为了阐明胰岛素(INS)、胰高血糖素(GLN)和瘦蛋白(LP)对牛脂肪细胞内激素敏感酯酶(HSL)、LP的mRNA表达的调控作用,本试验采用体外原代培养牛脂肪细胞的培养液中添加不同浓度的胰岛素(0、5、10、20、50、100mmol/L)、胰高血糖素(O、25、100、200、500、1000pg/mL)和瘦蛋白(O、2.5、5、10、50、100ng/mL),培养12h后,应用建立的实时荧光定量PCR(FQ-PCR)方法检测牛脂肪细胞内HSL、LP的mRNA丰度。结果表明:胰岛素对牛脂肪细胞内HSL的mRNA表达呈现剂量依赖性抑制作用,对LP的mRNA表达呈现剂量依赖性促进作用。胰高血糖素对牛脂肪细胞内HSL的mRNA表达呈现剂量依赖性促进作用,对牛脂肪细胞内LP的mRNA表达无作用。瘦蛋白对牛脂肪细胞内HSL的mRNA表达先呈现剂量依赖性促进作用,后呈现抑制作用。但对牛脂肪细胞内LP的mRNA表达呈现剂量依赖性抑制作用。
To clarify regulation of INS, GLN and LP on the mRNA expression of HSL and LP in vitro neonatal calf adipocytes, different concentrations of INS (0, 5, 10, 20, 50 and 100 mmol/L), GLN (0, 25, 100, 200, 500 and 1 000pg/mL),and LP (0, 2. 5, 5, 10, 50 and 100 ng/mL) were added respectively into media of primary monolayer culture calf adipocytes. After 12 h cultured, mRNA abundance of HSL and LP in the adipocytes was detected by FQ-PCR respectively. The results showed that INS plays a dose-dependently inhibitory role in abundance of HSL mRNA and a stimulating role in LP mRNA in calf adipocytes in vitro. However, action of GLN was contrary to INS in abundance of HSL mRNA, and plays a no role in abundance of LP mRNA in calf adipocytes in vitro. With concentrations of LP, abundance of HSL mRNA in calf adipocytes in vitro first dose-dependently increased, then decreased, but abundance of LP mRNA in calf adipocytes dose-dependently decreased in vitro.