【摘要】 目的明确拼接型和非拼接型X盒子结合蛋白1对肾小管上皮细胞脂质代谢的影响。方法体外培养人肾小管上皮细胞,采用脂质体2000转染拼接型和非拼接型XBP-1质粒,培养48h后,免疫细胞化学和Western blot检测XBP-1表达,反转录PCR检测脂肪酸合成酶和乙酰辅酶A羧化酶mRNA表达,免疫细胞化学检测脂肪分化相关蛋白表达,油红O检测细胞内脂滴。结果 HKC细胞转染XBP-1拼接型和非拼接型质粒48h后,免疫细胞化学和Western blot证实XBP-1蛋白均明显升高。而拼接型XBP-1质粒转染上调了FASN和ACC mRNA表达,细胞内ADRP表达升高,细胞内脂滴增多。非拼接型XBP-1质粒对FASN、ACC mRNA,ADRP和细胞内脂滴未产生影响。结论拼接型XBP-1上调可导致肾小管上皮细胞脂质代谢关键酶升高和细胞内脂质沉积,可能是多种肾脏脂质沉积疾病的调控因子之一。更多还原

【Abstract】 Objective To determine the effect of unspliced and spliced XBP-1proteins on lipid metabolism of renal tubular cells.Methods In-vitro cultured HKC cells were respectively transfected with spliced or unspliced XBP-1plasmid by Lipofectamine 2000.After 48 h,XBP-1protein expression was detected by immunocytochemistry and Western blot.Reverse transcript PCR was used to detect FASN and ACC mRNA.ADRP was detected with immunocytochemistry,and intracellular lipid droplets were detected with Oil Red O staining.Results At 48 hafter transfection,both spliced and unspliced XBP-1plasmids increased XBP-1protein expression.Furthermore,spliced XBP-1plasmid caused enhanced FASN and ACC mRNA,high expression of ADRP and intracellular lipid accumulation.Contrarily,unspliced XBP-1plasmid had no effect on FASN,ACC,ADRP and lipid metabolism.Conclusion Spliced XBP-1can increase the key enzymes involved in the lipid metabolism of HKC cells,which may be one of the regulatory mechanisms mediating various renal diseases related with lipid metabolism.更多还原