Garraza Marisa Hilda, Forneris Myriam, Virginia Gatica Laura, Oliveros Liliana Beatriz
Laboratorio de Biología de la Reproducción (LABIR), Facultad de Química, Bioquímica y Farmacia, Universidad Nacional de San Luis. San Luis, Argentina.
Neuro Endocrinol Lett. 2010;31(1):103-12.
Ovarian steroids are modulated by neural influences. In this work we investigate whether norepinephrine (NE) modifies the vasoactive intestinal peptide (VIP) or neuropeptide Y (NPY) actions in coeliac ganglion (CG) on the ovarian hormone release, and evaluate the participation of nitric oxide (NO), measured as nitrite, and of inducible nitric oxide synthetase (iNOS) protein, nerve growth factor (NGF) and its trkA receptor gene expression in the ovarian response.
The study was performed in the ex vivo CG-superior ovarian nerve (SON)-ovary system of rats on diestrus day 2 (D2). CG and ovary were placed in separate compartments connected by the SON and incubated with Krebs-Ringer buffer. After addition of 50 ng/ml VIP, 50 ng/ml NPY, 10-6 M NE, or a mix of VIP+NE or NPY+NE in ganglion, samples from the ovarian compartment were taken at different times throughout 180 minutes to measure progesterone, androstenedione and nitrite levels.
VIP and NPY in ganglion induced an increase of progesterone release that was associated for VIP, but not NPY, with a decrease of ovarian nitrite levels, iNOS protein, and NGF/trkA receptor mRNA expression. By contrast, NE in ganglion decreased progesterone, an effect that was suppressed by addition of propranolol in ganglion, and increased nitrites/iNOS and NGF/trkA receptor expression in ovary. GABA A receptor antagonist bicuculline (20 muM) added in ovarian compartment prevented the inhibitory effect on progesterone caused by NE in CG. Androstenedione was not modified under neuropeptides or NE ganglionic stimulation.
Finally, results from VIP+NE or NPY+NE in ganglion showed that ovarian response on D2 induced by VIP or NPY alone is moderated by the opposite action of NE, and occurs only on progesterone, the most sensitive steroid to neural action.
卵巢甾体激素受神经影响的调节。在本研究中,我们探究去甲肾上腺素(NE)是否会改变腹腔神经节(CG)中血管活性肠肽(VIP)或神经肽Y(NPY)对卵巢激素释放的作用,并评估以亚硝酸盐形式测定的一氧化氮(NO)、诱导型一氧化氮合酶(iNOS)蛋白、神经生长因子(NGF)及其trkA受体基因表达在卵巢反应中的参与情况。
该研究在处于动情周期第2天(D2)的大鼠离体CG - 卵巢上神经(SON) - 卵巢系统中进行。将CG和卵巢置于通过SON连接的单独隔室中,并用 Krebs - Ringer缓冲液孵育。在神经节中添加50 ng/ml VIP、50 ng/ml NPY、10 - 6 M NE或VIP + NE或NPY + NE的混合物后,在180分钟内的不同时间从卵巢隔室采集样本,以测量孕酮、雄烯二酮和亚硝酸盐水平。
神经节中的VIP和NPY诱导孕酮释放增加,对于VIP而言,这与卵巢亚硝酸盐水平、iNOS蛋白以及NGF/trkA受体mRNA表达的降低相关,但NPY则不然。相比之下,神经节中的NE降低了孕酮水平,在神经节中添加普萘洛尔可抑制该作用,并且NE增加了卵巢中亚硝酸盐/iNOS以及NGF/trkA受体的表达。在卵巢隔室中添加GABAA受体拮抗剂荷包牡丹碱(20 μM)可防止CG中的NE对孕酮产生的抑制作用。在神经肽或NE神经节刺激下,雄烯二酮未发生改变。
最后,神经节中VIP + NE或NPY + NE的结果表明,VIP或NPY单独诱导的D2期卵巢反应会受到NE相反作用的调节,并且仅发生在孕酮上,孕酮是对神经作用最敏感的甾体激素。