摘要
为了研究17β-雌二醇对Ghrelin表达量的影响,根据绵羊Ghrelin基因mRNA的保守序列设计特异性引物,通过体外培养绵羊输卵管上皮细胞,提取经17β-雌二醇处理的绵羊输卵管上皮细胞总RNA,并反转录成cDNA,以β-actin基因为内参,采用SYBR GreenⅠ染料法进行实时荧光定量PCR检测。结果显示,绵羊输卵管上皮细胞经17β-雌二醇处理后,Ghrelin基因的表达量有显著性增加,在处理后第6小时表达量达到最高。结论,17β-雌二醇对绵羊输卵管上皮细胞表达Ghrelin有显著的促进作用,这为后续试验研究打下了良好基础。
In order to study the effects of 17β-estradiol on the expression of ghrelin gene,the spec-ific primers were designed according to the conserved sequences of ghrelin gene.Total RNA was extracted from ovine oviduct epithelial cells stimulated by 17β-estradiol,then it was reversely transcribed into c DNA.The pair of primer was designed for developing a SYBR Green I quantitative real-time PCR for detection of ghrelin cytokines of ovine,while the β-actin gene was used as an internal control.In result,because ovine oviduct epithelial cells were stimulated by 17β-estradiol,the expression of ghrelin gene was significantly increased and the expression of the gene was the highest at hour 6post-treatment.In conclusion,17β-estradiol significantly promotes the expression of ghrelin gene in the ovine oviduct epithelial cells and the above-mentioned results lay a good foundation for subsequent experimental studyies.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2017年第8期1056-1061,共6页
Chinese Veterinary Science
基金
国家自然科学基金项目(30660128
30860201)
