Inhibin production by primary Sertoli cell-enriched cultures: regulation by follicle-stimulating hormone, androgens, and epidermal growth factor.
Inhibin production by primary Sertoli cell-enriched cultures: regulation by follicle-stimulating hormone, androgens, and epidermal growth factor.
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DOI：
10.1210/endo-122-2-717
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发表时间：
1988-02
期刊：
影响因子：
4.8
通讯作者：
PATRICIA L. Morris;W. Vale;Scott Cappel;C. W. Bardin
中科院分区：
文献类型：
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作者：
PATRICIA L. Morris;W. Vale;Scott Cappel;C. W. Bardin
研究了原代Sertoli细胞富集型(SCE)细胞培养上清液中抑制素水平随时间和激素处理的变化。取20日龄大鼠建立姐妹染色单体交换培养体系，在添加胰岛素、转铁蛋白、表皮生长因子(EGF)和杆菌素的SF培养液中培养。用针对合成的猪抑制素-α(PIα)的抗体测定可放射免疫测定的抑制素，并用同样的合成肽和高纯度的绵羊抑制素标准进行测定。这些测定的结果以合成肽的形式表示，即PIα-(1-26)-G-Y-每毫升/10(6)细胞的飞摩尔。对照组培养上清液中以此速率累积的抑制素水平(+/-扫描电子显微镜)在0-4天和4-8天分别为184.9+/-6.1和167.4+/-5.2。当加入FSH(oFSH 17；1-1000 ng/ml)时，抑制素水平在初始24 h后的所有时间点均呈剂量依赖性升高，2×10(-7)M睾酮(T)与低剂量FSH同时加入抑制素(T)抑制FSH的反应，但当T与300 ng/mlFSH联合作用时，与单独FSH相比，T对抑制素水平无明显影响。尽管T的作用不大，雄烯二酮(A；2×10(-13)至x10(-5)M)对抑制素水平有剂量依赖性的抑制作用。在所有剂量的FSH中也观察到这种抑制作用。A的作用不是由于其转化为雌激素，因为在有或没有FSH的情况下，17β-雌二醇对姐妹染色单体交换培养物抑制素的产生都没有影响。接下来研究了无血清基础培养液中的表皮生长因子的作用；它产生的抑制素水平(188.7+/-9.5)是不含表皮生长因子的培养物(135.6+/-5.0)的1.5倍。当加入FSH和EGF时，抑制素水平的刺激是相加的。我们的结论是：(1)在未成熟大鼠的支持细胞培养中，(1)培养液中抑制素的积累量从最初的90fmol/10(6)细胞·天(最初的24小时)下降到40-50 fmol/10(6)细胞·天(最初的48小时)，并在培养8天内持续积累；(2)FSH调节支持细胞的抑制素的产生，3天的剂量反应最好。
Inhibin levels in the serum-free media of primary Sertoli cell-enriched (SCE) cultures were studied as a function of time and hormonal treatment. SCE cultures were established from 20-day-old rats and maintained in SF media supplemented with insulin, transferrin, epidermal growth factor (EGF), and bacitracin. Radioimmunoassayable inhibin was measured using an antibody directed against a synthetic porcine inhibin-alpha (pI alpha) and measured using this same synthetic peptide as well as a highly purified ovine inhibin standard. Results of these determinations are expressed in terms of a synthetic peptide as femtomoles of pI alpha-(1-26)-G-Y-per ml/10(6) cells. Inhibin levels (+/- SEM) that accumulated at this rate in media from control cultures were 184.9 +/- 6.1 and 167.4 +/- 5.2 on days 0-4 and 4-8, respectively. When FSH (oFSH 17; 1-1000 ng/ml) was added, a dose-dependent increase in inhibin levels was significant at all time points beyond the initial 24 h. The simultaneous addition of 2 x 10(-7) M testosterone (T) with low doses of FSH addition of 2 x 10(-7) m testosterone (T) with low doses of FSH suppressed the inhibin response to FSH, but when T was combined with 300 ng/ml FSH, there was no effect of T on inhibin levels compared to FSH alone, regardless of time in culture. In spite of the modest effect of T, androstenedione (A; 2 x 10(-13) to x 10(-5) M) produced a dose-dependent suppression of inhibin levels. This inhibition was also observed at all doses of FSH. The action of A was not due to its conversion to estrogens, as 17 beta-estradiol had no effect on inhibin production by SCE cultures in either the presence or absence of FSH. The effect of EGF, a component of the basal serum-free medium, was next examined; it produced a 1.5-fold higher level of inhibin (188.7 +/- 9.5) compared to cultures without EGF (135.6 +/- 5.0). When SCE cultures were plated with FSH plus EGF, the stimulation of inhibin levels was additive. We conclude that in Sertoli cell cultures established from immature rats (1) the accumulation of inhibin in medium declines from 90 fmol/10(6) cells.day (initial 24 h) to 40-50 fmol/10(6) cells.day (over the first 48 h) and continues to accumulate in the medium for 8 days of culture; (2) FSH regulates the production of inhibin by Sertoli cells; the best dose response is observed over a 3-day period.(ABSTRACT TRUNCATED AT 400 WORDS)