白细胞介素8在妊娠早期小鼠子宫内膜的表达及雌、孕激素的调节作用
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摘要
目的:研究白细胞介素8(IL-8)、雌激素受体(ER)和孕激素受体(PR)在妊娠早期小鼠子宫内膜的表达,揭示雌、孕激素对着床前小鼠子宫内膜IL-8蛋白表达的调节作用。方法:应用半定量逆转录-聚合酶链式反应(RT-PCR)技术,检测妊娠早期(妊娠1d、4d、5d、6d)小鼠子宫内膜IL-8mRNA的表达;利用免疫组织化学染色及图像分析技术对妊娠1d、4d、5d、6d小鼠子宫内膜ER和PR的表达进行定位及半定量分析;利用小鼠胚泡延迟着床模型,研究胚泡着床前雌、孕激素对小鼠子宫内膜IL-8表达的调节作用。结果:RT-PCR结果显示:妊娠1d、4d、5d、6d小鼠子宫内膜均有IL-8mRNA表达,妊娠1d表达最弱,妊娠4d表达最强,妊娠5d表达降低,妊娠6d表达继续下降。免疫组织化学染色结果显示:ER、PR蛋白主要表达于小鼠子宫内膜腔上皮细胞的核膜、核仁及胞质中,ER于妊娠1d表达最强,妊娠4d最弱,妊娠5d、6d表达又逐渐增强;PR于妊娠1d表达较弱,妊娠4d表达最弱,妊娠5d表达增强,妊娠6d表达明显增强。小鼠胚泡延迟着床实验显示:切除双侧卵巢后,P_4组(单独予以孕酮)IL-8表达低于E_2+P_4组(联合应用雌二醇和孕酮)及对照组; E_2+P_4组IL-8表达上调,但低于对照组。结论:1.IL-8mRNA在妊娠早期小鼠子宫内膜持续表达,于妊娠4d表达最强;2.ER、PR蛋白在妊娠早期小鼠子宫内膜腔上皮持续表达;3.雌、孕激素通过其受体发挥对母胎界面IL-8表达的调节作用;4.卵巢切除后,联合应用雌、孕激素能上调着床前小鼠子宫内膜IL-8的蛋白表达。
Objective:To study the expressions of interleukin-8(IL-8),estrogen receptor(ER)and progestogen receptor(PR)in mouse endometrium during early pregnancy,to reveal the regulation of estrogen and progestogen on IL-8 protein of mouse endometrium in preimplantation. Methods:The expression of IL-8mRNA in mouse endometrium of early pregnancy was measured by semi-quantitative reverse transcriptional-polymerase chain reaction(RT-PCR).ER and PR expressions in mouse endometrium on Day 1,Day 4,Day 5 and Day 6 were shown and measured by immunohistochemical method and image analysis.Using delayed implantation model to study the regulation of estrogen and progestogen on IL-8 expression before blastocyst implantation. Results:IL-8mRNA continually expressed in mouse endometrium during early pregnancy,the expression of IL-8mRNA was the weakest on Day 1,the strongest on Day 4,then reduced on Day 5,and kept decreasing on Day 6.Immunohistochemical stain discovered that ER and PR proteins were mainly located in the nuclear membrane,nucleolus and cytoplasm in pregnant mouse endometrium.The expression of ER was the strongest on Day 1,the weakest on Day 4,and gradually increased on Day 5 and Day 6.The expression of PR was weak on Day 1,the weakest on Day 4,enhanced on Day 5,and obviously strengthened on Day 6.In delayed implantation model,the expression of IL-8 was significantly decreased in P_4 group(only treated by progesterone after ovariectomy),and lower than that in E_2 + P_4(treated by estradiol and progesterone after ovariectomy)and control group.The level of IL-8 was up-regulated by treated with E_2 and P_4,but lower than control . Conclusions : 1. IL-8mRNA continuously expressed in mouse endometrium during early pregnancy,and strongest on Day 4. 2. ER and PR proteins were persistently expressed in the luminal epithelium of mouse endometrium during early pregnancy.3. Estrogen and progestogen modulated the expression of IL-8 by their receptors at the maternal-fetal interface.4. Applying progesterone associated with estrogen could up-regulate the expression of IL-8 protein in mouse endometrium of preimpalntation after ovariectomy.
Objective:To study the expressions of interleukin-8(IL-8),estrogen receptor(ER)and progestogen receptor(PR)in mouse endometrium during early pregnancy,to reveal the regulation of estrogen and progestogen on IL-8 protein of mouse endometrium in preimplantation. Methods:The expression of IL-8mRNA in mouse endometrium of early pregnancy was measured by semi-quantitative reverse transcriptional-polymerase chain reaction(RT-PCR).ER and PR expressions in mouse endometrium on Day 1,Day 4,Day 5 and Day 6 were shown and measured by immunohistochemical method and image analysis.Using delayed implantation model to study the regulation of estrogen and progestogen on IL-8 expression before blastocyst implantation. Results:IL-8mRNA continually expressed in mouse endometrium during early pregnancy,the expression of IL-8mRNA was the weakest on Day 1,the strongest on Day 4,then reduced on Day 5,and kept decreasing on Day 6.Immunohistochemical stain discovered that ER and PR proteins were mainly located in the nuclear membrane,nucleolus and cytoplasm in pregnant mouse endometrium.The expression of ER was the strongest on Day 1,the weakest on Day 4,and gradually increased on Day 5 and Day 6.The expression of PR was weak on Day 1,the weakest on Day 4,enhanced on Day 5,and obviously strengthened on Day 6.In delayed implantation model,the expression of IL-8 was significantly decreased in P_4 group(only treated by progesterone after ovariectomy),and lower than that in E_2 + P_4(treated by estradiol and progesterone after ovariectomy)and control group.The level of IL-8 was up-regulated by treated with E_2 and P_4,but lower than control . Conclusions : 1. IL-8mRNA continuously expressed in mouse endometrium during early pregnancy,and strongest on Day 4. 2. ER and PR proteins were persistently expressed in the luminal epithelium of mouse endometrium during early pregnancy.3. Estrogen and progestogen modulated the expression of IL-8 by their receptors at the maternal-fetal interface.4. Applying progesterone associated with estrogen could up-regulate the expression of IL-8 protein in mouse endometrium of preimpalntation after ovariectomy.
引文
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