胰岛素、胰高血糖素对体外培养犊牛肝细胞脂合成作用的影响

发布时间：2018-04-18 02:24

本文选题：胰岛素 + 胰高血糖素　；参考：《中国兽医学报》2017年02期

【摘要】：为确定胰岛素(insulin,INS)和胰高血糖素(glucagon,GLN)对奶牛肝细胞脂合成作用的影响,本试验体外培养犊牛原代肝细胞,分别用不同浓度的INS和GLN处理肝细胞:对照组(0nmol/L)、浓度梯度组(1,10,100,1 000nmol/L)。培养1h后分别用免疫印迹(Western blot,WB)方法、荧光定量PCR(qRT-PCR)方法以及细胞免疫荧光(immunofluorescence,IF)方法检测INS和GLN处理体外培养肝细胞对关键转录因子固醇调节元件结合蛋白-1c(SREBP-1c)mRNA表达水平,SREBP-1c核质分布以及下游靶基因脂代谢关键酶乙酰辅酶A羧化酶(ACC)和脂肪酸合成酶(FAS)mRNA表达水平的影响。结果显示:INS处理使SREBP-1c的表达水平和转录活性显著升高,入核量显著增加,其下游的脂合成关键酶ACC和FAS的基因表达水平也显著升高。而与之相反,GLN处理使SREBP-1c的表达水平和转录活性显著降低,入核量显著减少,其下游的脂合成关键酶ACC和FAS的基因表达水平也显著降低。以上结果说明,INS可通过增强SREBP-1c的活性从而促进肝细胞脂合成,增加肝脂沉积;而GLN则通过抑制SREBP-1c的活性从而降低肝细胞脂合成。
[Abstract]:In order to determine the effect of insulin insulinine (ins) and glucagonine glucagonine (GLN) on lipid synthesis of bovine hepatocytes, primary hepatocytes of calves were cultured in vitro and treated with different concentrations of INS and GLN: control group (0 nmol / L), concentration gradient group (1101001 000 nmol / L).After 1 hour culture, Western blotblotWB method was used, respectively.Fluorescence quantitative PCR qRT-PCR and immunofluorescence fluorescence assay to detect the expression of SREBP-1c mRNA in cultured hepatocytes treated with INS and GLN in vitro, the distribution of SREBP-1c nucleoplasm and the lipid metabolism of downstream target genes in hepatocytes treated with GLN and INS were determined.Effects of acetyl coA carboxylase (ACCs) and fatty acid synthase (FASN) on mRNA expression.The results showed that the expression level and transcription activity of SREBP-1c were significantly increased, the amount of nucleation was increased significantly, and the gene expression levels of ACC and FAS, the key enzymes of lipid synthesis in the downstream, were significantly increased by the treatment of 1: ins.On the contrary, GLN treatment significantly decreased the expression level and transcriptional activity of SREBP-1c, decreased the amount of nucleation, and the gene expression level of ACC and FAS, the key enzymes of lipid synthesis.These results suggest that ins can promote lipid synthesis and increase lipid deposition in hepatocytes by enhancing the activity of SREBP-1c, while GLN can decrease lipid synthesis in hepatocytes by inhibiting the activity of SREBP-1c.
【作者单位】：吉林大学动物医学学院;
【基金】：国家自然科学基金面上资助项目(31272621)
【分类号】：S858.23

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