡沫细胞,油红O染色观察细胞内脂滴变化,实时PCR和Western blotting检测SR-B1 m RNA和蛋白表达的水平,ELISA法检测NF-κB活化情况;经NF-κB抑制剂四氢化吡咯二硫代氨基甲酸酯(PDTC)预处理后IS再干预,实时PCR检测巨噬泡沫细胞内SR-B1 m RNA表达的变化。结果与对照组(IS 0μmol/L)比较,IS干预后巨噬泡沫细胞内脂滴明显增加,随着IS浓度的升高,细胞内脂滴逐渐增加;IS干预后巨噬泡沫细胞的NF-κB活化程度增高,差异有统计学意义(P<0.05)。与对照组比较,细胞在不同浓度的IS(2.5、25、250μmol/L)处理下,SR-B1 m RNA和蛋白的表达下降,随着IS浓度的增加,SR-B1和蛋白逐渐减少,呈现浓度依赖性,差异有统计学意义(P<0.05)。PDTC预处理后SR-B1m RNA表达增加,差异有统计学意义(P<0.05)。结论 IS抑制SR-B1 m RNA和蛋白的表达,增加THP-1源性巨噬泡沫细胞胆固醇的蓄积,促进巨噬细胞泡沫化,从而促进动脉粥样硬化。NF-κB活化可能是IS抑制SR-B1 m RNA表达的可能机制之一。
Objective To investigate whether uremic toxins indoxyl sulfate (IS) promotes the accumulation of lipid droplets in macrophage foam cells and its effect on the expression of type 1 scavenger receptor (SR-B1) and the activation of nuclear factor-kappa B . Methods Phosphatidylcholine (PMA) induced human monocyte cell line THP-1 to differentiate into macrophages. Oxidized LDL (ox-LDL) culture was used to promote the foaming of macrophages, and then with different concentrations of IS intervention The macrophage foam cells and oil red O staining were used to observe the changes of intracellular lipid droplets. The mRNA and protein expression of SR-B1 were detected by real-time PCR and Western blotting. The activation of NF-κB was detected by ELISA. After pretreatment with pyrrolidine dithiocarbamate (PDTC), the expression of SR-B1 m RNA in macrophage foam cells was detected by real-time PCR. Results Compared with the control group (IS 0μmol / L), lipid droplets in macrophage foam cells increased significantly after IS treatment, and lipid droplets increased gradually with the increase of IS concentration. After IS intervention, macrophage foam cells expressed NF- κB activation increased, the difference was statistically significant (P <0.05). Compared with the control group, the expression of SR-B1mRNA and protein decreased with different concentrations of IS (2.5, 25 and 250μmol / L), and the expression of SR-B1 and protein decreased with the increase of IS concentration Concentration-dependent, the difference was statistically significant (P <0.05). The expression of SR-B1m RNA increased after PDTC pretreatment, the difference was statistically significant (P <0.05). Conclusion IS inhibits the expression of SR-B1 m RNA and protein, increases the accumulation of cholesterol in THP-1-derived macrophage foam cells, promotes the foaming of macrophages and promotes atherosclerosis. NF-κB activation may be one of the possible mechanisms by which IS inhibits the expression of SR-B1mRNA.