畜牧兽医学报 ›› 2014, Vol. 45 ›› Issue (10): 1663-1670.doi: 10.11843/j.issn.0366-6964.2014.10.012

• 动物营养 • 上一篇    下一篇

氢化可的松对奶牛乳腺上皮细胞酪蛋白合成的影响

田青1,2,王洪荣2*,王梦芝2   

  1. (1.江苏食品药品职业技术学院,淮安 223003; 2.扬州大学动物科学与技术学院,扬州 225009)
  • 收稿日期:2014-01-22 出版日期:2014-10-23 发布日期:2014-10-23
  • 通讯作者: 王洪荣,教授,博士生导师,E-mail: hrwang@yzu.edu.cn
  • 作者简介:田青(1980-) ,女,江苏淮安人,博士生,从事反刍动物营养研究,E-mail: tianqing800312@163.com
  • 基金资助:

    国家重点基础研究发展规划(973 计划) 项目( 2011CB100803);江苏高校优势学科建设工程资助项目;江苏省研究生创新工程项目(CXZZ11_0989)

Effects of HYD on the Synthesis of Casein in Mammary Epithelial Cells of Holstein Cows in vitro

TIAN Qing1,2,WANG Hong-rong2*,WANG Meng-zhi2   

  1. (1.Jiangsu Food & Pharmaceutical Science College,Huaian 223003,China;2.College of Animal Science and Technology,Yangzhou University,Yangzhou 225009,China)
  • Received:2014-01-22 Online:2014-10-23 Published:2014-10-23

摘要:

旨在研究培养基中添加不同浓度的氢化可的松对奶牛乳腺上皮细胞生长、κ-酪蛋白(CSN3) 和糖皮质激素受体(NR3C1) 基因表达及其κ-酪蛋白和总酪蛋白相对含量的影响。选用中国荷斯坦奶牛的乳腺上皮细胞进行体外培养,在以无血清无激素的生长培养基为对照的基础上,分别添加氢化可的松1、10、100 和1 000 ng•mL-1,测定CSN3和NR3C1基因表达量以及κ-酪蛋白和总酪蛋白相对含量的变化。结果表明: 1)培养时间<20 h,1~1 000 ng•mL-1氢化可的松对健康奶牛乳腺上皮细胞增殖的影响差异不显著(P>0.05),但1 000 ng•mL-1组细胞增殖较慢;培养时间>20 h,1 000 ng•mL-1的氢化可的松显著抑制了奶牛乳腺上皮细胞的增殖(P<0.05)。2)1~100 ng•mL-1的氢化可的松对健康奶牛乳腺上皮细胞CSN3、NR3C1基因表达具有显著地促进作用(P<0.05),其中1~10 ng•mL-1的氢化可的松能够显著增加奶牛乳腺上皮细胞κ-酪蛋白和总酪蛋白的相对含量(P<0.05)。3)氢化可的松的促进乳蛋白合成作用具有一定的剂量依赖关系,小剂量的氢化可的松(1 ng•mL-1)有利于奶牛乳腺上皮细胞乳蛋白的合成,随着添加剂量的增加,尤其是大于1 000 ng•mL-1,它的促乳蛋白合成作用逐渐减弱,甚至抑制了奶牛乳腺细胞的增长进而抑制乳腺乳蛋白的合成。结果提示,随着氢化可的松浓度的增加,CSN3 和NR3C1 基因的表达量及CSN3和总酪蛋白相对含量均呈下降的趋势,其中以1 ng•mL-1 的添加效果最好。

Abstract:

This study was conducted to investigate the effects of hydrocortisone (HYD) on growth,expressions of κ-casein (CSN3) and hydrocortisone receptor (NR3C1) genes and the relative quantities of κ-casein and total casein in bovine mammary epithelial cells.Bovine mammary epithelial cells from Chinese Holstein were used in an in vitro cultivation.Cells were cultured in a growth culture medium (without serum and hormone,control group),and the growth culture medium supplemented with 0,1,10,100 and 1 000 ng•mL-1 HYD,respectively.Changes of expression levels of CSN3 and NR3C1 genes and relative content of CSN3 and total casein were determined.The results showed as follows: 1) The cells proliferation in 1 000 ng•mL-1 group was lower than these in the others in numerical value before 20 h,but significant difference was found in cell proliferation between groups after 20 h (P<0.05).2) The expression of CSN3 and NR3C1 gene were increased in 1-100 ng•mL-1 HYD significantly (P<0.05).The relative quantities of κ-casein and total casein in 1 and 10 ng•mL-1 groups were significantly higher than these in the other groups (P<0.05).3) A certain dose-response relationship existed between HYD and casein.That meant,small dose of HYD (1 ng•mL-1) was more advantageous to promote the milk protein synthesis,but with the increase of HYD concentration,the promoting effects were gradually weakened and even reverse with the increase of HYD concentration (1 000 ng•mL-1).The results indicated that with the increase of HYD concentration,the expression levels of CSN3 and NR3C1 genes and the relative quantities of CSN3 and total casein tended to be decreased,the optimal level was 1 ng•mL-1.

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