Miceli Fiorella, Minici Francesca, Tropea Anna, Catino Stefania, Orlando Mariateresa, Lamanna Giuseppina, Sagnella Francesca, Tiberi Federica, Bompiani Adriano, Mancuso Salvatore, Lanzone Antonio, Apa Rosanna
Cattedra Di Fisiopatologia Della Riproduzione Umana, Università Cattolica Del Sacro Cuore, 00168 Roma, Italy.
Biol Reprod. 2005 Mar;72(3):628-32. doi: 10.1095/biolreprod.104.032318. Epub 2004 Nov 17.
We investigated the effect of nicotine and its methylated metabolite, N-methyl-nicotine (M-nicotine), on human luteal cells by measuring release of progesterone and prostaglandins (PGs) from cultured cells and by testing gene expression of vascular endothelial growth factor (VEGF), an angiogenic factor strictly involved in luteal pathophysiology. Primary cultures of human luteal cells were treated for 24 h with nicotine and M-nicotine (from 10(-6) to 10(-11) M) either alone or combined with hCG (25 ng/ml); progesterone and PGs were assayed in the culture medium. In another group of experiments, luteal cells were treated for 24 h with nicotine and M-nicotine (10(-7) M) to perform reverse transcriptase-polymerase chain reaction on VEGF mRNA. Nicotine and M-nicotine negatively affected basal luteal steroidogenesis at all tested concentrations, but neither was able to affect hCG-induced progesterone release. Both substances were able to significantly increase PGF2alpha release from luteal cells, with a dose-related efficacy for M-nicotine. On the contrary, PGE2 release was significantly inhibited by both nicotine and its metabolite. Finally, nicotine was able to increase VEGF mRNA expression significantly, whereas M-nicotine was not. In conclusion, nicotine and M-nicotine can induce a sort of luteal insufficiency by inhibiting progesterone release, probably through modulation of the PG system.
我们通过检测培养细胞中孕酮和前列腺素（PGs）的释放，并检测血管内皮生长因子（VEGF）的基因表达，研究了尼古丁及其甲基化代谢产物N-甲基尼古丁（M-尼古丁）对人黄体细胞的影响。VEGF是一种严格参与黄体病理生理过程的血管生成因子。人黄体细胞原代培养物单独或与hCG（25 ng/ml）联合用尼古丁和M-尼古丁（浓度从10⁻⁶到10⁻¹¹ M）处理24小时；测定培养基中的孕酮和PGs。在另一组实验中，黄体细胞用尼古丁和M-尼古丁（10⁻⁷ M）处理24小时，以对VEGF mRNA进行逆转录聚合酶链反应。在所有测试浓度下，尼古丁和M-尼古丁均对基础黄体类固醇生成产生负面影响，但两者均不能影响hCG诱导的孕酮释放。两种物质均能显著增加黄体细胞中PGF2α的释放，M-尼古丁具有剂量相关效应。相反，尼古丁及其代谢产物均显著抑制PGE2的释放。最后，尼古丁能够显著增加VEGF mRNA的表达，而M-尼古丁则不能。总之，尼古丁和M-尼古丁可能通过调节PG系统抑制孕酮释放，从而诱导一种黄体功能不全。
