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首页> 外文期刊>Molecular and Cellular Biochemistry >Expression of TGH and its role in porcine primary adipocyte lipolysis
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Expression of TGH and its role in porcine primary adipocyte lipolysis

机译：TGH的表达及其在猪原代脂肪细胞脂解中的作用

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Aims Triacylglycerol hydrolase (TGH) plays an important role in intracellular lipid metabolism. However, most previous studies on TGH were focused on rodent animals. Pig is considered as one of the best models for studying obesity, diabetes, and other lipid metabolism related diseases. So far, we do not know whether TGH is to play a role in lipolysis in porcine primary adipocyte through stimulation by hormones as Hormone-sensitive lipase (HSL). The objective of this study is to explore the mechanisms that regulate TGH expression in porcine adipocyte and its role in fasting-induced and Isoproterenol-stimulated lipolysis in porcine primary adipocytes. Materials and methods Stromal-vascular cells containing preadipocytes were isolated from cervical and dorsal subcutaneous adipose tissues of approximately 3-day-old Chinese male piglets. After confluence, the differentiation was induced by Insulin, hydrocortisone, and transferrin. The primary adipocytes were cultured in various concentration (0.1 to 200 μmol/l) Isoproterenol for 0–12 h or serum-free medium after differentiation. The glycerol release and triacylglycerol (TG) contents were detected when the cells were collected. Then, expression of TGH and HSL mRNA and their protein were determined by RT-PCR, Western Blot, and lipolytic analysis. Results Both TGH mRNA and protein were not detected on day 0, but as differentiation was induced, their expression displayed a great increase. The expression of TGH mRNA showed the highest level on day 4, but its protein reached the highest level on day 6 and began to fall down from day 8. The expression of TGH mRNA and proteins was increased in serum-free media, and mRNA expression was decreased while changed into complete media again. The glycerol release increased significantly when the cells were cultured with serum-free media. The lower concentration of Isoproterenol (0.1 and 1 μmol/l) did not affect the expression of TGH and HSL, but higher concentration (10, 100, and 200 μmol/l) could greatly up-regulate HSL expression but did not affect TGH level. Also, the higher concentration of Isoproterenol could increase the glycerol release and decrease TG content in dose-dependent manner. Conclusions These results suggested that TGH expression is differentiation-dependent in porcine primary adipocytes and TGH plays a role in fasting-induced lipolysis not hormone-stimulated lipolysis.
机译：目的三酰基甘油水解酶（TGH）在细胞内脂质代谢中起重要作用。但是，以前有关TGH的大多数研究都集中在啮齿动物上。猪被认为是研究肥胖，糖尿病和其他脂质代谢相关疾病的最佳模型之一。到目前为止，我们还不知道TGH是否通过激素作为激素敏感性脂肪酶（HSL）的刺激在猪原代脂肪细胞的脂解中发挥作用。这项研究的目的是探索调节猪脂肪细胞中TGH表达的机制及其在猪原代脂肪细胞中由空腹诱导和异丙肾上腺素刺激的脂解中的作用。材料和方法从约3天大的中国雄性仔猪的子宫颈和背部皮下脂肪组织中分离出含有前脂肪细胞的基质血管细胞。汇合后，分化是由胰岛素，氢化可的松和转铁蛋白诱导的。分化后，将原代脂肪细胞以各种浓度（0.1至200μmol/ l）异丙肾上腺素或无血清培养基培养0-12小时。当收集细胞时，检测到甘油释放和三酰基甘油（TG）含量。然后，通过RT-PCR，蛋白质印迹和脂解分析确定TGH和HSL mRNA及其蛋白的表达。结果在第0天均未检测到TGH mRNA和蛋白，但随着分化的诱导，其表达量显着增加。 TGH mRNA的表达在第4天达到最高水平，但其蛋白在第6天达到最高水平，并从第8天开始下降。在无血清培养基中TGH mRNA和蛋白的表达增加，并且mRNA表达被减少，而再次更改为完全媒体。当用无血清培养基培养细胞时，甘油的释放显着增加。较低浓度的异丙肾上腺素（0.1和1μmol/ l）不会影响TGH和HSL的表达，但是较高的浓度（10、100和200μmol/ l）可以大大上调HSL的表达，但不影响TGH的水平。同样，较高浓度的异丙肾上腺素可以剂量依赖性方式增加甘油的释放并降低TG含量。结论这些结果表明，TGH在猪原代脂肪细胞中表达是依赖于依赖性的，TGH在空腹诱导的脂肪分解而不是激素刺激的脂肪分解中起作用。

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