RHIGF_对体外单层培养的肝细胞PEPCKmRNA丰度的影响.pdfVIP

434 2008 4 28 4 Chin J Vet Sci Apr. 2008 Vol. 28 o. 4 RHIGF PEPCK mRNA 1 1 1* 2 2 2 邓俊良, 王 娅 , 崔恒敏 , 徐 闯, 刘国文 , 王 哲 ( 1. , 625014; 2. , 130062) :根据牛肝细胞中持家基因表达的恒定性, 选 肌动蛋白(actin) 作为内参照, 检测外源性添加不同质量浓 度的重组人胰岛素样生长因子(RHIGF) 对体外培养牛肝细胞磷酸烯醇式丙酮酸羧激酶( PEPCK)mR A 表达水平 的变化情况选 体外培养的犊牛单层肝细胞为实验模型, 当细胞贴壁后在无血清培养液中添加RHIGF ,其终 质量浓度分别为0510152030405060100150g/L,待添加8 h 后, 提取总 R A, 反转录, 相同的模板扩增 PEPCK基因及 内参, 再比较各梯度目的基因与内参的灰度结果表明, 随着添加 RHIGF 质量浓度的升高, PEPCK mR A 表达呈下降趋势,且均低于对照组 : 内源性竞争PCR;RHIGF ;PEPCK mR A 丰度; 肝细胞 :S852.23 :A :2008) Regulation of the expression of the phosphoenolpyruvate carboxy inase gene in cultured bovine hepatocytes by recombination human insulinli e growth factor 1 1 1* 2 2 2 DE G Junliang ,WA G Ya ,CUI Hengmin ,XU Chuang ,LIU Guowen ,WA G Zhe ( 1. College of Veteri nary Medicine, Sichuan Agricultural University, Ya an, Sichuan 625014, China; 2. College of Animal Science and Veterinary Medicine, Jilin University , Changchun 130062, China) Abstract:The depression effect of phosphoenolpyruvate carboxykinase(PEPCK) by recombination human insulinlike growth factor (RHIGF )was studied in primary rat hepatocyte cultures by detection the expression level of actin mR A.RHIGF was added to the serum freemedium as various concentration of 0,5, 10, 15,20,30,40,50,60, 100, 150 g/L in adherence cell.8 h later,total R A was extracted and reversed transcript, amplify the PEPCK gene and actin gene according to the same template, compared to all the gradient of the two gene.The result indicated that the mR A level of PEPCK in hepatocytewas kept tendency of downwards after addition of RHIGF . Key words: endogenous competitive PCR;RHIGF ; expression of PEPCK mR A;hepatocytes *