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氯化汞抑制蛋鸡卵巢颗粒细胞孕酮分泌的分子机制

Molecular mechanism of mercuric chloride inhibiting progesterone secretion in ovarian granulosa cells of laying hens.

作者信息

Ma Yan, Gong Yujie J, Xu Qianqian Q, Zou Xiaoting

机构信息

Key Laboratory of Animal Feed and Nutrition of Zhejiang Province, Feed Science Institute, College of Animal Science, Zhejiang University, Hangzhou, China.

出版信息

J Anim Physiol Anim Nutr (Berl). 2018 Dec;102(6):1533-1542. doi: 10.1111/jpn.12955. Epub 2018 Aug 25.

DOI:10.1111/jpn.12955
PMID:30144178
Abstract

This study investigated the effect of mercury (Hg) on progesterone secretion in ovarian granulosa cells of laying hens. The gene expressions of steroidogenic acute regulatory protein (StAR), cytochrome P450 cholesterol side-chain cleavage (P450scc) and 3β-hydroxysteroid dehydrogenase (3β-HSD), cyclic adenosine monophosphate (cAMP)-protein kinase A (PKA) pathway and intracellular calcium ion (Ca ) were further investigated to uncover the molecular mechanism. Results revealed that the cell viability was gradually decreased after Hg exposure from 0 to 24 hr. Besides, progesterone secretion was significantly decreased (p < 0.05) as the concentration of Hg increased from 0 to 4 μM followed by a plateau in 6 μM Hg group at 12-hr time point. Compared with 0 μM Hg group, 4 and 6 μM Hg for 48 hr had significantly decreased progesterone secretion (p < 0.05), while Hg exposure for 6 and 24 hr had no apparent effect on progesterone secretion. In addition, positive correlations occurred among intracellular progesterone, cAMP, PKA, mRNA expressions of StAR, P450scc and 3β-HSD at 12-h and 24-h time points. On the contrary, intracellular Ca level was negatively related to cAMP level at 6 time point and was negatively correlated with progesterone and PKA level at 48 time point. It could be concluded that Hg dose- and time-dependently inhibited progesterone secretion by means of attenuating cAMP-PKA signal pathway, gene expressions of StAR, P450scc and 3β-HSD and enhancing intracellular Ca in ovarian granulosa cells of laying hens.

摘要

本研究调查了汞(Hg)对蛋鸡卵巢颗粒细胞孕酮分泌的影响。进一步研究了类固醇生成急性调节蛋白(StAR)、细胞色素P450胆固醇侧链裂解酶(P450scc)和3β-羟基类固醇脱氢酶(3β-HSD)的基因表达、环磷酸腺苷(cAMP)-蛋白激酶A(PKA)信号通路及细胞内钙离子(Ca),以揭示其分子机制。结果显示,汞暴露0至24小时后细胞活力逐渐下降。此外,随着汞浓度从0增加到4μM,孕酮分泌显著降低(p<0.05),随后在12小时时间点,6μM汞组出现平台期。与0μM汞组相比,4μM和6μM汞处理48小时后孕酮分泌显著降低(p<0.05),而汞暴露6小时和24小时对孕酮分泌无明显影响。此外,在12小时和24小时时间点,细胞内孕酮、cAMP、PKA、StAR、P450scc和3β-HSD的mRNA表达之间呈正相关。相反,在6小时时间点,细胞内Ca水平与cAMP水平呈负相关,在48小时时间点与孕酮和PKA水平呈负相关。可以得出结论,汞通过减弱cAMP-PKA信号通路、StAR、P450scc和3β-HSD的基因表达以及增加蛋鸡卵巢颗粒细胞内钙离子水平,呈剂量和时间依赖性地抑制孕酮分泌。

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