Sirotkin A V, Makarevich A V, Kwon H B, Kotwica J, Bulla J, Hetényi L
Research Institute of Animal Production, 949 92 Nitra, Slovakia.
J Endocrinol. 2001 Dec;171(3):475-80. doi: 10.1677/joe.0.1710475.
The aims of this study on porcine ovarian granulosa cells were to examine the effect of GH on oxytocin (OT), IGF-I and IGF-I receptors, IGF-binding protein-3 (IGFBP-3), progesterone and prostaglandin E (PGE), as well as to determine whether IGF-I and/or OT may be mediators of GH action. The cells were cultured either with porcine GH (pGH) (1 ng/ml to 10 microg/ml or 100 ng/ml only), antiserum against IGF-I (0.1%), antiserum against OT (0.1%) or a combination of GH (10 ng/ml) with antiserum against IGF-I or antiserum against OT (0.1%). The secretion of IGF-I, OT, IGFBP-3, progesterone and PGE was determined using RIA/IRMA, whilst the IGF-I binding sites were measured using a radioreceptor assay. It was observed that pGH increased the secretion of IGF-I and the abundance of IGF-I binding sites in granulosa cells. Furthermore, GH inhibited OT release, stimulated progesterone and PGE output, but had no significant effect on IGFBP-3 secretion. Immunoneutralization of IGF-I by antiserum against IGF-I inhibited PGE secretion, but it did not influence progesterone or IGFBP-3 secretion. Binding of OT by antiserum suppressed IGFBP-3, PGE, but not progesterone secretion. Neither immunoneutralization of IGF-I nor OT substantially prevented the effects of GH on progesterone, IGFBP and PGE. These observations demonstrate the involvement of GH, IGF-I and OT in the control of porcine ovarian secretory activity and the ability of GH to regulate IGF-I and OT production and IGF-I reception. Nevertheless, lack of correlation between the effects of GH, antiserum against IGF-I and antiserum against OT, as well as the inability of blockade of IGF-I or OT to prevent the effects of GH, suggests that IGF-I and OT, despite their dependence on GH, do not mediate GH action on ovarian cells.
本研究以猪卵巢颗粒细胞为对象,旨在检测生长激素(GH)对催产素(OT)、胰岛素样生长因子-I(IGF-I)及其受体、胰岛素样生长因子结合蛋白-3(IGFBP-3)、孕酮和前列腺素E(PGE)的影响,并确定IGF-I和/或OT是否可能是GH作用的介质。细胞分别用猪GH(pGH)(1 ng/ml至10 μg/ml或仅100 ng/ml)、抗IGF-I抗血清(0.1%)、抗OT抗血清(0.1%)或GH(10 ng/ml)与抗IGF-I抗血清或抗OT抗血清(0.1%)的组合进行培养。采用放射免疫分析/免疫放射分析测定IGF-I、OT、IGFBP-3、孕酮和PGE的分泌,同时用放射受体分析法测量IGF-I结合位点。观察到pGH增加了颗粒细胞中IGF-I的分泌和IGF-I结合位点的丰度。此外,GH抑制OT释放,刺激孕酮和PGE的分泌,但对IGFBP-3的分泌无显著影响。抗IGF-I抗血清对IGF-I的免疫中和作用抑制了PGE的分泌,但不影响孕酮或IGFBP-3的分泌。抗血清对OT的结合抑制了IGFBP-3、PGE的分泌,但不影响孕酮的分泌。IGF-I或OT的免疫中和均未实质性地阻止GH对孕酮、IGFBP和PGE的作用。这些观察结果表明GH、IGF-I和OT参与了猪卵巢分泌活动的调控,以及GH调节IGF-I和OT产生及IGF-I受体的能力。然而,GH、抗IGF-I抗血清和抗OT抗血清的作用之间缺乏相关性,以及阻断IGF-I或OT无法阻止GH的作用,这表明IGF-I和OT尽管依赖于GH,但并不介导GH对卵巢细胞的作用。