【摘要】 体外原代培养犊牛肝细胞,添加不同浓度的胰岛素(insulin,INS)(0、1、10、100、1 000nmol/L)和胰高血糖素(glucagon,GLN)(0、1、10、100、1 000nmol/L),每个处理做3个重复,分别培养1h后,提取总RNA。实时荧光定量PCR(qRT-PCR)的方法,检测肉碱棕榈酰转移酶Ⅰ(carnitine palmitoyl transferase,CPT-Ⅰ)和肉碱棕榈酸转移酶Ⅱ(carnitine palmitoyl transferase,CPT-Ⅱ)mRNA表达的变化。结果显示,随着培养液中INS含量的升高,肝细胞中CPT-ⅠmRNA和CPT-ⅡmRNA的表达量逐渐降低,呈现一定的剂量依赖性。随着培养液中GLN含量的升高,肝细胞中CPT-ⅠmRNA和CPT-ⅡmRNA的表达量均升高,呈现一定的剂量依赖性,且当GLN的添加浓度高于100nmol/L时,CPT-ⅠmRNA和CPT-ⅡmRNA的表达量显著升高(P<0.01)。结果表明,INS可在一定程度上抑制脂氧化关键酶的表达,GLN可显著促进脂氧化关键酶的表达。更多还原

【Abstract】 INS and GLN were added to the media with the final concentrations of 0,1,10,100, 1 000nmol/L,respectively.After 1-hour culture,hepatocytes were collected and total RNA was extracted.The expression levels of carnitine palmitoyl transferase-Ⅰ(CPT-Ⅰ)and carnitine palmitoyl transferase-Ⅱ(CPT-Ⅱ)mRNA were determined by using fluorescent quantitative real time PCR(qRT-PCR).The present results showed that the expression levels of CPT-Ⅰand CPT-Ⅱ decreased in a dose-dependent manner after 1hINS-treated compared with untreated(UT) groups,but there was no statistical significance.The expression levels of CPT-Ⅰand CPT-Ⅱincreased in a dose-dependent manner after 1hGLN-treated when gradually increased glucagon in the culture media and were significantly higher when the concentrations of GLN added in the media exceed 100nmol/L.In conclusion,the results of this study demonstrated that the expression of the key enzymes of lipid oxidation were inhibited by INS and were promoted significantly by GLN.更多还原