PTEN过表达对奶牛肝细胞脂肪酸氧化代谢的影响

Effect of PTEN overexpression on fatty acid oxidation in hepatocytes of cows

目的探讨PTEN过表达对奶牛肝细胞脂肪酸氧化代谢的影响。方法构建PTEN过表达的重组腺病毒载体,病毒扩增后,感染犊牛原代肝细胞,并设空白对照组(加入PBS)和阴性对照组(感染阴性对照腺病毒AD-GFP),感染48 h后,荧光定量PCR检测细胞中PTEN、脂肪酸氧化代谢转运主要基因CPTⅠ、CPTⅡm RNA的表达水平,Western blot检测相应蛋白表达水平。结果重组腺病毒载体经酶切鉴定证明构建正确。过表达PTEN基因后,可升高CPTⅠ和CPTⅡ基因m RNA和蛋白的表达量(P<0.01)。结论 PTEN通过调节脂肪酸氧化酶转录和翻译水平,对脂肪酸在肝细胞内的氧化代谢过程进行调节,其有望作为临床有效的生物调控药物,用于治疗奶牛脂肪肝。

Objective To investigate the effect of PTEN overexpression on fatty acid oxidation in hepatocytes of cows.Methods The recombinant adenovirus vector for PTEN overexpression was constructed,and the amplified virus was infected to primary hepatocytes of newborn calf,using PBS as blank control and AD-GFP as negative control.The transcription levels of PTEN,carnitine palmityl transferase（CPT）Ⅰ and CPT Ⅱ 48 h after infection were determined by fluorescent quantitative PCR,while the protein expression levels by Western blot.Results Restriction analysis proved that recombinant adenovirus vector was constructed correctly.The PETN overexpression increased the expression levels of m RNAs and proteins of CPTⅠand CPTⅡ（P 0.01）.Conclusion PTEN regulated the fatty acid oxidation in hepatocytes by regulating the transcription and translation levels of fatty acid oxidase,which might be used as an effective biological regulating agent for treatment of fatty liver in clinic.