摘要
目的观察胎球蛋白-a(fetuin-a)对脂肪分解的影响并探讨其可能的机制。方法体外培养3T3-L1前脂肪细胞,分化成熟后用不同浓度的fetuin-a孵育。采用甘油检测试剂盒测定释放到上清液的甘油含量;采用Real-timePCR检测细胞中脂肪甘油三酯脂肪酶(Adipose triglyceride lipase,ATGL)的mRNA表达;采用western blot检测细胞内磷酸化激素敏感脂肪酶(Hormone Sensitive Lipase,HSL)和ATGL的蛋白表达;采用ELISA试剂盒检测细胞内环磷酸腺苷(cyclic adenosine monophosphate,cAMP)的含量。结果 Fetuin-a能够抑制成熟脂肪细胞的脂肪分解,具有剂量依赖关系。Fetuin-a能够降低细胞内cAMP及磷酸化的HSL含量,同时使脂肪细胞ATGL mRNA水平及蛋白表达降低。结论 Fetuin-a抑制3T3-L1脂肪细胞的脂肪分解,其机制可能是通过抑制cAMP-PKA-HSL通路以及降低细胞内ATGL的表达而发挥作用的。
Objective TO observe the effect of fetuin-a on lipolysis of 3T3-L1 adipocytes and to explore its regulation mechanism. Methods 3T3-L1 preadipocytes were induced under standard differentiation process and treated with different concentration of fetuin-a for 12h after differentiating. Lipolysis of adipoeytes was examined by the measurement of glycerol release; The expressions of mRNA of ATGL were analysed using Real-time PCR;The expressions of protein of phospho-HSL,ATGL were analysed using western blot. cAMP levels in cell extracts were measured using a ELISA kit. Results Fetuin-a inhibited basal lipolysis in adipocytes in a dose-dependent manner. Fetuin-a decreased intraeellular cAMP production and the expression of phospho-HSL protein. Fetuin-a also decreased the expressions of ATGL mRNA and protein. Conclusion These results suggested that fetuin a inhibited lipolysis of adipocyte through inactivation of signaling cascades mediated by cAMP-PKA-HSL and ATGL.
出处
《中国实验诊断学》
2012年第6期965-968,共4页
Chinese Journal of Laboratory Diagnosis
基金
上海市科委资助项目(034119816)
