Effect of bacterial endotoxin lipopolysaccharide treatment on duck Leydig cells
Effect of bacterial endotoxin lipopolysaccharide treatment on duck Leydig cells
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DOI：
10.21451/1984-3143-ar2019-0002
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发表时间：
2019-10-01
影响因子：
1.7
通讯作者：
Huang, Yunmao
中科院分区：
文献类型：
--
作者：
Lao, Yongcong;Ouyang, Hongjia;Huang, Yunmao
本研究旨在探讨细菌内毒素脂多糖（LPS）对鸭间质细胞激素产生和基因表达的影响及其机制。采集200日龄绿头鸭的间质细胞，分为5个处理组（0、50、100、200和400 ng/mL LPS）。LPS处理6、12、24和48小时后，使用酶联免疫吸附试验（ELISA）试剂盒检测细胞上清液中的睾酮、激活素和抑制素水平。采用实时定量聚合酶链式反应（qRT-PCR）检测睾酮合成相关基因（甾体急性调节蛋白（StAR）、3- β -羟基类固醇脱氢酶（3 - β - hsd）、细胞色素P450芳香化酶（P450arom））和生殖相关基因（促性腺激素抑制激素受体（GnIHR）、促卵泡激素受体（FSHR）、促黄体生成素受体（LHR））的表达水平。我们成功地分离培养了鸭间质细胞，细胞纯度在90%以上。与对照组相比，随着LPS浓度的升高，间质细胞分泌的睾酮、激活素和抑制素水平逐渐降低。LPS处理后，间质细胞中StAR和3 β - hsd基因在12 h表达上调，GnIHR在24 h表达上调；而FSHR和LHR的表达在24 h时降低。本研究表明，LPS可以抑制鸭间质细胞的激素分泌并调节相关基因的表达。
This study aimed to investigate the effects of bacterial endotoxin lipopolysaccharide (LPS) on hormone production and gene expression in duck Leydig cells and its underlying mechanisms. Leydig cells were collected from 200-day-old mallard ducks and divided into five treatment groups (0, 50, 100, 200, and 400 ng/mL LPS). After treatment with LPS for 6, 12, 24, and 48 h, testosterone, activin, and inhibin levels in the cell supernatants were determined using enzyme-linked immunosorbent assay (ELISA) kits. The expression levels of testosterone synthesis-related genes, including steroidogenic acute regulatory protein (StAR), 3-beta-hydroxysteroid dehydrogenase (3 beta-HSD), and cytochrome P450 aromatase (P450arom), and reproductive-related genes, including gonadotropin-inhibitory hormone receptor (GnIHR), follicle stimulating hormone receptor (FSHR), and luteinizing hormone receptor (LHR) were detected using quantitative real-time polymerase chain reaction (qRT-PCR). We successfully isolated and cultured duck Leydig cells with cell purity above 90%. Compared with the control group, the levels of testosterone, activin, and inhibin secreted in Leydig cells decreased gradually with increasing LPS concentration. After treatment with LPS, the expression of StAR and 3 beta-HSD genes in Leydig cells was upregulated at 12 h, and that of GnIHR was upregulated at 24 h; whereas the expression of FSHR and LHR was reduced at 24 h. This study indicates that LPS can inhibit the secretion of hormones and regulate the expression of related genes in duck Leydig cells.