摘要
取单层培养36h生长良好的犊牛肝细胞,采用单因素重复试验,分别添加0、2.5、5、10、50、100ng/mL的牛重组牛瘦蛋白(leptin),每个处理3个重复(每重复2孔),再培养12h后分别提取RNA和制备细胞上清液。应用荧光定量PCR方法检测外源NPY对肝细胞糖异生关键酶磷酸烯醇式丙酮酸羧激酶(Phosphoenolpyruvate carboxykinase,PEPCK)基因表达的影响,同时用比色法检测其对肝细胞PEPCK活性的影响。结果表明:一定浓度的leptin显著抑制了肝细胞PEPCK mRNA表达,降低了PEPCK活性。
A single factor duplicate test was designed to investigate whether leptin affects the expression of phosphoenolpyru- vate carboxykinase （PEPCK） mRNA and the activity of PEPCK in vitro cultured bovine hepatocyte. Leptin was added to the media with 0,2.5,5,10,50,100 ng/mL. Abundance of PEPCK mRNA in vitro cultured bovine hepatocyte cultured for 12 hours after leptin was added was determined by real-time fluorescence quantitative RT-PCR, and the activity of PEPCK was determined by colourimetry. The results showed that the leptin could depress the expression of PEPCK mRNA and lower the activity of PEPCK in vitro cultured bovine hepatocyte.
出处
《中国草食动物》
CAS
2008年第1期6-8,共3页
China Herbivores
基金
国家自然科学基金重点项目(30230260)
