多不饱和脂肪酸对小尾寒羊前体脂肪细胞增殖的影响  

Effects of polyunsaturated fatty acids on the proliferation of preadipocytes in Small Tail Han sheep

作　　者：卢美琳 阿尔祖古丽 李自良 敖慧娟 乔自林[1,3] 阿依木古丽[1,2] LU Meilin;Aerzuguli;LI Ziliang;AO Huijuan;QIAO Zilin;Ayimuguli(Gansu Tech Innovation Center of Animal Cell,Biomedical Research Center,Northwest Minzu University,Lanzhou 730030,China;Science and Engineering College of Northwest University for Nationalities,Lanzhou 730030,China;Northwest University for Nationalities Key Laboratory of Bioengineering and Technology,National Civil Committee,Biomedical Research Center,Lanzhou 730030,China)

机构地区：[1]西北民族大学生物医学研究中心甘肃省动物细胞技术创新中心,兰州730030 [2]西北民族大学生命科学与工程学院,兰州730030 [3]西北民族大学生物医学研究中心生物工程与技术国家民委重点实验室,兰州730030

出　　处：《黑龙江畜牧兽医》2020年第15期7-12,168,169,共8页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然科学基金项目(31760649,31660642);中央高校基本科研业务费专项(31920190020,31920190024)。

摘　　要：为了探究多不饱和脂肪酸对小尾寒羊前体脂肪细胞增殖的影响,试验采用Ⅰ型胶原酶消化法分离小尾寒羊尾部和皮下前体脂肪细胞进行原代和传代培养,并对其进行形态学观察,绘制生长曲线;诱导培养后,利用油红O染色法对其进行鉴定,观察其储脂特性;添加不同浓度(100,150,200,250,300,350μmol/L)α-亚麻酸(alpha-linolenic acid,ALA)和亚油酸(linoleic acid,LA),作用不同时间(1,3,5,7,9 d)后采用MTT法检测细胞增殖情况。结果表明:小尾寒羊尾部和皮下前体脂肪细胞均呈长梭形或不规则三角形,生长曲线均呈"S"形,尾部前体脂肪细胞倍增时间为22.04 h,显著短于皮下前体脂肪细胞(29.56 h);尾部前体脂肪细胞最大增殖密度为5.89×10^5个/mL,显著大于皮下前体脂肪细胞(2.97×10^5个/mL);不同部位前体脂肪细胞比生长速率无显著差异(P>0.05)。经诱导分化5 d后,细胞内均有可视脂滴,油红O染色呈红色。添加浓度为150μmol/L ALA或LA作用于前体脂肪细胞1 d,均能极显著促进细胞增殖;作用时间延长至3~9 d时,添加浓度为100~350μmol/L的ALA均能极显著抑制细胞增殖,而添加浓度为100μmol/L和150μmol/L的LA均能极显著促进细胞增殖,添加浓度为200~350μmol/L的LA均能极显著抑制细胞增殖。说明试验成功分离培养了小尾寒羊前体脂肪细胞,添加相同浓度ALA或LA作用于前体脂肪细胞1 d时,低浓度(100μmol/L和150μmol/L)均促进细胞增殖;作用3~9 d时,高浓度(200~350μmol/L)均抑制细胞增殖,且ALA的抑制作用较LA明显。In order to evaluate the effects of polyunsaturated fatty acids on the proliferation of precursor fat cells of small-tailed Han sheep, type I collagenase digestion method was used to isolate the tail and subcutaneous preadipocytes of the small-tailed Han sheep for primary and subculture, then the morphology of preadipocytes was observed and the growth curve was plotted. After induction culture, the preadipocytes were identified by oil red o staining method and their fat storage characteristics were observed. Then, the proliferation of cells treated with different concentrations(100, 150, 200, 250, 300, 350 μmol/L) of α-linolenic acid(alpha-linolenic acid, ALA) and linoleic acid(LA) was measured by MTT method at 1, 3, 5, 7 and 9 d. The results showed that the tail and subcutaneous preadipocytes of the small-tailed Han sheep were long fusiform or irregular triangular, and the growth curve was "S" shape. The doubling time of the tail preadipocyte was 22.04 h, which was significantly shorter than the subcutaneous preadipocyte(29.56 h). The maximum proliferation density of tail preadipocyte was 5.89×10^5 cells/mL, which was significantly larger than that of subcutaneous preadipocyte(2.97×10^5 cells/mL). There was no significant difference in the specific growth rate of preadipocyte in different parts(P>0.05). After 5 d of differentiation, there were visible lipid droplets in the cells, which were stained red by oil red o. The proliferation of preadipocytes treated with 150 μmol/L ALA or LA was significantly enhanced at 1 d. The proliferation of preadipocytes treated with 100-350 μmol/L ALA or 200-350 μmol/L LA was significantly inhibited during 3-9 d, and the proliferation of preadipocytes treated with 100 μmol/L and 150 μmol/L LA was significantly increased. These results indicated that the preadipocyte of Small Tail Han sheep were successfully isolated and cultured, low concentration(100 μmol/L and 150 μmol/L) of ALA or LA could promote the proliferation of preadipocytes at 1 d, and high concentrations of

关 键 词：小尾寒羊 前体脂肪细胞 生长曲线 油红O染色 α-亚麻酸(ALA) 亚油酸(LA) 

分 类 号：S826.89[农业科学—畜牧学] Q813.11[农业科学—畜牧兽医]