CAJ | 학술논문

取单层培养72 h生长良好的犊牛肝细胞,采用单因素重复试验,分别添加0、25、50、100、200、400 ng/L的牛重组抵抗素(resistin),每个处理3个重复(每重复2孔).继续培养12 h后分别提取RNA并制备细胞上清液.应用荧光定量PCR方法检测牛重组Resistin对肝细胞糖异生关键酶丙酮酸羧化酶(Pyruvate carboxylase,PC)基因表达的影响,同时用比色法检测其对肝细胞PC酶活性的影响.结果表明,一定浓度的resistin显著下调了肝细胞PCmRNA表达,且降低了PC酶活性.
취단층배양72 h생장량호적독우간세포,채용단인소중복시험,분별첨가0、25、50、100、200、400 ng/L적우중조저항소(resistin),매개처리3개중복(매중복2공).계속배양12 h후분별제취RNA병제비세포상청액.응용형광정량PCR방법검측우중조Resistin대간세포당이생관건매병동산최화매(Pyruvate carboxylase,PC)기인표체적영향,동시용비색법검측기대간세포PC매활성적영향.결과표명,일정농도적resistin현저하조료간세포PCmRNA표체,차강저료PC매활성.
A single factor duplicate test was designed to investigate whether bovine recombinded resistin impacts the expression of pyruvate carboxylase (PC) mRNA and the activity of PC in vitro culture bovine hepatocyte.Bovine recombinded resistin was added to the media with 0,25,50,100,200 and 400 ng/L.Abundance of PC mRNA in bovine hepatocyte,which was cultured with bovine recombinded resistin for 12 hours,was determined by real-time fluorescence quantitative RT-PCR,and activity of PC was determined by colourimetry.The results showed that bovine recombinded resistin could downregulate the expression of PC mRNA and the activity of PC in vitro culture bovine hepatocyte.