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  

Effects of insulin and glucagon on the lipid synthesis in primary cultured bovine hepatocytes in vitro

作　　者：张世奇[1] 丁红研[1] 史珍[1] 张强[1] 李心慰[1] 李小兵[1] 

机构地区：[1]吉林大学动物医学学院,吉林长春130062

出　　处：《中国兽医学报》2017年第2期312-317,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金面上资助项目(31272621)

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。To determine the effect of insulin and glucagon on the hepatic lipid synthesis in dairy cattle,bovine hepatocytes were cultured and treated with different concentrations of insulin （INS） andglucagon （GLN） with 0,1,10,100 and 1 000 nmol/L. After treated for 1 h,Western blot （WB） method, fluorescence quantitative PCR （qRT-PCR） method and cell immunofluorescence （IF） method were respectively used to detect the expression of key transcription factors such as sterol regulatory element binding protein lc （SREBPqc） and its target genes such as acetyl-CoA carboxylase （ACC） and fatty acid synthase （FAS）. The results showed that INS significantly increased the expression and transcriptional activity of SREBP-1c, and markedly increased the expression levels of ACC and FAS gene. Oppositely,GLN significantly decreased the SREBP-1c expression and transcriptional activity, and significantly decreased the ACC and FAS gene expression. These results indicated that INS could activate the SREBP-1c and then promote liver cell lipid synthesis and increase liver fat deposition. Nevertheless,GLN could inhibit SREBP-1c and decrease the liver cell lipid synthesis.

关 键 词：胰岛素 胰高血糖素 固醇调节元件结合蛋白-1C 犊牛肝细胞 脂合成 

分 类 号：S856.5[农业科学—临床兽医学]