Rossi Gianna, Gasperi Valeria, Paro Rita, Barsacchi Daniela, Cecconi Sandra, Maccarrone Mauro
Department of Biomedical Sciences, University of Teramo, Piazza A. Moro 45, 64100 Teramo, Italy.
Endocrinology. 2007 Mar;148(3):1431-9. doi: 10.1210/en.2006-0969. Epub 2006 Nov 16.
Among the biological activities of the endocannabinoid anandamide (N-arachidonoylethanolamine) (AEA), growing interest has been attracted by the regulation of mammalian fertility. Recently we have shown that treatment of mouse primary Sertoli cells with FSH enhances the activity of the AEA hydrolase [fatty acid amide hydrolase (FAAH)], though the molecular details were not elucidated. Here, we investigated whether FSH was also able to affect the enzymes that synthesize AEA (N-acyltransferase and N-acyl-phosphatidyl-ethanolamine-phospholipase D), the endogenous content of this endocannabinoid, and the level of the AEA-binding vanilloid receptor 1 (transient receptor potential channel vanilloid receptor subunit 1). We show that FSH enhanced FAAH activity (up to approximately 500% of the controls) and expression (up to approximately 300%), leading to a marked reduction (down to approximately 15%) of AEA content. However N-acyltransferase and N-acyl-phosphatidyl-ethanolamine-phospholipase D activity, and transient receptor potential channel vanilloid receptor subunit 1 binding were not affected. We also show that diacylglycerol lipase and monoacylglycerol lipase, which respectively synthesize and degrade 2-arachidonoyl-glycerol, were not regulated by FSH, neither was the membrane transport of this endocannabinoid. In addition, we show that FAAH stimulation by FSH was abrogated by inhibitors of protein kinase A (PKA) and cytochrome-P(450) aromatase, and was conversely mimicked by N,O'-dibutyryl cAMP and estrogen. Finally, we demonstrate that FSH protects Sertoli cells against the pro-apoptotic activity of AEA, through PKA and aromatase-dependent activation of FAAH. Altogether these data suggest that FAAH is the only target of FSH among the elements of the endocannabinoid system, and that its regulation by PKA and aromatase-dependent pathways impacts Sertoli cell proliferation.
在内源性大麻素花生四烯酸乙醇胺（N-花生四烯酰乙醇胺，AEA）的生物学活性中，其对哺乳动物生育能力的调节已引起越来越多的关注。最近我们发现，用促卵泡激素（FSH）处理小鼠原代支持细胞可增强AEA水解酶[脂肪酸酰胺水解酶（FAAH）]的活性，不过分子细节尚未阐明。在此，我们研究了FSH是否也能影响合成AEA的酶（N-酰基转移酶和N-酰基磷脂酰乙醇胺-磷脂酶D）、这种内源性大麻素的内源性含量以及AEA结合香草酸受体1（瞬时受体电位香草酸受体亚基1）的水平。我们发现，FSH增强了FAAH的活性（高达对照的约500%）和表达（高达对照的约300%），导致AEA含量显著降低（降至约15%）。然而，N-酰基转移酶和N-酰基磷脂酰乙醇胺-磷脂酶D的活性以及瞬时受体电位香草酸受体亚基1的结合未受影响。我们还发现，分别合成和降解2-花生四烯酰甘油的二酰基甘油脂肪酶和单酰基甘油脂肪酶不受FSH调节，这种内源性大麻素的膜转运也不受FSH调节。此外，我们发现蛋白激酶A（PKA）和细胞色素P450芳香化酶抑制剂可消除FSH对FAAH的刺激作用，相反，N,O'-二丁酰环磷腺苷和雌激素可模拟这种刺激作用。最后，我们证明FSH通过PKA和芳香化酶依赖性激活FAAH来保护支持细胞免受AEA的促凋亡活性影响。总之，这些数据表明，在整个内源性大麻素系统的各成分中，FAAH是FSH的唯一靶点，其通过PKA和芳香化酶依赖性途径的调节影响支持细胞的增殖。
