摘要
通过胶原酶、胰蛋白酶双消化法获取蛋鸡松果体细胞,分为对照(培养基)、酒精(溶剂对照)和褪黑素(MT)三组进行原代细胞培养,采用相对定量RT-PCR方法检测褪黑素(MT)对培养的松果体细胞促性腺激素释放激素-Ⅰ(GnRH-Ⅰ)基因表达的影响。结果表明:MTT检测正常培养的松果体细胞增殖速度第9d达高峰,11d缓降。与对照组比较,MT处理1周显著上调其表达丰度(P<0.05);与酒精溶剂组相比,MT处理1周具有上调其表达的趋势(P<0.1);对照组与酒精组之间没有明显的差异(P>0.05)。从而提示:体外培养松果体细胞的GnRH表达受MT的调控。
Abstract： Layer pinealocytes were isolated by using double digestion methods of collagenase and trypsin, and cells were divided into 3 groups, which were the control （medium）, alcohol（ solvent control）, and MT （mixed with alcohol） group. In order to observe the effects of Melatonin（MT） on the expression of GnRH - I gene in layers pinealocytes cultured in vitro, total RNA was extracted from cells on 11 th day of culture, and relative RT -PCR method was employed to detect GnRH -I mRNA expression using β -actin as the internal control. The results showed that the MTT value of pinealocytes increased progressively from 1 to 9 days of culture in vitro, andthere was a bit but not significant decrease on the 11 th day of culture compared to that of the 9th day. The expression abundance of GnRH - I mRNA in MT group were significantly higher than that of the control group （ P 〈 0.05 ）, and still has up - regulated the expression trend by MT treatment for seven days compared with alcohol group（P 〈0.1 ）, while there was no significant difference between control and alcohol groups （P 〉 0.05 ）. These results indicated that GnRH - I gene expression in layer pinealocytes cultured in vitro was significantly up regulated by exogenous MT treatment.
出处
《安徽科技学院学报》
2011年第6期1-5,共5页
Journal of Anhui Science and Technology University
基金
安徽科技学院重点学科建设项目(AKXK20101-2)
