Swainsonine differentially affects steroidogenesis and viability in caprine luteal cells in vitro
Swainsonine differentially affects steroidogenesis and viability in caprine luteal cells in vitro
复制标题
苦马豆素对体外山羊黄体细胞的类固醇生成和活力有不同影响
DOI：
10.1016/j.theriogenology.2013.03.007
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发表时间：
2013-07-01
期刊：
影响因子：
2.8
通讯作者：
Tong, Dewen
中科院分区：
文献类型：
--
作者：
Huang, Yong;Li, Wei;Tong, Dewen
据报道，家畜长期摄入含有苦马豆素 (SW) 的植物会损害生殖功能和生育能力。然而，SW 对黄体细胞类固醇生成的直接影响仍不清楚。在本研究中，使用原代和转染的黄体细胞来研究 SW 对孕酮分泌和细胞活力的影响以及这些过程中涉及的机制。用不同浓度的SW处理24或48小时后，分别使用放射免疫测定法和台盼蓝染料排除测定法评估孕酮产生和活细胞数量。较低浓度的 SW 增强基础、22R-羟基胆固醇或孕烯醇酮刺激的孕酮分泌 (P < 0.05)，而较高浓度的 SW 抑制孕酮分泌 (P < 0.05)。较低浓度的SW可促进P450侧链裂解酶和3β-羟基类固醇脱氢酶（参与黄体细胞类固醇生成的两种关键酶）在mRNA和蛋白质水平上的表达（P < 0.05），但不影响类固醇生成急性调节蛋白的表达和细胞增殖。相反，较高浓度的SW通过诱导生长期1/静止状态细胞周期停滞和细胞凋亡来抑制黄体细胞增殖（P < 0.05）。总而言之，这些结果表明，较低浓度的 SW 通过上调 P450 侧链裂解酶和 3 β-羟基类固醇脱氢酶诱导孕酮产生，而不影响细胞活力，而较高浓度的 SW 则诱导细胞周期停滞和细胞凋亡，并损害类固醇生成。这些发现为理解 SW 对黄体细胞类固醇生成的影响提供了新的见解。 (c) 2013 Elsevier Inc. 保留所有权利。
Plants containing swainsonine (SW) have been reported to impair reproductive function and fertility after long-term ingestion by livestock. However, direct effects of SW on luteal cell steroidogenesis remain unclear. In this study, primary and transfected luteal cells were used to investigate the effects of SW on progesterone secretion and cell viability and the mechanisms involved in these processes. After treatment with various concentrations of SW for 24 or 48 hours, progesterone production and the number of living cells were assessed using radioimmunoassay and trypan blue dye exclusion assay, respectively. Lower concentrations of SW enhanced basal, 22R-hydroxycholesterol- or pregnenolone-stimulated progesterone secretion (P < 0.05), whereas higher concentrations of SW inhibited progesterone secretion (P < 0.05). Lower concentrations of SW promoted expression of P450 side-chain cleavage enzyme and 3 beta-hydroxysteroid dehydrogenase, two key enzymes involved in luteal cell steroidogenesis, at mRNA and protein levels (P < 0.05), but did not affect expression of steroidogenic acute regulatory protein and cell proliferation. In contrast, higher concentrations of SW inhibited luteal cell proliferation by inducing growth phase 1/quiescent state cell cycle arrest and apoptosis (P < 0.05). Taken together, these results demonstrated that lower concentrations of SW induced progesterone production through upregulation of P450 side-chain cleavage enzyme and 3 beta-hydroxysteroid dehydrogenase without affecting cell viability, whereas higher concentrations of SW induced cell cycle arrest and apoptosis and impaired steroidogenesis. These findings provided new insights into understanding the effect of SW on luteal cell steroidogenesis. (c) 2013 Elsevier Inc. All rights reserved.