固醇类物质对鹅颗粒细胞胆固醇转运相关基因表达的影响

doi:10.16036/j.issn.1000-2650.2019.01.016

摘要/Abstract

摘要： 【目的】揭示胆固醇转运相关基因在鹅不同发育阶段卵泡中的表达模式及固醇类物质对颗粒细胞上述基因表达的影响。【方法】首先应用RT-qPCR技术检测SREBP-2、STADR4、STAR和CYP11A1在产蛋高峰期鹅卵巢内不同发育阶段卵泡中的mRNA表达水平;然后分离鹅F1卵泡颗粒细胞体外培养,24 h后分别添加不同浓度胆固醇、25-羟胆固醇和洛伐他汀（胆固醇合成限速酶抑制剂）处理,最后使用MTT法检测细胞活性和RT-qPCR技术检测上述基因表达量。【结果】四个基因在不同发育阶段卵泡中的表达模式相似;高浓度胆固醇显著增加细胞活性（P<0.05）,25-羟胆固醇和洛伐他汀则降低细胞活性。随着胆固醇浓度升高,四个基因表达量均先升高后下降;25-羟胆固醇显著抑制SREBP-2和STAR的表达（P<0.05）;升高洛伐他汀浓度使SREBP-2表达量先降低后升高,与其余三个基因相反。【结论】胆固醇转运和类固醇激素合成相关基因表达量受固醇种类和浓度调节,并且与卵泡类固醇激素合成密切相关,这为揭示鹅卵泡发育机制和提高产蛋量提供了理论参考。

关键词: 卵泡发育, 颗粒细胞, 鹅, SREBP-2, STARD4, STAR, CYP11A1

Abstract: 【Objective】 The aim of this study was to reveal the expression pattern of cholesterol transport-related genes in geese follicles at different developmental stages and study the effects of severalsterols on the expression of above genes in geese granulosa cells. 【Method】 Ovarian follicular samples were collected from the maternal line of Tianfu meat geese during the period of egg laying. The mRNA levels of SREBP-2,STADR4, STAR and CYP11A1 were detected by RT-qPCR. Then, granulosa cells were isolated from geese F1 follicles and cultured in vitro. After cultured for 24 hours, cells were treated withdifferent concentrations of cholesterol, 25-hydroxycholesterol or lovastatin (an inhibitor of cholesterol synthesis rate-limiting enzyme). Finally, cell activity was assessedusing MTT assay, and qPCR was used to detect the expression levels of SREBP-2, STARD4, STAR and CYP11A1. 【Result】 Our results showed that the expression patterns of four genes were similar in different sized follicles. At the cellular level, high concentration of cholesterol significantly increased granulosa cell activity (P<0.05), whereas, both 25-hydroxycholesterol and lovastatin decreased cell activity. With the increase of cholesterol concentration, the expression levels of four genes increased at first and then decreased, and 25-hydroxycholesterol significantly inhibited the expressions of SREBP-2 and STAR (P<0.05), whileincreased lovastatin concentration, whichcaused the up-regulation and the subsequent decreasing of SREBP-2 expression, contrary to the other three genes. 【Conclusion】 Expression of both cholesterol transport- and steroid hormone synthesis-related genes in granulosa cellsweredependent on the type and concentration of sterol, and their tissue distribution patterns are closely related to follicular steroidogenesis. These results contributed to a better understanding of the mechanisms underlying avian follicle development and are conducive to the improvement of geese egg production.

Key words: follicular development, granulosa cells, geese, SREBP-2, STARD4, STAR, CYP11A1

中图分类号:

S835

荣玉静, 夏露, 胡深强, 王继文. 固醇类物质对鹅颗粒细胞胆固醇转运相关基因表达的影响[J]. 四川农业大学学报, 2019, 37(01): 103-109.

RONG Yujing, XIA Lu, HU Shenqiang, WANG Jiwen. Effects of Sterols on the Expression of Cholesterol Transporter Related Genes in Geese Granulosa Cells[J]. Journal of Sichuan Agricultural University, 2019, 37(01): 103-109.

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