中国农业科学Issue(18)：3878-3886,9.DOI:10.3864/j.issn.0578-1752.2013.18.018
烟酰胺削弱罗格列酮诱导猪前体脂肪细胞凋亡的作用与分子机制
Effects and Molecular Mechanism of Nicotinamide on Attenuating Apoptosis of Rosiglitazone-induced Porcine Preadipocytes
摘要
Abstract
[Objective]This study was performed to explore the effect and molecular mechanism of nicotinamide (NIC) and rosiglitazone (RSG) on apoptosis of porcine preadipocytes for finding a novel method to control adipogenesis through regulation of preadipocyte apoptosis.[Method]Cells were cultured in high glucose DMEM medium containing 0, 0.2, 0.4, 0.6, 0.8 and 1.0 mmol·L-1 RSG. After being treated for 48 h, cells were stained with Hoechst33258 and Annexin-V-FITC/PI and observed under microscope. Simultaneously, cells were harvested for analysis of flow cytometry and collection of total protein to detect protein levels of several genes related with apoptosis. Based on the above results, cells were further co-cultured in DMEM medium containing 0, 0.1, 0.2 and 0.3 mmol·L-1 NIN with 1 mmol·L-1 RSG. After being treated for 48 h, cells were stained and observed. Cells were harvested for analysis of flow cytometry and collection of total, cytoplasmic and nuclear proteins for Western blotting analysis.[Result]RSG induced preadipocyte apoptosis by a dose-dependent way and reduced cell number. The levels of Bcl-2 and cleaved Caspase-3 increased, whereas the level of Bax decreased in higher concentration of RSG. Moreover, 0.1(P<0.05)、0.2(P<0.01)and 0.3 mmol·L-1 (P<0.01)NIC significantly attenuated RSG-induced apoptosis of preadipocytes. The levels of Bcl-2 and cleaved Caspase-3 were down-regulated, whereas the level of Bax was up-regulated in treatment with 0.3 mmol·L-1 NIN + 1 mmol·L-1 RSG. Compared with the control (1 mmol·L-1 RSG+0 mmol·L-1 NIC), in treatment with 0.3 mmol·L-1 NIC+1 mmol·L-1 RSG, Sirt1 was transfered from cytoplasm into nucleus and the level of cleaved Caspase-3 was reduced.[Conclusion]NIC attenuated RSG-induced apoptosis of porcine preadipocytes via cytoplasm/nucleus transposition of Sirt1 and down-regulating the level of nuclear cleaved Caspase-3.关键词
猪/烟酰胺/罗格列酮/前体脂肪细胞/凋亡Key words
pig/nicotinamide/rosiglitazone/preadipocyte/apoptosis引用本文复制引用
庞卫军,许儒祥,熊燕,孙世铎,杨公社..烟酰胺削弱罗格列酮诱导猪前体脂肪细胞凋亡的作用与分子机制[J].中国农业科学,2013,(18):3878-3886,9.基金项目
国家自然科学基金项目（30600437）、国家“973”计划项目（2012CB124705）、西北农林科技大学基本科研业务项目（QN2009021）和国家生猪产业技术体系（CARS-36）和陕西省生猪产业技术体系 （）
