【摘要】 研究新近发现的猪Ghrelin(porcine ghrelin,pGhrelin)对猪前体脂肪细胞Caspase-3活性及其基因表达的影响.采用细胞培养技术,以仔猪背部皮下前体脂肪细胞为靶细胞,经0、1、10和100nmol/L pGhrelin处理细胞48h后,于倒置生物显微镜下进行脂肪细胞形态学观察.利用MTT法测定pGhrelin对细胞增殖的影响.采用分光光度法检测Caspase-3活性.以实时荧光定量RT-PCR方法测定Caspase-3的基因表达.结果显示,10nmol/L pGhrelin可以显著降低脂肪细胞Caspase-3的活性与mRNA的表达水平(P<0.05),100nmol/L pGhrelin对猪脂肪前体细胞增殖有极显著促进作用(P<0.01).上述结果表明,pGhrelin可以下调Caspase-3的活性与基因表达,促进脂肪细胞增殖,抑制脂肪细胞凋亡,其机制可能与Caspase-3依赖性凋亡调节信号通路有关.更多还原

【Abstract】 To investigate the effects of porcine Ghrelin(pGhrelin), a newly discovered peptide on porcine preadipocyte Caspase-3 activity and gene expression, piglet subcutaneous preadipocytes treated with 0, 1, 10 and 100 nmol/L pGhrelin were cultured for 48 h, and preadipocyte morphology was observed under an inverted biological microscope, the effects of preadipocyte prolifaration was mesured by MTT, preadipocyte Caspase-3 activity was detected with spectrophotometric method, Caspase-3 gene expression level was determined using real-time fluorescent quantitative RT-PCR.10 nmol/L pGhrelin significantly decreased preadipocyte Caspase-3 activity and mRNA expression level(P < 0.05), 100 nmol/L pGhrelin significantly promoted porcine preadipocyte proliferation compared to control group(P < 0.01).pGhrelin downregulated Caspase-3 activity and expression mRNA of porcine preadipocyte, induced preadipocyte proliferation and inhibited apoptosis.Caspase-3 dependent apoptosis regulation signalling was supposed to be involved in the apoptosis mechanism.更多还原