Melatonin Exerts Direct Inhibitory Actions on ACTH Responses in the Human Adrenal Gland
Melatonin Exerts Direct Inhibitory Actions on ACTH Responses in the Human Adrenal Gland
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DOI：
10.1055/s-0031-1271693
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发表时间：
2011-05-01
影响因子：
2.2
通讯作者：
Seron-Ferre, M.
中科院分区：
文献类型：
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作者：
Campino, C.;Valenzuela, F. J.;Seron-Ferre, M.
在非人灵长类动物和啮齿动物中，褪黑素直接作用于肾上腺，抑制糖皮质激素对ACTH的反应。在这些物种中，一个内在的肾上腺生物钟参与促肾上腺皮质激素刺激的糖皮质激素的产生。我们通过测定i)体内生物钟基因的表达，ii)褪黑激素对acth刺激的肾上腺外植体的直接影响，以及a)生物钟基因PER1（周期1）mRNA和BMAL1[脑肌（ARNT）样]蛋白、acth诱导的类固醇急性调节蛋白（StAR）和3 β -羟基类固醇脱氢酶（3 β - hsd）的表达，以及b)对皮质醇和黄体酮产生的影响，来研究这些发现是否适用于人肾上腺。6例单侧肾切除-肾上腺切除术的肾癌患者的肾上腺组织。采用RT-PCR方法研究了正常肾上腺组织和腺瘤组织中生物钟基因PER1、PER2、CRY2、clock和BMAL1的表达。在独立实验中，4个正常肾上腺的外植体在培养基中预孵育（6 h），然后在单独培养基中孵育12 h；ACTH (100 nM)；ACTH加褪黑素（100 nM）；还有褪黑素。测定外植体PER1 mRNA （real-time PCR）和StAR、3 β - hsd、BMAL1（免疫槽点印迹法）的含量及其皮质醇和孕酮的分泌量（RIA）。人类肾上腺表达时钟基因PER1、PER2、CRY2、clock和BMAL1。ACTH增加PER1 mRNA、BMAL1、StAR和3 β - hsd蛋白水平，以及皮质醇和黄体酮的产生。褪黑素抑制ACTH的作用。我们的研究首次证明了褪黑素对人肾上腺中几种ACTH反应的直接抑制作用。
In nonhuman primates and rodents, melatonin acting directly on the adrenal gland, inhibits glucocorticoid response to ACTH. In these species, an intrinsic adrenal circadian clock is involved in ACTH-stimulated glucocorticoid production. We investigated whether these findings apply to the human adrenal gland by determining i) expression of clock genes in vivo and ii) direct effects of melatonin in ACTH-stimulated adrenal explants over a) expression of the clock genes PER1 (Period 1) mRNA and BMAL1 [Brain-Muscle (ARNT)like] protein, ACTH-induced steroidogenic acute regulatory protein (StAR), and 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) and b) over cortisol and progesterone production. Adrenal tissue was obtained from 6 renal cancer patients undergoing unilateral nephrectomy-adrenalectomy. Expression of the clock genes PER1, PER2, CRY2 (Cryptochrome 2), CLOCK (Circadian Locomotor Output Cycles Kaput) and BMAL1, was investigated by RT-PCR in a normal adrenal and in an adenoma. In independent experiments, explants from 4 normal adrenals were preincubated in culture medium (6 h) followed by 12 h in: medium alone; ACTH (100 nM); ACTH plus melatonin (100 nM); and melatonin alone. The explants' content of PER1 mRNA (real-time PCR) and StAR, 3 beta-HSD, BMAL1 (immuno slot-blot), and their cortisol and progesterone production (RIA) were measured. The human adrenal gland expresses the clock genes PER1, PER2, CRY2, CLOCK, and BMAL1. ACTH increased PER1 mRNA, BMAL1, StAR, and 3 beta-HSD protein levels, and cortisol and progesterone production. Melatonin inhibited these ACTH effects. Our study demonstrates, for the first time, direct inhibitory effects of melatonin upon several ACTH responses in the human adrenal gland.