全反式视黄酸在牛前脂肪细胞增殖及分化中的作用  

Effects of all-trans retinoic acid on cell proliferation and differentiation in bovine preadipocytes

作　　者：范云珲 逯璐 许秋实[1] 郑晰丹 杨童 徐闯[1] FAN Yunhun;LU Lu;XU Qiushi;ZHENG Xidan;YANG Tong;XU Chuang(College of Animal Science and Technology,Heilongjiang Bayi Agricultural University,Daqing,Heilongjiang 163000,China)

机构地区：[1]黑龙江八一农垦大学动物科技学院,黑龙江大庆163000

出　　处：《中国兽医学报》2022年第11期2256-2260,2291,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(32072931)。

摘　　要：奶牛前脂肪细胞的增殖与分化对于正常脂肪组织发育至关重要。本研究旨在细胞水平探讨全反式视黄酸(ATRA)对牛前脂肪细胞的增殖与分化。试验分为2组,分别是试验组(牛前脂肪细胞用20 nmol/L ATRA处理48 h)及对照组。用MTS及EdU及Ki67法检测ATRA对牛前脂肪细胞增殖的影响,免疫蛋白印记(Western blot)检测ATRA对牛前脂肪细胞中几种增殖相关蛋白表达的影响,包括细胞周期蛋白D1(CCND1)、2、3和CCNE1及细胞周期蛋白激酶(CDK2、4、6);使用流式细胞术检测细胞周期进程,并检测分化相关指标PPARγ和C/EBPα蛋白表达情况。结果显示,与对照组相比,20 nmol/L ATRA可抑制牛前脂肪细胞增殖,同时降低S期细胞比例并降低细胞周期蛋白(CCND1、CCND2、CCND3和CCNE1)和细胞周期蛋白依赖性激酶(CDK2、CDK4和CDK6)的蛋白丰度。此外,与对照组相比,ATRA的添加还通过下调PPARγ和C/EBPα的蛋白质表达来抑制牛前脂肪细胞的分化。结果表明,20 nmol/L ATRA处理48 h会抑制牛前脂肪细胞的增殖与分化,对正常脂肪组织发育具有潜在作用。Proliferation and differentiation of preadipocytes in dairy cows are essential for the development of adipose tissue.The aim of the present study was to investigate the proliferation and differentiation of all-trans retinoic acid(ATRA)in a wide variety of cells,including bovine preadipocytes at the cellular level.The bovine preadipocytes of the experimental group were treated with 20 nmol/L ATRA for 48 h and the control group was left untreated.The effect of ATRA on the proliferation of bovine preadipocytes was determined by the MTS,EdU and Ki67 assays,the effect of ATRA on the expression of several proliferation-related proteins including cyclin D1(CCND1),cyclin D2(CCND2),cyclin D3(CCND3),CCNE1 and cyclin-dependent kinases(CDK2,4,6)in bovine preadipocytes was determined by Western blot.Cell cycle progression was examined by flow cytometry analysis and protein expression of differentiation-related indicators(PPARγand C/EBPα)were detected the by Western blot.The results showed that bovine preadipocytes proliferation was inhibited after ATRA(20 nmol/L)treatment compared with the control group.Meanwhile,the proportion of cells in the S phase was increased,protein abundance of cyclins(CCND1,CCND2,CCND3 and CCNE1)and cyclin-dependent kinases(CDK2,CDK4 and CDK6)were decreased.In addition,ATRA treatment inhibited the differentiation of bovine preadipocytes by the downregulation of PPARγand C/EBPα.In summary,these results indicate that treatment with 20 nmol/L ATRA inhibits the proliferation and differentiation of bovine preadipocytes after 48 h,which has a potential effect on the development of normal adipose tissue.

关 键 词：ATRA 增殖 分化 牛前脂肪细胞 

分 类 号：S856.5[农业科学—临床兽医学]