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来自对蜕膜细胞反应敏感性不同的子宫的培养大鼠子宫内膜基质细胞分泌纤溶酶原激活物。

Secretion of plasminogen activator by cultured rat endometrial stromal cells from uteri differentially sensitized for the decidual cell reaction.

作者信息

Kennedy T G, Ross H E, Barbe G J, Shu M A, Zhang X

机构信息

Department of Physiology, University of Western Ontario, London, Canada.

出版信息

Mol Reprod Dev. 1998 Mar;49(3):268-76. doi: 10.1002/(SICI)1098-2795(199803)49:3<268::AID-MRD7>3.0.CO;2-M.

Abstract

Endometrial stromal cells from rat uteri differentially sensitized for the decidual cell reaction in vivo and which undergo differing degrees of decidualization in vitro were cultured and plasminogen activator (PA) in the medium determined. The cells were obtained by enzymatic dispersion from the uteri of ovariectomized, steroid-treated rats at the equivalent of day 4, 5, or 6 of pseudopregnancy or on day 5 from rats treated on day 4 with 0, 0.3, or 1.0 microgram estradiol (low, intermediate, or high dose of estradiol, respectively) and cultured for 24, 48, or 72 hr. For cells from day 4, 5, and 6 uteri cultured under control conditions, PA activity in the medium was greatest for day 5 cells, which were from uteri maximally sensitized for decidualization both in vivo and in vitro. By contrast, for cells from low-, intermediate-, and high-estradiol uteri, PA activity in the medium was greatest for the high-estradiol cells; these cells do not undergo decidualization in vivo or in vitro to the same extent as intermediate-estradiol cells. Indomethacin, an inhibitor of prostaglandin (PG) synthesis, reduced PGE2 accumulation to nondetectable amounts and for most cultures decreased PA activity in the medium, suggesting that endogenous PG production regulated in part PA secretion under control conditions. The addition of PGE2 with indomethacin increased PA activities above those under control conditions, but activities were still lower for day 4 and 6 cells compared with day 5 cells, and for low- and intermediate-estradiol cells compared with high-estradiol cells. This indicates that the differences in PA secretion are not explainable by differences in PGE2 production. Northern blot analysis of RNA from cells cultured for 72 hr under control conditions did not reveal significant differences in steady-state concentrations of mRNA for urokinase-type PA or plasminogen activator inhibitor 1, but those for tissue-type PA were lower in day 6 cells compared with day 4 and 5 cells. It is concluded that PA activity secreted by the cultured endometrial stromal cells, although controlled in part by the endocrine milieu to which they were exposed prior to culture, does not simulate decidualization in vitro and, therefore, that PA activity is not a marker for decidualization in vitro.

摘要

培养来自大鼠子宫的子宫内膜基质细胞,这些细胞对体内蜕膜细胞反应的敏感性不同,且在体外经历不同程度的蜕膜化,并测定培养基中的纤溶酶原激活物(PA)。细胞通过酶分散法从卵巢切除、经类固醇处理的大鼠子宫中获得,这些大鼠处于假孕第4、5或6天的等效阶段,或者在第4天分别用0、0.3或1.0微克雌二醇(分别为低、中、高剂量雌二醇)处理的大鼠在第5天的子宫中获取,并培养24、48或72小时。对于在对照条件下培养的来自第4、5和6天子宫的细胞,培养基中的PA活性在第5天的细胞中最高,这些细胞来自在体内和体外对蜕膜化均高度敏感的子宫。相比之下,对于来自低、中、高雌二醇子宫的细胞,培养基中的PA活性在高雌二醇细胞中最高;这些细胞在体内或体外的蜕膜化程度不如中雌二醇细胞。吲哚美辛是一种前列腺素(PG)合成抑制剂,它将PGE2积累量降低到无法检测的水平,并且在大多数培养物中降低了培养基中的PA活性,这表明在对照条件下内源性PG的产生部分调节了PA的分泌。与吲哚美辛一起添加PGE2可使PA活性高于对照条件下的活性,但与第5天的细胞相比,第4天和第6天细胞的活性仍然较低,并且与高雌二醇细胞相比,低、中雌二醇细胞的活性也较低。这表明PA分泌的差异不能用PGE2产生的差异来解释。对在对照条件下培养72小时的细胞的RNA进行Northern印迹分析,未发现尿激酶型PA或纤溶酶原激活物抑制剂1的mRNA稳态浓度有显著差异,但与第4天和第5天的细胞相比,第6天细胞中组织型PA的mRNA稳态浓度较低。得出的结论是,培养的子宫内膜基质细胞分泌的PA活性虽然部分受其培养前暴露的内分泌环境控制,但在体外并不模拟蜕膜化,因此,PA活性不是体外蜕膜化的标志物。

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