本文選題：糖皮質(zhì)激素 + 11βHSD-1��； 參考：《山東農(nóng)業(yè)大學》2015年碩士論文

【摘要】：宮頸成熟(Cervical ripening)是正常分娩的必要條件,不是繼發(fā)于子宮收縮的被動過程,而是一個多種內(nèi)分泌因素參與的主動耗能、炎性細胞浸潤的過程。各種激素和炎癥介質(zhì)通過直接或間接調(diào)節(jié)宮頸內(nèi)組織學、生物化學、及形態(tài)學變化來發(fā)揮它們特有的生理功能使宮頸成熟軟化。其中糖皮質(zhì)激素及其受體的作用一直備受關注,但其調(diào)控機制尚不清楚。本研究旨在對這一機制進行初步闡釋。糖皮質(zhì)激素(Glucocorticoid,GC)是腎上腺皮質(zhì)層束狀帶合成和分泌的一類甾體激素,在機體的代謝和應激等過程中起著重要的作用。糖皮質(zhì)激素通過與其受體結合發(fā)揮生物學效應,其在組織的表達量在一定程度上決定了組織對GC的敏感性。11βHSD-1和11βHSD-2是調(diào)節(jié)糖皮質(zhì)激素活性的轉化酶,二者作用相反,分別可以活化/失活GC,共同調(diào)節(jié)局部組織細胞內(nèi)糖皮質(zhì)激素的活性與有效濃度,所以,局部組織對糖皮質(zhì)激素的的敏感性還取決于糖皮質(zhì)激素代謝酶的表達量與反應方向。糖皮質(zhì)激素是參與分娩的重要激素,通過與其受體結合發(fā)揮生物學效應,但其對子宮頸成熟擴張的調(diào)控機制尚不清楚。本研究旨在揭示糖皮質(zhì)激素(GC)及糖皮質(zhì)激素受體(GR)、11β羥基類固醇脫氫酶1和2(11βHSD-1和2)對圍產(chǎn)期小鼠子宮頸成熟的影響和機制。實驗選取圍產(chǎn)期5個時間點,分別為產(chǎn)前48 h(-48 h)、產(chǎn)前24 h(-24 h)、分娩時(0 h)、產(chǎn)后12 h(+12 h)和產(chǎn)后48 h(+48 h),通過ELISA檢測小鼠血清皮質(zhì)酮濃度變化;用Western Blot法檢測兔抗人11βHSD-1和-2多克隆抗體及兔抗人糖皮質(zhì)激素受體(NR3C1)多克隆抗體在圍產(chǎn)期小鼠子宮頸組織中的特異性;用免疫組織化學法分別對NR3C1、11βHSD-1和11βHSD-2進行進行免疫定位和半定量檢測。結果顯示:ELISA結果顯示圍產(chǎn)期糖皮質(zhì)激素的濃度呈先升高后下降趨勢,分娩時達到峰值(P?0.01);Western Blot法檢測出在35KDa、44KDa和86KDa處分別有11βHSD-1和-2以及NR3C1蛋白特異性條帶的存在。11βHSD-1、11βHSD-2和GR的免疫組化陽性染色主要分布于子宮頸的腔上皮、基質(zhì)、血管和平滑肌細胞,且11βHSD-1/11βHSD-2平均光密度的比值呈現(xiàn)先升高再下降趨勢,分娩時比值最高;GR的表達量也是分娩時最高,分娩前后較低的趨勢(P?0.01)。以上結果提示分娩時11βHSD-1合成的糖皮質(zhì)激素及其受體大量增加,參與子宮頸的成熟軟化,分娩后外周血液循環(huán)和子宮頸局部組織糖皮質(zhì)激素水平降低且糖皮質(zhì)激素受體表達量下降,子宮頸恢復到產(chǎn)前的狀態(tài)。因此,11βHSD-1合成的糖皮質(zhì)激素通過其受體在子宮頸的成熟過程中起著關鍵性作用。
[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.
【學位授予單位】：山東農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S857.2

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