College of Yingdong agricultural science and engineering, Shaoguan university, Daxue road, Zhenjiang district, Shaoguan 512005, China.
College of Yingdong agricultural science and engineering, Shaoguan university, Daxue road, Zhenjiang district, Shaoguan 512005, China.
Ann Endocrinol (Paris). 2014 Feb;75(1):1-9. doi: 10.1016/j.ando.2013.10.003. Epub 2014 Feb 16.
Obestatin, originally identified and purified from rat stomach extracts, was reported to bind to orphan G protein-coupled receptor, GPR39, and inhibit appetite and gastric motility. This study was conducted to investigate the effects of porcine obestatin on proliferation, differentiation and apoptosis of porcine preadipocytes isolated from subcutaneous fat of piglets. At indicated times of culture, morphology of preadipocytes and accumulated lipid droplets within the cells were identified by invert microscope. After treating with obestatin (0, 0.1, 1, 10 and 100nM), cell proliferation was measured by MTT method and protein expression of CCAAT/enhancer binding protein-α (C/EBPα), peroxisome proliferator-activated receptor-γ (PPARγ), Caspase-7 and Caspase-9 was determined by Western Blot, mRNA expression of GPR39 and Caspase-3 was analyzed by RT-PCR, and the activity of Caspase-3 was measured by spectrophotometric method. The results showed that obestatin had no effect on GPR39 expression, while promotes the optical density (OD) value of cells, enhanced protein expression of PPARγ and C/EBPa, decreased mRNA expression and activity of Caspase-3, and inhibited protein expression of Caspase-7 and Caspase-9 in a dose-dependent manner. These results suggested that obestatin enhances proliferation and differentiation of preadipocytes promoting PPARγ and C/EBPa expression, and inhibiting preadipocyte apoptosis by decreasing expression of Caspase-3, Caspase-7 and Caspase-9.
肥胖抑制素最初从大鼠胃提取物中被鉴定和纯化，被报道与孤儿 G 蛋白偶联受体 GPR39 结合，并抑制食欲和胃动力。本研究旨在探讨猪肥胖抑制素对从小猪皮下脂肪分离的猪前体脂肪细胞增殖、分化和凋亡的影响。在培养的指定时间，通过倒置显微镜鉴定前体脂肪细胞的形态和细胞内积累的脂滴。用肥胖抑制素（0、0.1、1、10 和 100nM）处理后，通过 MTT 法测量细胞增殖，通过 Western Blot 法测定 CCAAT/增强子结合蛋白-α（C/EBPα）、过氧化物酶体增殖物激活受体-γ（PPARγ）、Caspase-7 和 Caspase-9 的蛋白表达，通过 RT-PCR 分析 GPR39 和 Caspase-3 的 mRNA 表达，通过分光光度法测定 Caspase-3 的活性。结果表明，肥胖抑制素对 GPR39 表达没有影响，而促进细胞光密度（OD）值增加，增强 PPARγ 和 C/EBPα 的蛋白表达，降低 Caspase-3 的 mRNA 表达和活性，并呈剂量依赖性抑制 Caspase-7 和 Caspase-9 的蛋白表达。这些结果表明，肥胖抑制素通过降低 Caspase-3、Caspase-7 和 Caspase-9 的表达，促进 PPARγ 和 C/EBPα 的表达，增强前体脂肪细胞的增殖和分化，抑制前体脂肪细胞凋亡。
