Hernández-Coronado C G, Guzmán A, Espinosa-Cervantes R, Romano M C, Verde-Calvo J R, Rosales-Torres A M
1Departamento Producción Agrícola y Animal,Universidad Autónoma Metropolitana-Xochimilco,Calzada del Hueso 1100,CP 04960,México D.F.,México.
2CINVESTAV,I.P.N. Departamento de Fisiología,Biofísica y Neurociencias,Av. Instituto Politécnico Nacional 2508,Código Postal 07360,México D.F.,México.
Animal. 2015 Feb;9(2):308-12. doi: 10.1017/S1751731114002341. Epub 2014 Sep 23.
The follicle destiny towards ovulation or atresia is multi-factorial in nature and involves outcries, paracrine and endocrine factors that promote cell proliferation and survival (development) or unchain apoptosis as part of the atresia process. In several types of cells, sphingosine-1-phospate (S1P) promotes cellular proliferation and survival, whereas ceramide (CER) triggers cell death, and the S1P/CER ratio may determine the fate of the cell. The aim of present study was to quantify S1P and CER concentrations and their ratio in bovine antral follicles of 8 to 17 mm classified as healthy and atretic antral follicles. Follicles were dissected from cow ovaries collected from a local abattoir. The theca cell layer, the granulosa cells and follicular fluid were separated, and 17β-estradiol (E2) and progesterone (P4) concentrations were measured in the follicular fluid by radioimmunoassay. Based on the E2/P4 ratio, the follicles were classified as healthy (2.2±0.3) or atretic (0.2±0.3). In both follicular compartments (granulosa and theca cell layer), sphingolipids were extracted and S1P and CER concentrations were quantified by HPLC (XTerra RP18; 5 µm, 3.0×150 mm column). Results showed that in both follicular compartments, S1P concentrations were higher in healthy antral follicles than in atretic antral follicles (P<0.05). The concentration of CER in the granulosa cells was higher in atretic antral follicles than in healthy antral follicles, but no differences were observed in the theca cell layer. The S1P/CER ratio in both follicular compartments was also higher in healthy antral follicles. Interestingly, in these follicles, there was a 45-fold greater concentration of S1P than CER in the granulosa cells (P<0.05), whereas in the theca cell layer, S1P had only a 14-fold greater concentration than CER when compared with atretic antral follicles. These results suggest that S1P plays a role in follicle health, increasing cellular proliferation and survival. In contrast, reduction of S1P and the S1P/CER in the antral follicle could trigger cellular death and atresia.
卵泡走向排卵或闭锁的命运本质上是多因素的，涉及促进细胞增殖和存活（发育）的自分泌、旁分泌和内分泌因素，或引发作为闭锁过程一部分的细胞凋亡。在几种类型的细胞中，鞘氨醇-1-磷酸（S1P）促进细胞增殖和存活，而神经酰胺（CER）触发细胞死亡，S1P/CER 比值可能决定细胞的命运。本研究的目的是量化分类为健康和闭锁的 8 至 17 毫米牛窦状卵泡中 S1P 和 CER 的浓度及其比值。从当地屠宰场收集的母牛卵巢中解剖卵泡。分离出卵泡膜细胞层、颗粒细胞和卵泡液，并通过放射免疫测定法测量卵泡液中 17β-雌二醇（E2）和孕酮（P4）的浓度。根据 E2/P4 比值，将卵泡分类为健康（2.2±0.3）或闭锁（0.2±0.3）。在两个卵泡区室（颗粒细胞和卵泡膜细胞层）中，提取鞘脂并通过高效液相色谱法（XTerra RP18；5 µm，3.0×150 mm 柱）定量 S1P 和 CER 的浓度。结果表明，在两个卵泡区室中，健康窦状卵泡中的 S1P 浓度均高于闭锁窦状卵泡（P<0.05）。闭锁窦状卵泡中颗粒细胞的 CER 浓度高于健康窦状卵泡，但在卵泡膜细胞层未观察到差异。两个卵泡区室中健康窦状卵泡的 S1P/CER 比值也更高。有趣的是，在这些卵泡中，颗粒细胞中 S1P 的浓度比 CER 高 45 倍（P<0.05），而在卵泡膜细胞层中，与闭锁窦状卵泡相比，S1P 的浓度仅比 CER 高 14 倍。这些结果表明，S1P 在卵泡健康中起作用，增加细胞增殖和存活。相反，窦状卵泡中 S1P 和 S1P/CER 的减少可能触发细胞死亡和闭锁。
