ESTROGEN PRODUCTION BY BOVINE BINUCLEATE AND MONONUCLEATE TROPHOBLASTIC CELLS IN-VITRO
ESTROGEN PRODUCTION BY BOVINE BINUCLEATE AND MONONUCLEATE TROPHOBLASTIC CELLS IN-VITRO
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DOI：
10.1095/biolreprod51.3.486
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发表时间：
1994-09-01
影响因子：
3.6
通讯作者：
REIMERS, TJ
中科院分区：
文献类型：
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作者：
MATAMOROS, RA;CAAMANO, L;REIMERS, TJ
长期以来，牛胎盘一直被认为是类固醇激素的来源。我们进行了三个实验来比较牛单核和双核滋养层细胞产生雌激素的情况，并检测了皮质醇、孕酮、孕烯醇酮、睾酮和雄烯二酮的影响。在第一个实验中，通过单位重力沉降法从6个酶分散的胎盘中纯化了150-180天的双核滋养层细胞。细胞(8×10(5)/ml)首先在37℃下与单独的199培养液(M199/6h)或10(-7)M皮质醇(皮质醇/6h)孵育6h。用10(-7)M孕酮、10(-7)M孕烯醇酮、10(-7)M睾酮、10(-7)M睾酮或M199代替培养液，第二次孵育4h后，在M199和皮质醇中孵育6h的细胞雌二醇的产生没有差异，孕酮和孕烯醇酮也不影响雌二醇的产生。睾酮增加(p<；0.05)雌二醇的产生。在M199和皮质醇中培养6小时的细胞中雌酮的产生没有差别；雌酮的产生不受孕酮、孕烯醇酮或睾酮的影响。从妊娠165到180天的胎盘中提纯单核和双核细胞，并用于另外两个实验。在第一组中，先用199(M199)或10(-7)M皮质醇培养6h，培养的是浓缩的双核和单核细胞。然后用10(-7)M睾酮、10(-7)M雄烯二酮或M199代替培养液，继续孵育4小时。在第一次G-h孵育中，单核细胞比双核细胞产生更多的雌酮(p<；0.05)；皮质醇不影响这种产生。单核细胞产生雌酮不受睾酮或雄烯二酮的影响。培养前6h的细胞产生雌二醇在不同处理或不同细胞类型之间没有差异。睾酮增加了双核细胞产生雌二醇(P<；0.05)，但不能增加单核细胞产生雌二醇。在这一系列实验的第三组中，双核和单核细胞与M199、孕酮或孕烯醇酮孵育连续10小时。孕酮和孕烯醇酮不影响单核细胞产生雌酮。然而，单核细胞的雌酮产量高于双核细胞(p<；0.05)。雌二醇的产生在不同处理或不同细胞类型之间没有差异。这些数据表明，牛单核和双核滋养层细胞在体外产生雌二醇和雌酮。皮质醇、孕酮和孕烯醇酮(10(-7)M)不影响雌二醇或雌酮的产生；外源性睾酮(10(-7)M)通过双核细胞促进雌二醇的产生，而不是单核细胞。我们的结果表明，从中期妊娠牛胎盘分离的滋养层细胞具有芳香化酶的功能，而不是17α-羟基酶的类固醇合成途径。
The bovine placenta has long been known as a source of steroid hormones. We performed three experiments to compare production of estrogens by bovine mononucleate and binucleate trophoblastic cells and examined effects of cortisol, progesterone, pregnenolone, testosterone, and androstenedione. In the first experiment, binucleate trophoblastic cells were purified by unit gravity sedimentation from six enzymatically dispersed placentas between 150 and 180 days of gestation. Cells (8 x 10(5)/ml) were incubated first at 37 degrees C for 6 h with Medium 199 alone (M199/6h) or with 10(-7) M cortisol (cortisol/6h). Medium then was replaced with 10(-7) M progesterone, 10(-7) M pregnenolone, 10(-7) M testosterone, or M199, and a second incubation was conducted for 4 h. Estradiol production did not differ between cells incubated for the first 6 h in M199 vs. cortisol and was not affected by progesterone or pregnenolone. Testosterone increased (p < 0.05) estradiol production. Estrone production did not differ between cells incubated for the first 6 h in M199 vs. cortisol; estrone production was not affected by either progesterone, pregnenolone, or testosterone.Mononucleate as well as binucleate cells were purified from placentas between 165 and 180 days of gestation and used in two other experiments. In the first of these, enriched populations of binucleate and mononucleate cells were incubated first for 6 h with Medium 199 (M199) or 10(-7) M cortisol. Medium then was replaced with 10(-7) M testosterone, 10(-7) M androstenedione, or M199 and incubation continued for 4 h. Mononucleate cells produced more estrone than did binucleate cells during the first G-h incubation (p < 0.05); cortisol did not affect this production. Estrone production by mononucleate cells was not affected by testosterone or androstenedione. Estradiol production by cells incubated for the first 6 h was not different between treatments or cell types. Testosterone increased estradiol production by binucleate cells (P < 0.05), but not mononucleate cells. In the third in this series of experiments, binucleate and mononucleate cells were incubated for 10 continuous hours with M199, progesterone, or pregnenolone. Estrone production by mononucleate cells was not affected by progesterone or pregnenolone. However, estrone production was greater by mononucleate than by binucleate cells (p < 0.05). Estradiol production was not different among treatments or cell types. These data show that bovine mononucleate and binucleate trophoblastic cells produced estradiol and estrone in vitro. Cortisol, progesterone, and pregnenolone (10(-7) M) did not affect estradiol or estrone production; and exogenous testosterone (10(-7) M) facilitated estradiol production by binucleate, but not mononucleate, cells. Our results suggest that the aromatase, but not the 17 alpha-hydroxylase, steroidogenic pathway is functional in trophoblastic cells isolated from second trimester bovine placentas.