Yu Guang-Min, Isobe Naoki, Maeda Teruo
Department of Bioresource Science, Graduate School of Biosphere Science, Hiroshima University, Higashi-Hiroshima 739-8528, Japan.
The Research Center for Animal Science, Hiroshima University, Higashi-Hiroshima 739-8528, Japan.
J Poult Sci. 2017 Oct 25;54(4):319-325. doi: 10.2141/jpsa.0170048.
The aim of this study was to evaluate the potential of melatonin to protect cultured granulosa cells from the harmful effects of lipopolysaccharide (LPS) in quail. Granulosa cells isolated from Japanese quails were pretreated with or without melatonin (10 or 100 g/mL) for 12 hand then incubated for 12 hin the absence or presence of 100 ng/mL LPS. The expression of pro-inflammatory cytokines and chemokine was detected by quantitative real-time PCR. The levels of oxidative stress biomarkers (dityrosine and nitrite) were determined by ELISA and the Griess reaction. Cell viability was quantified using an MTT assay. Additionally, the level of progesterone was measured by ELISA. We found that melatonin decreased LPS-induced expression of , and . In addition, melatonin increased the dityrosine level, but suppressed the nitrite level. Finally, melatonin administration increased the viability of LPS-stimulated granulosa cells . However, progesterone basal secretion was not significantly changed. These results suggest that melatonin protects cultured granulosa cells from LPS-induced inflammatory and oxidative stress damage and provide evidence that melatonin might have therapeutic utility in ovarian follicle infection in Japanese quail.
本研究的目的是评估褪黑素保护鹌鹑培养的颗粒细胞免受脂多糖（LPS）有害影响的潜力。从日本鹌鹑分离的颗粒细胞在有或没有褪黑素（10或100μg/mL）的情况下预处理12小时，然后在不存在或存在100 ng/mL LPS的情况下孵育12小时。通过定量实时PCR检测促炎细胞因子和趋化因子的表达。通过ELISA和格里斯反应测定氧化应激生物标志物（二酪氨酸和亚硝酸盐）的水平。使用MTT法对细胞活力进行定量。此外，通过ELISA测量孕酮水平。我们发现褪黑素降低了LPS诱导的……的表达。此外，褪黑素增加了二酪氨酸水平，但抑制了亚硝酸盐水平。最后，给予褪黑素增加了LPS刺激的颗粒细胞的活力。然而，孕酮基础分泌没有显著变化。这些结果表明，褪黑素保护培养的颗粒细胞免受LPS诱导的炎症和氧化应激损伤，并提供证据表明褪黑素可能对日本鹌鹑卵巢卵泡感染具有治疗作用。
