Wang Pei H, Ko Ya H, Chin Hong J, Hsu Chichen, Ding S T, Chen Ching Y
Department of Animal Science and Technology/Center for Biotechnology, National Taiwan University, Taipei 106, Taiwan.
Comp Biochem Physiol B Biochem Mol Biol. 2009 Aug;153(4):327-31. doi: 10.1016/j.cbpb.2009.04.003. Epub 2009 Apr 19.
To study the role of sterol regulatory element-binding proteins (SREBP) in lipogenesis and cholesterol synthesis in the chicken, two experiments were carried out. In the first study, seven-week-old broilers (n=16) were allocated into 2 groups, fasted for 24 h or refed for 5 h after a 24 h fasting. The mRNA concentrations for SREBPs and other lipogenic genes in the liver were determined by quantitative real time PCR. The hepatic mRNA relative abundance of lipogenic genes and genes involved in cholesterol synthesis were significantly greater (p<0.001) in the refed broilers. Similar results were demonstrated with Northern analysis. The data suggest that in the liver of fasted broilers, genes associated with lipogenesis and cholesterol biosynthesis were inhibited. Indeed, the mRNA concentrations for fatty acid synthase (FAS), malic enzyme, and stearoyl coenzyme A desaturase were almost undetectable after the 24 h fasting. The data also demonstrated that the expression of lipogenic genes coordinate well as a group during the refeeding period. Second, three small interfering RNA (siRNA) oligonucleotides against SREBP1 were designed to be used in transfecting a chicken hepatocarcinoma cell line LMH. One of the three siRNAs effectively reduced SREBP1 mRNA concentration (p<0.01). The acetyl coenzyme A carboxylase(alpha) (ACC(alpha)) mRNA was also significantly reduced by the SREBP1 siRNA treatment, suggesting that SREBP1 can upregulate the expression of this lipogenic gene. This siRNA, however, did not affect the mRNA for FAS. Taken together, the RNA interference study showed that SREBP1 has the ability to regulate the expression of ACC(alpha). This study has helped us understand more about the function of SREBP1 and the physiology of the broiler chickens.
为研究固醇调节元件结合蛋白(SREBP)在鸡脂肪生成和胆固醇合成中的作用,进行了两项实验。在第一项研究中,将7周龄的肉鸡(n = 16)分为2组,一组禁食24小时,另一组在禁食24小时后再喂食5小时。通过定量实时PCR测定肝脏中SREBP和其他脂肪生成基因的mRNA浓度。再喂食的肉鸡肝脏中脂肪生成基因和参与胆固醇合成的基因的mRNA相对丰度显著更高(p<0.001)。Northern分析也得到了类似结果。数据表明,在禁食肉鸡的肝脏中,与脂肪生成和胆固醇生物合成相关的基因受到抑制。事实上,禁食24小时后,脂肪酸合酶(FAS)、苹果酸酶和硬脂酰辅酶A去饱和酶的mRNA浓度几乎检测不到。数据还表明,在再喂食期间,脂肪生成基因作为一个整体协调良好地表达。其次,设计了三种针对SREBP1的小干扰RNA(siRNA)寡核苷酸,用于转染鸡肝癌细胞系LMH。三种siRNA中的一种有效地降低了SREBP1 mRNA浓度(p<0.01)。SREBP1 siRNA处理也显著降低了乙酰辅酶A羧化酶(α)(ACC(α))mRNA,表明SREBP1可以上调这种脂肪生成基因的表达。然而,这种siRNA不影响FAS的mRNA。综上所述,RNA干扰研究表明SREBP1有能力调节ACC(α)的表达。这项研究有助于我们更多地了解SREBP1的功能和肉鸡的生理学。