Grazul-Bilska A T, Redmer D A, Jablonka-Shariff A, Biondini M E, Reynolds L P
Department of Animal and Range Sciences, North Dakota State University, Fargo 58105, USA.
Can J Physiol Pharmacol. 1995 Apr;73(4):491-500. doi: 10.1139/y95-062.
This study was conducted to evaluate the effects of fibroblast growth factor 1 (FGF-1), fibroblast growth factor 2 (FGF-2), or luteinizing hormone (LH) on proliferation and progesterone secretion of ovine luteal cells from days 5, 10, or 15 after estrus (estrus = day 0; n = 4 or 5 ewes/day). After enzymatic dispersion, luteal cells were incubated in the presence or absence of various doses of FGF-1, FGF-2, LH, or fetal bovine serum (FBS) (positive control) in serum-free media for 7 days in 24-well plates. Cells were counted on day 7 of culture and media analyzed for progesterone concentration. For all treatments, maximal effects (Emax) and dissociation constants (KD) were calculated. In addition, luteal cells were cultured in eight-chamber slides and treated as above, but on day 7 of culture cells were fixed and stained for the presence of 3beta-hydroxy-delta 5-steroid dehydrogenase (3beta HSD). The number of steroidogenic (3beta HSD positive) cells per unit area was counted for control cultures (no treatment) and cultures treated with the most effective doses of FGF-1, FGF-2, LH, OR FBS in proliferation and (or) progesterone assays. FGF-1, FGF-2, AND FBS stimulated (p < 0.05) proliferation of luteal cells from all stages of luteal development in a dose-dependent manner. In addition, LH increased (p < 0.01) the number of 3beta HSD-positive cells across all stages of luteal development. Moreover, LH and FBS increased (p < 0.05) progesterone secretion by luteal cells from all stages in a dose-responsive manner, but the effects of FGF-1 and FGF-2 were variable. For proliferation, the Emax of all factors was greatest (p < 0.01) on day 5, whereas the KD values were similar across days of the estrous cycle. For progesterone production, the Emax and KD of LH and FBS were similar and did not differ across the estrous cycle. These data demonstrate the luteal cells from the early luteal phase of the estrous cycle exhibit the greatest ability to proliferate and (or) increase their progesterone secretion in response to FGF-1, FGF-2, LH, or FBS. In addition, although LH does not affect the total number of luteal cells in culture, it does increase the number of steroidogenic cells. These data indicate that in addition to LH, fibroblast growth factors may be involved in regulation of luteal growth and differentiation in ewes.
本研究旨在评估成纤维细胞生长因子1（FGF-1）、成纤维细胞生长因子2（FGF-2）或促黄体生成素（LH）对发情后第5、10或15天绵羊黄体细胞增殖和孕酮分泌的影响（发情日=第0天；每组n = 4或5只母羊/天）。酶解分散后，将黄体细胞在无血清培养基中，于24孔板中在有或无不同剂量的FGF-1、FGF-2、LH或胎牛血清（FBS）（阳性对照）存在的情况下培养7天。在培养第7天对细胞进行计数，并分析培养基中的孕酮浓度。对于所有处理，计算最大效应（Emax）和解离常数（KD）。此外，将黄体细胞培养在八孔载玻片上并按上述方法处理，但在培养第7天固定细胞并染色以检测3β-羟基-Δ5-类固醇脱氢酶（3β HSD）的存在。对对照培养物（未处理）以及在增殖和（或）孕酮测定中用最有效剂量的FGF-1、FGF-2、LH或FBS处理的培养物，计数每单位面积的类固醇生成（3β HSD阳性）细胞数量。FGF-1、FGF-2和FBS以剂量依赖性方式刺激（p < 0.05）黄体发育各阶段黄体细胞的增殖。此外，LH增加（p < 0.01）黄体发育各阶段3β HSD阳性细胞的数量。而且，LH和FBS以剂量反应性方式增加（p < 0.05）各阶段黄体细胞的孕酮分泌，但FGF-1和FGF-2的作用存在差异。对于增殖，所有因子的Emax在第5天最大（p < 0.01），而KD值在发情周期各天相似。对于孕酮产生，LH和FBS的Emax和KD相似，且在发情周期各天无差异。这些数据表明，发情周期黄体早期的黄体细胞对FGF-1、FGF-2、LH或FBS的反应表现出最大的增殖能力和（或）增加其孕酮分泌的能力。此外，尽管LH不影响培养中黄体细胞的总数，但它确实增加了类固醇生成细胞的数量。这些数据表明，除LH外，成纤维细胞生长因子可能参与母羊黄体生长和分化的调节。
