摘要
Infertility in females is the inability to become and maintain pregnancy, which can be caused by the abnormal development of a corpus luteum (CL). Abnormal development is linked to incomplete angiogenesis, which leads to an overall decrease in progesterone production. Angiogenesis is regulated by factors, such as fibroblast growth factor-2 (FGF-2), vascular endothelial growth factor (VEGF), and angiopoietin-1 (Ang-1) all of which are expressed in developing luteal tissue. Leptin, a potent satiety hormone, influences the expression of FGF and VEGF in non-ovarian tissue, and is also produced in the CL. Hence, it is hypothesized that leptin influences the production of FGF-2, VEGF, and Ang-1 in the developing CL. The objectives of the study were to (1) characterize angiogenic factor expression in the developing porcine CL and (2) determine the effect of leptin on FGF-2, VEGF, and Ang-1 expression in dispersed luteal tissue. Thirty mature crossbred gilts of similar age were allocated to one of five treatments (days of CL development: d 3, 4, 5, 6, or 7 of the estrous cycle; n=6/day) for tissue collection. Gilts were checked twice daily for classical, behavioral estrus using a boar for detection. Blood samples were collected on the day of CL harvest for serum progesterone analysis. Luteal tissue was divided and either embedded for immunohistochemistry, frozen in liquid nitrogen, or enzymatically digested and dispersed for culture. Dispersed cells were cultured with or without leptin (0, 10-12, 10 -11, 10-10, 10-9, 10-8 M; n = 3 wells/dose/gilt) for 24 hrs. Total expression for VEGF, FGF-2, Ang-1, and the leptin receptor (OB-Rb) was determined in all samples. All angiogenic factors and OB-Rb were localized to vascular endothelial cells, small luteal cells and large luteal cells; however, concentration and predominant cellular location varied by day of development. The expression of FGF-2 was highest (P = 0.05) on day 6 and leptin increased linearly (P = 0.005) as the CL developed. Leptin dose dependently decreased (P ≤ 0.05) VEGF, FGF-2 and Ang-1, relative to day of luteal development, by 10, 17 and 16.8%, respectively. Therefore, leptin appears to be involved in the angiogenic process in developing CL tissue.
摘要译文
女性不孕不能成为和维持妊娠，这可能是由黄体异常发展引起的（CL）。异常发展与不完全血管发生有关，导致孕酮生产总体下降。血管发生受因素调节，例如成纤维细胞生长因子-2（FGF-2），血管内皮生长因子（VEGF）和血管生成素-1（Ang-1），所有这些都在显影黄体组织中表达。瘦素，有效的饱腹感激素影响非卵巢组织中FGF和VEGF的表达，也在CL中产生。因此，假设瘦素影响FGF-2的产生，VEGF和Ang-1。研究目的是（1）鉴定发育中的猪CL中血管生成因子的表达，（2）确定瘦素对FGF-2的影响，VEGF和Ang-1在分散的黄体组织中的表达。将30只类似年龄的成熟杂交后备母猪分配到五种处理之一（CL发育的日期：d 3，4，5，6，或7发情周期; n \x3d 6 /天）用于组织收集。使用公猪进行检测，每天检查两次吉他，用于经典的行为发情。在CL收获当天收集血样用于血清孕酮分析。黄体组织分为免疫组织化学，液氮冷冻，或酶消化并分散培养。分散的细胞与或不与瘦素（0,10，-12，10 -11，10，10，10，-9， / sup，10 sup -8 / sup M;n \x3d 3孔/剂量/镀金）24小时。在所有样品中测定VEGF，FGF-2，Ang-1和瘦蛋白受体（OB-Rb）的总表达。所有血管生成因子和OB-Rb均定位于血管内皮细胞，小黄体细胞和大黄体细胞;然而，浓度和主要细胞位置随着发育日期而变化。在第6天，FGF-2的表达最高（P \x3d 0.05），CL发展时瘦素增加呈线性增加（P \x3d 0.005）。相对于黄体发育日，瘦素剂量依次降低（P≤0.05），VEGF，FGF-2和Ang-1分别为10,17和16.8％。因此，瘦素似乎参与血管生成过程中的发展中的CL组织。