围产期小鼠子宫颈中糖皮质激素受体、11β羟基类固醇脱氢酶1和2的免疫定位

发布时间：2018-05-30 22:05

本文选题：糖皮质激素 + 11βHSD-1　；参考：《山东农业大学》2015年硕士论文

【摘要】：宫颈成熟(Cervical ripening)是正常分娩的必要条件,不是继发于子宫收缩的被动过程,而是一个多种内分泌因素参与的主动耗能、炎性细胞浸润的过程。各种激素和炎症介质通过直接或间接调节宫颈内组织学、生物化学、及形态学变化来发挥它们特有的生理功能使宫颈成熟软化。其中糖皮质激素及其受体的作用一直备受关注,但其调控机制尚不清楚。本研究旨在对这一机制进行初步阐释。糖皮质激素(Glucocorticoid,GC)是肾上腺皮质层束状带合成和分泌的一类甾体激素,在机体的代谢和应激等过程中起着重要的作用。糖皮质激素通过与其受体结合发挥生物学效应,其在组织的表达量在一定程度上决定了组织对GC的敏感性。11βHSD-1和11βHSD-2是调节糖皮质激素活性的转化酶,二者作用相反,分别可以活化/失活GC,共同调节局部组织细胞内糖皮质激素的活性与有效浓度,所以,局部组织对糖皮质激素的的敏感性还取决于糖皮质激素代谢酶的表达量与反应方向。糖皮质激素是参与分娩的重要激素,通过与其受体结合发挥生物学效应,但其对子宫颈成熟扩张的调控机制尚不清楚。本研究旨在揭示糖皮质激素(GC)及糖皮质激素受体(GR)、11β羟基类固醇脱氢酶1和2(11βHSD-1和2)对围产期小鼠子宫颈成熟的影响和机制。实验选取围产期5个时间点,分别为产前48 h(-48 h)、产前24 h(-24 h)、分娩时(0 h)、产后12 h(+12 h)和产后48 h(+48 h),通过ELISA检测小鼠血清皮质酮浓度变化;用Western Blot法检测兔抗人11βHSD-1和-2多克隆抗体及兔抗人糖皮质激素受体(NR3C1)多克隆抗体在围产期小鼠子宫颈组织中的特异性;用免疫组织化学法分别对NR3C1、11βHSD-1和11βHSD-2进行进行免疫定位和半定量检测。结果显示:ELISA结果显示围产期糖皮质激素的浓度呈先升高后下降趋势,分娩时达到峰值(P?0.01);Western Blot法检测出在35KDa、44KDa和86KDa处分别有11βHSD-1和-2以及NR3C1蛋白特异性条带的存在。11βHSD-1、11βHSD-2和GR的免疫组化阳性染色主要分布于子宫颈的腔上皮、基质、血管和平滑肌细胞,且11βHSD-1/11βHSD-2平均光密度的比值呈现先升高再下降趋势,分娩时比值最高;GR的表达量也是分娩时最高,分娩前后较低的趋势(P?0.01)。以上结果提示分娩时11βHSD-1合成的糖皮质激素及其受体大量增加,参与子宫颈的成熟软化,分娩后外周血液循环和子宫颈局部组织糖皮质激素水平降低且糖皮质激素受体表达量下降,子宫颈恢复到产前的状态。因此,11βHSD-1合成的糖皮质激素通过其受体在子宫颈的成熟过程中起着关键性作用。
[Abstract]:Cervical ripeningis a necessary condition for normal delivery. It is not a passive process secondary to uterine contraction, but a process of active energy dissipation and inflammatory cell infiltration involving a variety of endocrine factors. Hormones and inflammatory mediators play their unique physiological functions by directly or indirectly regulating cervical histology, biochemistry, and morphology to make the cervix mature and soften. The role of glucocorticoid and its receptors has been concerned, but its regulatory mechanism is not clear. The purpose of this study is to explain this mechanism. Glucocorticoid (GCC) is a steroid hormone synthesized and secreted by the adrenal cortical bundles, which plays an important role in the metabolism and stress of the body. Glucocorticoids play a biological role by binding to their receptors. The expression of glucocorticoids in the tissues determines to a certain extent that the sensitivity of tissues to GC. 11 尾 HSD-1 and 11 尾 HSD-2 are invertase regulating glucocorticoid activity. The activity and effective concentration of glucocorticoid were regulated by activating / inactivating GCrespectively. Therefore, the sensitivity of local tissue to glucocorticoid was also determined by the expression and reaction direction of glucocorticoid metabolic enzyme. Glucocorticoid is an important hormone involved in labor. It plays a biological role by binding to its receptor, but its regulatory mechanism on cervical maturation and dilatation is still unclear. The purpose of this study was to investigate the effect and mechanism of glucocorticoid (GCC) and glucocorticoid receptor GRN 11 尾 hydroxy steroid dehydrogenase 1, 2G 11 尾 HSD-1 and 2) on cervix maturation in perinatal mice. Five perinatal time points were selected: 48 h prepartum, 48 h prepartum, 24 h prepartum, 0 h postpartum, 12 h postpartum (12 h) and 48 h postpartum (48 h). The changes of serum corticosterone concentration in mice were detected by ELISA. The specificity of rabbit anti-human 11 尾 HSD-1 and 2 polyclonal antibody and rabbit anti-human glucocorticoid receptor NR3C1 polyclonal antibody in perinatal mouse cervix was detected by Western Blot method. Immunolocalization and semi-quantitative detection of NR3C1O11 尾 HSD-1 and 11 尾 HSD-2 were performed by immunohistochemical method. The results showed that the concentration of glucocorticoid in perinatal period increased first and then decreased. The presence of 11 尾 HSD-1 and 2 and NR3C1 protein specific bands at 35K Dawei 44KDa and 86KDa were detected by Western Blot. The immunohistochemical staining of 11 尾 HSD-1 and gr were mainly distributed in cervical cavity epithelium, stroma, blood vessels and smooth muscle cells. The ratio of average optical density of 11 尾 HSD-1/11 尾 HSD-2 increased at first and then decreased, the highest expression of gr was also the highest in parturition, and the tendency was lower before and after parturition. The above results suggest that the glucocorticoid and its receptors synthesized by 11 尾 HSD-1 increase significantly during labor, and participate in the maturation and softening of cervix. The levels of glucocorticoid and the expression of glucocorticoid receptor decreased after delivery, and the cervix returned to antepartum state. Therefore, glucocorticoid synthesized by 11 尾 HSD-1 plays a key role in cervix maturation through its receptors.
【学位授予单位】：山东农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S857.2

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