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前列腺素F(2α)调节牛黄体内皮细胞中的一氧化氮生成系统。

Prostaglandin F(2alpha) regulates the nitric oxide generating system in bovine luteal endothelial cells.

作者信息

Lee Seung-Hyung, Acosta Tomas J, Yoshioka Shin, Okuda Kiyoshi

机构信息

Laboratory of Reproductive Endocrinology, Graduate School of Natural Science and Technology, Okayama University, Okayama, Japan.

出版信息

J Reprod Dev. 2009 Aug;55(4):418-24. doi: 10.1262/jrd.20205. Epub 2009 Apr 30.

Abstract

The objective of the present study was to elucidate whether luteolytic prostaglandin F(2alpha) (PGF) plays roles in regulating the nitric oxide (NO) generating system in luteal endothelial cells (LECs). Reverse transcriptase PCR, immunoblotting and immunostaining revealed the presence of PGF receptor mRNA (521 bp) and protein (64 kDa) in cultured LECs obtained from the mid-stage corpus luteum. When cultured LECs were exposed to 0.1 microM-10 microM PGF, NO production was significantly stimulated by PGF at 24 h. When LECs were exposed to 1 microM PGF for 2, 6 and 24 h, PGF did not affect the expressions of endothelial NO synthase (eNOS) mRNA and protein. On the other hand, PGF stimulated the expression of inducible NOS (iNOS) mRNA (P<0.05) and protein (P<0.05) at 2 h, but not at 6 and 24 h. By observing the conversion of (3)C-arginine to (3)C-citrulline, we found that PGF stimulated NOS activity in cultured LECs at 2 h (P<0.05). The overall findings indicate that bovine LECs are a target for PGF and that PGF stimulates iNOS expression and NOS activity in bovine LECs. Stimulation of the NO generating system and NOS activity by PGF may result in increasing local NO production followed by luteolysis.

摘要

本研究的目的是阐明黄体溶解前列腺素F(2α)(PGF)是否在调节黄体内皮细胞(LECs)中的一氧化氮(NO)生成系统中发挥作用。逆转录聚合酶链反应、免疫印迹和免疫染色显示,从黄体中期获得的培养LECs中存在PGF受体mRNA(521 bp)和蛋白(64 kDa)。当培养的LECs暴露于0.1微摩尔/升 - 10微摩尔/升的PGF时,PGF在24小时时显著刺激了NO的产生。当LECs暴露于1微摩尔/升的PGF 2、6和24小时时,PGF不影响内皮型一氧化氮合酶(eNOS)mRNA和蛋白的表达。另一方面,PGF在2小时时刺激了诱导型一氧化氮合酶(iNOS)mRNA(P<0.05)和蛋白(P<0.05)的表达,但在6和24小时时没有。通过观察(3)C-精氨酸向(3)C-瓜氨酸的转化,我们发现PGF在2小时时刺激了培养LECs中的一氧化氮合酶活性(P<0.05)。总体研究结果表明,牛LECs是PGF的靶细胞,并且PGF刺激牛LECs中的iNOS表达和一氧化氮合酶活性。PGF对NO生成系统和一氧化氮合酶活性的刺激可能导致局部NO产生增加,随后黄体溶解。

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