1,25(OH)_(2)D_(3)以双向方式影响猪前体脂肪细胞增殖分化的研究  

1,25(OH)_(2)D_(3) Affects Proliferation and Differentiation of Porcine Preadipocytes in a Biphasic Manner

作　　者：岳小婧 张晶 马姝雯[1] 曹忻 卢建雄[1] 张国华[1] YUE Xiaojing;ZHANG Jing;MA Shuwen;CAO Xin;LU Jianxiong;ZHANG Guohua(College of Life Science and Engineering,Northwest Minzu University,Lanzhou 730030,China)

机构地区：[1]西北民族大学生命科学与工程学院,兰州730030

出　　处：《中国畜牧兽医》2021年第7期2349-2357,共9页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金项目(31860633);国家民委中青年英才计划([2018]98);甘肃省自然科学基金项目(20JR5RA504);西北民族大学中央高校基本科研业务费创新团队项目(31920190001);西北民族大学研究生科研创新项目(Yxm2020129)。

摘　　要：1,25(OH)_(2)D_(3)是维生素D的主要活性形式,影响人和动物脂肪形成,为探究其在猪脂肪细胞增殖分化中的作用,试验从3~5日龄仔猪皮下脂肪组织分离培养前体脂肪细胞,并以浓度为0、0.1、1、10、100和1000 nmol/L的1,25(OH)_(2)D_(3)分别处理,在培养的0、1、2、4、6、8和10 d采用MTT比色法检测细胞增殖活性;在诱导分化后0、1、2、4、6、8和10 d,以油红O染色提取法和实时荧光定量PCR检测细胞成脂分化及分化标志基因过氧化物酶体增殖物激活受体γ(PPARγ)和脂肪酸合成酶(FAS)表达。结果显示,0.1和1 nmol/L 1,25(OH)_(2)D_(3)显著促进猪前体脂肪细胞增殖(P<0.05),而浓度为10~100 nmol/L时则抑制细胞增殖(P<0.05);0.1和1 nmol/L 1,25(OH)_(2)D_(3)显著抑制猪前体脂肪细胞分化(P<0.05),降低PPARγ和FAS mRNA表达水平(P<0.05),但在浓度为10和100 nmol/L时,显著促进猪前体脂肪细胞分化(P<0.05),上调PPARγ和FAS mRNA表达(P<0.05);浓度达1000 nmol/L时,可能对细胞有毒性作用。综合以上结果,低浓度1,25(OH)_(2)D_(3)促进猪前体脂肪细胞增殖,而通过下调PPARγ表达抑制分化;高浓度1,25(OH)_(2)D_(3)抑制猪前体脂肪细胞增殖,通过上调PPARγ表达促进分化。1,25(OH)_(2)D_(3)对猪前体脂肪细胞增殖和分化具有双向作用。As the main active form of vitamin D,1,25(OH)_(2)D_(3) affects adipogenesis of humans and animals,but its role in proliferation and differentiation of porcine adipocytes is unclear.To explore the effects of 1,25(OH)_(2)D_(3) on proliferation and differentiation of porcine preadipocytes,the cells isolated from subcutaneous adipose tissue of 3-5 days piglets were cultured and treated with 0,0.1,1,10,100 and 1000 nmol/L concentration of 1,25(OH)_(2)D_(3),respectively.The proliferation of preadipocytes cultured for 0,1,2,4,6,8 and 10 d was detected by MTT method.The differentiation and expression levels of PPARγand FAS mRNA for the preadipocytes at 0,1,2,4,6,8 and 10 d after inducing-differentiation were determined by oil red O staining and Real-time quantitative PCR,respectively.The results showed that 0.1 and 1 nmol/L 1,25(OH)_(2)D_(3) significantly promoted proliferation of porcine preadipocytes(P<0.05),but the concentration of 10-100 nmol/L inhibited the cell proliferation(P<0.05).At the concentration of 0.1 and 1 nmol/L,1,25(OH)_(2)D_(3) markedly inhibited the differentiation of porcine preadipocytes and reduced mRNA expression of PPARγand FAS(P<0.05).On the contrary,10 and 100 nmol/L 1,25(OH)_(2)D_(3) significantly enhanced differentiation of porcine preadipocytes and increased mRNA expression of PPARγand FAS(P<0.05).When the concentration was 1000 nmol/L,it might be toxic to the cells.These results suggested that low concentration of 1,25(OH)_(2)D_(3) could promote the proliferation of porcine preadipocytes,but inhibit differentiation through down-regulating PPARγexpression.High concentration of 1,25(OH)_(2)D_(3) could inhibit the proliferation of the preadipocytes and promote differentiation by up-regulating PPARγexpression.And so,1,25(OH)_(2)D_(3) affected proliferation and differentiation of porcine preadipocytes in a biphasic manner.

关 键 词：1 25(OH)_(2)D_(3) 猪 前体脂肪细胞 增殖 分化 

分 类 号：Q253[生物学—细胞生物学]