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内质网应激通过SCAP/SREBP-1c调控L02肝细胞脂质合成代谢

, PP. 443-448

Keywords: 非酒精性脂肪肝,内质网应激,SCAP,RNA干扰,SREBP-c

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Abstract:

目的在内质网应激状态下,观察SCAP/SREBP-1c对肝细胞脂质合成代谢的影响。方法用毒胡萝卜素(Tg)诱导人L02正常肝细胞株建立内质网应激模型,Westernblot检测GRP78的蛋白表达。甘油三酯(TG)试剂盒和油红O染色检测肝细胞内脂变程度;实时荧光定量PCR检测SREBP-1c下游脂代谢相关基因FAS、ACC1的mRNA表达情况,Westernblot检测nSREBP-1c、FAS、ACC1的蛋白表达情况。通过miRNA瞬时转染沉默SCAP基因的表达,观察上述指标的变化情况。结果实验组GRP78蛋白的相对表达量与对照组比较,均显著增加(P<0.05),Tg在体外成功建立肝细胞内质网应激模型。与空白对照组相比,Tg组肝细胞内甘油三酯含量明显增加(P<0.05),脂滴明显增多;nSREBP-1c的蛋白水平明显升高(P<0.05);下游靶基因FAS和ACC1的基因和蛋白水平均明显上调(P<0.05),与nSREBP-1c的升高趋势一致。转染SCAP-3miRNA载体后,以上指标均明显下调(P<0.05)。结论内质网应激通过SCAP/SREBP-1c调控肝细胞脂质合成代谢,促进脂质沉积,是NAFLD重要的发病机制之一。靶向干扰SCAP,能减轻内质网应激所致的肝细胞脂肪变程度,成为安全有效的NAFLD防治新靶点。

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