【摘要】 目的:观察炎症是否干扰脂肪酸负荷的肾细胞[人系膜细胞(HMCs)和肾小管上皮细胞HK-2]脂肪酸转运蛋白(FAT/CD36)的表达。方法:分别给予不同浓度软脂酸(0 mmol/L、0.02 mmol/L、0.04 mmol/L、0.08 mmol/L、0.16 mmol/L、0.32 mmol/L)处理HMCs和HK-2细胞24 h。采用实时定量PCR和Western blotting方法检测细胞FAT/CD36的mRNA及蛋白的表达。进一步选取0.04 mmol/L软脂酸联合炎症因子(25μg/L TNF-α或20μg/L IL-6)处理肾细胞24 h后,观察炎症因子对肾细胞FAT/CD36 mRNA及蛋白表达的影响;油红O染色及酶比色法检测细胞甘油三酯(TG)水平;ELISA检测细胞游离脂肪酸(FFA)含量。结果:软脂酸呈浓度依赖性上调HMCs和HK-2细胞FAT/CD36 mRNA和蛋白表达。炎症因子明显刺激脂肪酸负荷的肾细胞FAT/CD36mRNA和蛋白表达进一步增加。油红O染色及胞内TG和FFA含量测定显示炎症因子促进肾细胞脂质积聚。结论:炎症上调脂肪酸负荷的肾细胞FAT/CD36表达,加重胞内脂质积聚。更多还原

【Abstract】 AIM: To investigate the effects of inflammatory cytokines on the expression of fatty acid transporter(FAT/CD36) in renal cells loaded by fatty acids.METHODS: Human mesangial cells(HMCs) and renal tubular epithelial HK-2 cells were treated with palmitate at concentrations of 0 mmol/L,0.02 mmol/L,0.04 mmol/L,0.08 mmol/L,0.16 mmol/L and 0.32 mmol/L for 24 h.The expression of FAT/CD36 at mRNA and protein levels was detected by real-time PCR and Western blotting,respectively.The renal cells were treated with palmitate at concentration of 0.04 mmol/L combined with TNF-α(25 μg/L) or IL-6(20 μg/L) for 24 h.The effect of inflammatory cytokines on the mRNA and protein levels of FAT/CD36 in the renal cells was also investigated.Oil red O staining was used to determine the intracellular lipid droplet formation.The intracellular triglyceride(TG) and free fatty acid(FFA) were measured by enzymic assay and ELISA,respectively.RESULTS: Palmitate loading dose-dependently increased the expression of FAT/CD36 at mRNA and protein levels in both HMCs and HK-2 cells.The inflammatory cytokines further increased the expression of FAT/CD36 at mRNA and protein levels in both cells loaded by palmitate.Oil red O staining,TG detection and FFA assay showed that the inflammatory cytokines increased intracellular lipid levels in both HMCs and HK-2 cells.CONCLUSION: Inflammatory cytokines up-regulate the expression of FAT/CD36 in renal cells loaded by fatty acids and exacerbate the intracellular lipid accumulation.更多还原