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目的探讨二甲双胍(Met)对脂多糖(LPS)诱导的THP-1来源泡沫细胞的形成、脂滴形态以及脂滴表面蛋白分子表达的影响。方法采用100 ng/m L佛波酯(PMA)处理THP-1细胞48 h后,诱导其分化成为巨噬细胞;再利用50μg/m L氧化型低密度脂蛋白(ox-LDL)和1μg/m L LPS促进THP-1细胞来源的巨噬细胞形成泡沫细胞,在此过程中用(0、100、200)μmol/L Met进行处理,采用油红O染色分析Met对泡沫细胞形成的影响;采用BODIPY493/503染色,荧光显微镜观察泡沫细胞内脂滴的形态和数量;抽提细胞内脂类,通过定量试剂盒测定细胞甘油三酯(TG)的含量;采用Western blot分析脂滴表面蛋白中脂肪分化相关蛋白(ADRP)、47 k Da的尾连蛋白(TIP47)的表达水平。结果与ox-LDL和LPS组相比,采用100μmol/L、200μmol/L Met处理后能够降低泡沫细胞内的脂滴大小和数量,定量分析显示细胞内TG含量明显降低,并存在剂量依赖关系,其中200μmol/L Met能够降低泡沫细胞中约25%的TG储积。Western blot结果显示,Met能够降低泡沫细胞中ADRP的表达水平,但对TIP47的表达水平无影响。结论 Met能够抑制LPS诱导的THP-1细胞源性泡沫细胞形成,并抑制脂质蓄积,同时下调细胞内ADRP的表达水平。
Objective To investigate the effects of metformin on lipopolysaccharide (LPS) -induced THP-1-derived foam cells formation, lipid droplet morphology and lipid droplet surface protein expression. Methods THP-1 cells were treated with 100 ng / m L phorbol ester (PMA) for 48 h and then induced to differentiate into macrophages. Oxidized low density lipoprotein (ox-LDL) and 1 μg / m L LPS promoted THP-1 cell-derived macrophages to form foam cells. During this process, cells were treated with (0, 100, 200) μmol / L Met Met and the effect of Met on foam cell formation was analyzed by Oil Red O staining. BODIPY493 / 503 staining was used to observe the morphology and quantity of lipid droplets in the foam cells by fluorescence microscopy. The lipids in the cells were extracted and the contents of triglycerides (TG) in the cells were determined by quantitative kit. Adipogenic differentiation related protein (ADRP), 47 kDa connective tissue protein (TIP47). Results Compared with ox-LDL and LPS groups, the amount and amount of lipid droplets in foam cells could be decreased by 100 μmol / L and 200 μmol / L Met Met Met, and quantitative analysis showed that the intracellular TG content was significantly reduced and there was a dose-dependent manner. Among them, 200 μmol / L Met was able to reduce about 25% TG accumulation in foam cells. Western blot results showed that Met can reduce the expression of ADRP in foam cells, but had no effect on the expression of TIP47. Conclusion Met can inhibit LPS-induced THP-1 cell-derived foam cell formation, inhibit lipid accumulation, and down-regulate the expression of ADRP in cells.