【作者】 武岳；

【导师】 曹贵方； 曹金山；

【作者基本信息】 内蒙古农业大学 ， 基础兽医学， 2010， 硕士

【摘要】 环氧合酶(cyclooxygenase ,COX)是动物组织合成前列腺素(prostaglandin, PG)的限速酶,目前已知它存在两个同功酶(COX-1和COX-2)。许多对前列腺素的研究已经证实,前列腺素是哺乳动物体内作用极为广泛的局部激素,影响生殖过程的多个环节,在哺乳动物的排卵、受精、分娩、着床等生殖过程中发挥重要的作用,其中也涉及到对输卵管功能的调节。但是对于前列腺素(prostaglandin, PG)合成的研究主要还是集中在环氧合酶(cyclooxygenase ,COX)上,目前尚未有关于雌性动物输卵管上皮细胞的环氧合酶表达和雌激素、孕激素关系的研究报道。为阐明雌激素对输卵管上皮细胞中环氧合酶的表达调控机理,在本文应用实时荧光定量RT-PCR技术(real-time RT-PCR)和蛋白印迹技术(Western blot),采用体外培养奶牛输卵管上皮细胞(bovine oviduct epithelial cells,BOECs)并且建立传代细胞系的方法,运用体外方法研究了雌激素和孕激素对奶牛输卵管上皮细胞环氧合酶表达的影响。在体外培养的BOECs中,分不同浓度、不同时间添加17β-雌二醇、孕酮,研究BOECs中环氧合酶的表达量和雌激素、孕激素的关系。实验结果表明,添加不同浓度的17-β-雌二醇培养24 h,COX-1mRNA在100pg/ml(10-10g/mL)时表达量最高, COX-2 mRNA在100 pg/mL(10-10g/ml)时表达量达到最高值。添加不同浓度的孕酮诱导对体外培养的BOECs中COX-1、COX-2 mRNA转录,COX-1在1ng/ml(10-9 g/ml)时高表达, COX2 mRNA在100pg/ml(10-10g/ml)时高表达。结果与在奶牛的体内正常的雌、孕激素生理浓度相关。而添加10pg/mL的17-β-雌二醇、10ng/mL孕酮诱导诱导培养不同时间,对BOECs中COX-1、COX-2mRNA转录有影响,2h时表达量均较高。上述结果提示, COX-1和COX-2的表达受到雌、孕激素浓度的调节,而且17-β-雌二醇影响比孕酮明显, COX-2受的影响要比COX-1大,与COX-2是诱导型酶受诱导因子的影响较大的原因有关。更多还原

【Abstract】 Cyclooxygenase (COX) is a rate-limiting enzyme for animal tissue produced prostaglandin. It has two currently known isoenzymes COX-1 and COX-2. Many research results show that prostaglandin is a local hormone performing a wide range of functions in mammals. It plays an important role in many stages of the birth process such as ovulation, fecundation, labor and rooting, etc., including the regulation of fallopian tube functions. Studies into the adjusting function of PG have mainly focused on COXs, and there has been no report about research on the interrelations between the COX expression of female oviduct epithelial cells, and estrogen and progesterone. In order to clarify how estrogen regulates COX expression of female animal oviduct epithelial cells, this research adopts the real-time RT-PCR and Western blot methods. BOECs are cultivated extra-corporally and method to establish the regenerating cells is found. Extra-corporal method is used to discover the impact of estrogen and progesterone on the COX expression of female animal oviduct epithelial cells. BOECs cultured in vitro are of different concentrations.17β-estradiol and progesterone are treated at different times. The purpose is to find out the interrelation between the expressed amount of COX and estrogen as well as progesterone. Study results show adding 17β-estradiol of different concentrations, and after 24 hours of culturing, COX-1mRNA reaches the highest point in 100pg/ml(10-10g/ml). COX-2 mRNA reaches the highest point in 1000 pg/ml(10-9g/ml). Copying COX-1, COX-2 and mRNA in BOECs is affected by the time when 17β-estradiol is added. However, there is no obvious regularity regarding concentration of liquid. Adding progesterone of different concentrations can help copy COX-1, COX-2 and mRNA in BOECs. COX-1runs high in 1ng/ml(10-9 g/ml)while COX-2 mRNA in 10ng/ml. Statistics acquired from adding 10ng/mL progesterone at different times demonstrate less regularity than concentration. Study results show the expression of COX-1 and COX-2 are regulated by the concentration of estrogen and progesterone. And COX-1 is more seriously affected than COX-2. 17β-estradiol makes a greater change than progesterone. This is because COX-2 is more sensitive to inducing agents. As the amount of progesterone in the subject animal is regulated by estradiol, much of its power depends on estradiol.更多还原