Department of Medicine and Clinical Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama 700-8558, Japan.
Mol Cell Endocrinol. 2011 Sep 15;344(1-2):41-50. doi: 10.1016/j.mce.2011.06.016. Epub 2011 Jul 1.
Mechanisms by which GHRP stimulates ACTH release in corticotrope cells were investigated using mouse corticotrope AtT20 cells by focusing on the biological activity of BMP-4. GHRP-2 increased ACTH and cAMP secretion by AtT20 cells; however, its effects were less potent than the effects of CRH. BMP-4 suppressed basal ACTH production and POMC transcription, and the inhibition of endogenous BMP receptor signaling led to an increase in ACTH production. Of note, BMP-4 suppressed ACTH production and POMC-promoter activity induced by CRH more efficaciously than that induced by GHRP-2. BMP-4 had no significant effect on cAMP synthesis induced by CRH or GHRP-2. Stimulation with CRH, but not GHRP-2, activated ERK1/2, p38, SAPK/JNK and Akt phosphorylation, in which CRH-induced phosphorylation of ERK and p38 was suppressed by BMP-4. GHRP-2-induced ACTH secretion was not affected by inhibitors of ERK, p38 and Akt pathways, which effectively suppressed CRH-induced ACTH release. Blockage of the cAMP-PKA pathway reversed CRH- as well as GHRP-2-induced ACTH secretion. Furthermore, the inhibition of ERK and p38 significantly reduced cAMP synthesis induced by CRH but not by GHRP-2. Thus, CRH activates ACTH production through ERK and p38 pathways in addition to the cAMP-PKA pathway, which is also activated downstream of MAPK. On the other hand, GHRP-2-induced ACTH production was predominantly linked to the cAMP-PKA pathway. Moreover, CRH and GHRP-2 upregulated BMP receptor signaling, while BMP-4, CRH and GHRP-2 had no significant effect on the expression level of GHSR. In addition, GHRP-2 suppressed the expression of Smad7, which is an inhibitor of the BMP-Smad1/5/8 pathway. Collectively, the results revealed a functional interaction between GHRP-2 and BMP signaling, in which endogenous BMP may act as an autoregulatory system in controlling ACTH production.
我们通过研究小鼠 corticotrope AtT20 细胞,聚焦于 BMP-4 的生物学活性,来研究 GHRP 刺激促肾上腺皮质激素释放细胞中 ACTH 释放的机制。GHRP-2 增加了 AtT20 细胞中 ACTH 和 cAMP 的分泌;然而,其作用不如 CRH 强。BMP-4 抑制基础 ACTH 的产生和 POMC 转录,并且抑制内源性 BMP 受体信号导致 ACTH 的产生增加。值得注意的是,BMP-4 抑制 CRH 诱导的 ACTH 产生和 POMC 启动子活性的作用比 GHRP-2 更有效。BMP-4 对 CRH 或 GHRP-2 诱导的 cAMP 合成没有显著影响。CRH 刺激而非 GHRP-2 刺激激活了 ERK1/2、p38、SAPK/JNK 和 Akt 磷酸化,其中 BMP-4 抑制了 CRH 诱导的 ERK 和 p38 的磷酸化。ERK、p38 和 Akt 通路抑制剂对 GHRP-2 诱导的 ACTH 分泌没有影响,而这些抑制剂有效地抑制了 CRH 诱导的 ACTH 释放。阻断 cAMP-PKA 通路逆转了 CRH 和 GHRP-2 诱导的 ACTH 分泌。此外,ERK 和 p38 的抑制显著减少了 CRH 诱导但不 GHRP-2 诱导的 cAMP 合成。因此,CRH 通过 ERK 和 p38 通路以及 cAMP-PKA 通路激活 ACTH 产生,后者也被 MAPK 下游激活。另一方面,GHRP-2 诱导的 ACTH 产生主要与 cAMP-PKA 通路有关。此外,CRH 和 GHRP-2 上调了 BMP 受体信号,而 BMP-4、CRH 和 GHRP-2 对 GHSR 的表达水平没有显著影响。此外,GHRP-2 抑制了 BMP-Smad1/5/8 通路抑制剂 Smad7 的表达。总的来说,这些结果揭示了 GHRP-2 和 BMP 信号之间的功能相互作用,其中内源性 BMP 可能作为控制 ACTH 产生的自身调节系统发挥作用。