Grasselli Francesca, Bussolati Simona, Grolli Stefano, Di Lecce Rosanna, Dall'Aglio Cecilia, Basini Giuseppina
Dipartimento di Scienze Medico-Veterinarie, Università degli Studi di Parma, Via del Taglio 10, 43126 Parma, Italy.
Dipartimento di Medicina Veterinaria, Università degli Studi di Perugia, Via San Costanzo 4, 06126 Perugia, Italy.
Animals (Basel). 2021 Jun 17;11(6):1812. doi: 10.3390/ani11061812.
In addition to the well-known central modulatory role of orexins, we recently demonstrated a peripheral involvement in swine granulosa cells for orexin A and in adipose tissue for orexin B (OXB). The aim of present research was to verify immunolocalization of OXB and its potential role in modulating the main features of swine granulosa cells. In particular, we explored the effects on granulosa cell proliferation (through the incorporation of bromodeoxyuridine), cell metabolic activity (as indirect evaluation by the assessment of ATP), steroidogenic activity (by immunoenzymatic examination) and redox status (evaluating the production of superoxide anion by means of the WST test, production of nitric oxide through the use of the Griess test and the non-enzymatic reducing power by FRAP test). Our data point out that OXB does not modify granulosa cell growth, steroidogenesis and superoxide anion generation. On the contrary, the peptide stimulates ( < 0.05) nitric oxide output and non-enzymatic reducing power. Since new vessel growth is crucial for ovarian follicle development, a further aim of this study was to explore the expression of prepro-orexin and the effects of OXB on swine aortic endothelial cells. We found that the peptide is ineffective in modulating cell growth, while it inhibits redox status parameters. In addition, we demonstrated a stimulatory effect on angiogenesis evaluated in fibrin gel angiogenesis assay. Taken together, OXB appears to be potentially involved in the modulation of redox status in granulosa and endothelial cells and we could argue an involvement of the peptide in the follicular angiogenic events.
除了众所周知的食欲素的中枢调节作用外，我们最近还证明了食欲素A在猪颗粒细胞中的外周作用以及食欲素B（OXB）在脂肪组织中的外周作用。本研究的目的是验证OXB的免疫定位及其在调节猪颗粒细胞主要特征方面的潜在作用。具体而言，我们探讨了其对颗粒细胞增殖（通过溴脱氧尿苷掺入法）、细胞代谢活性（通过评估ATP进行间接评估）、类固醇生成活性（通过免疫酶检测）和氧化还原状态（通过WST试验评估超氧阴离子的产生、通过Griess试验评估一氧化氮的产生以及通过FRAP试验评估非酶还原能力）的影响。我们的数据表明，OXB不会改变颗粒细胞的生长、类固醇生成和超氧阴离子的产生。相反，该肽会刺激（<0.05）一氧化氮的输出和非酶还原能力。由于新血管生长对卵泡发育至关重要，本研究的另一个目的是探索前食欲素原的表达以及OXB对猪主动脉内皮细胞的影响。我们发现该肽在调节细胞生长方面无效，而它会抑制氧化还原状态参数。此外，我们在纤维蛋白凝胶血管生成试验中证明了其对血管生成的刺激作用。综上所述，OXB似乎可能参与了颗粒细胞和内皮细胞氧化还原状态的调节，并且我们可以认为该肽参与了卵泡血管生成事件。
