摘要
该研究旨在探讨不同浓度的氢化可的松处理对奶牛乳腺上皮细胞(bovine mammary gland epithelial cells,BMECs)增殖、甘油三酯(triglyceride,TAG)合成、脂滴的形成以及乳脂肪合成相关基因表达的影响。采用单因子随机实验设计,以不添加氢化可的松组为对照组,处理组的氢化可的松浓度分别为1、10、100、1 000 ng/m L。各组细胞处理24 h后,通过四甲基偶氮唑盐(thiazoly blue tetrazolium,MTT)比色法检测细胞活力;利用试剂盒检测胞内TAG的含量;使用油红O染色法检测细胞内乳脂球的合成;采用实时定量PCR法检测乳脂合成相关基因的表达。结果表明,氢化可的松处理对BMECs增殖无显著影响(P>0.05);与对照组相比,100 ng/m L氢化可的松组能够显著提高TAG的合成量、增加胞内脂滴的形成以及过氧化物酶体增殖物激活受体γ(peroxisome proliferator activated receptorγ,PPARγ)的基因表达(P<0.05);10 ng/m L和100 ng/m L氢化可的松组能够显著上调乙酰辅酶A羧化酶(acetyl-Co A carboxylase,ACC)、二酰甘油酰基转移酶(diacylgycerol acyltransferase,DGAT)的基因表达(P<0.05);1、10、100 ng/m L氢化可的松组能够显著上调脂肪酸合酶(fatty acid synthase,FASN)基因表达(P<0.05),且1 ng/m L的促进效果最明显,而1 000 ng/m L则显著抑制了FASN的表达(P<0.05)。该研究结果说明,氢化可的松能够促进BMECs乳脂肪合成,且100 ng/m L的剂量对乳脂合成的促进效果最佳。
The aim of this study was to explore the effects of hydrocortisone（HYD） on proliferation, synthesis of triglyceride, formation of lipid droplets and expression of genes involved in milk fat synthesis of bovine mammary gland epithelial cells（BMECs）. A single factor randomized trial design was applied. Different concentrations（0, 1, 10, 100, 1 000 ng/m L） of HYD were added in culture medium of BMECs for 24 h. Cell viability was detected by thiazoly blue tetrazolium（MTT）. Triglyceride（TAG） content was measured by TAG determination kit. Expression of genes involved in milk fat synthesis were measured by real time quantitative PCR（RT-q PCR）. The results showed that the activity of cell proliferation was not influenced by supplementation of HYD（P〉0.05）. Compared with the control group, 100 ng/m L HYD significantly increased the concentration of TAG and the formation of intracellular fat droplets, the expression of peroxisome proliferator activated receptor γ（PPARγ） was also upregulated（P〈0.05）. 10 ng/m L and 100 ng/m L HYD significantly increase the gene expressions of acetyl-Co A carboxylase（ACC） and diacylgycerol acyltransferase（DGAT）（P〈0.05）; 1, 10, 100 ng/m L HYD significantly upregulated the m RNA level of fatty acid synthase（FASN）, whereas 1 000 ng/m L HYD was significantly downregulated the m RNA level of FASN（P〈0.05）. In conclusion, HYD treatment can promote milk fat synthesis in BMECs and the 100 ng/m L HYD is an optimal level considering its improvement effects on milk fat synthesis of BMECs.
作者
孙梅
李大彪
邢媛媛
张花
王卫云
李子健
Sun Mei Li Dabiao Xing Yuanyuan Zhang Hua Wang Weiyun Li Zijian(College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China)
出处
《中国细胞生物学学报》
CAS
CSCD
2017年第9期1156-1164,共9页
Chinese Journal of Cell Biology
基金
国家自然科学基金(批准号:31360559)资助的课题~~
关键词
氢化可的松
奶牛乳腺上皮细胞
乳脂合成
hydrocortisone
bovine mammary epithelial cells
milk fat synthesis
