Prevention of thecal angiogenesis, antral follicular growth, and ovulation in the primate by treatment with vascular endothelial growth factor Trap R1R2.
Prevention of thecal angiogenesis, antral follicular growth, and ovulation in the primate by treatment with vascular endothelial growth factor Trap R1R2.
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DOI：
10.1210/endo.143.7.8886
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发表时间：
2002-07
期刊：
影响因子：
4.8
通讯作者：
C. Wulff;H. Wilson;S. Wiegand;J. Rudge;H. Fraser
中科院分区：
文献类型：
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作者：
C. Wulff;H. Wilson;S. Wiegand;J. Rudge;H. Fraser
本研究旨在探讨抑制卵泡膜血管生成对绒猴卵泡发育的影响。为了抑制血管内皮生长因子（VEGF），在周期的卵泡期期间施用可溶性组合截短形式的fms样酪氨酸激酶（Flt）和与IgG融合的含有激酶插入结构域的受体（KDR）（VEGF陷阱R1 R2）10天。使用溴脱氧尿苷的免疫细胞化学来定量血管生成和滤泡细胞增殖的变化，以获得增殖指数，使用CD 31来可视化内皮细胞面积，并使用双重染色来区分膜内皮细胞增殖。通过测量卵泡直径、卵泡膜厚度和颗粒细胞增殖指数，通过形态学分析评估治疗对卵泡发育的影响。使用末端脱氧核苷酸转移酶-UTP缺口末端标记法检测和定量卵泡闭锁。用原位杂交技术研究VEGF及其受体Flt和KDR基因表达的影响。VEGF Trap R1 R2处理导致膜增殖和内皮细胞面积的显著减少，表明膜血管生成的抑制。缺乏正常的膜血管系统与膜厚度显著降低相关。窦卵泡发育严重受损，表现为颗粒细胞增殖减少、卵泡直径减小和排卵卵泡发育不足。此外，闭锁率显著增加。VEGF在颗粒细胞和卵泡膜细胞中的表达增加，而Flt和KDR在卵泡膜内皮细胞中的表达显著降低。这些结果表明，VEGF Trap治疗与卵泡血管生成的抑制有关，这导致窦卵泡发育和排卵的抑制。
This study was designed to investigate the effects of inhibition of thecal angiogenesis on follicular development in the marmoset monkey (Callithrix jacchus). To inhibit vascular endothelial growth factor (VEGF), a soluble combined truncated form of the fms-like tyrosine kinase (Flt) and kinase insert domain-containing receptor (KDR) receptor fused to IgG (VEGF Trap R1R2) was administered for 10 d during the follicular phase of the cycle. Changes in angiogenesis and follicular cell proliferation were quantified using immunocytochemistry for bromodeoxyuridine to obtain a proliferation index, CD31 to visualize endothelial cell area, and dual staining to distinguish thecal endothelial cell proliferation. The effects of the treatment on follicular development were assessed by morphometric analyses by measuring follicle diameter, thecal thickness, and a proliferation index for granulosa cells. Follicular atresia was detected and quantified using the terminal deoxynucleotidyltransferase-UTP nick end labeling method. Effects on gene expression of VEGF and its receptors, Flt and KDR, were studied by in situ hybridization. VEGF Trap R1R2 treatment resulted in a significant decrease in thecal proliferation and endothelial cell area, demonstrating the suppression of thecal angiogenesis. The absence of a normal thecal vasculature was associated with a significantly reduced thecal thickness. Antral follicular development was severely compromised, as indicated by decreased granulosa cell proliferation, decreased follicular diameter, and lack of development of ovulatory follicles. Furthermore, the rate of atresia was significantly increased. VEGF expression in granulosa and thecal cells increased after treatment, whereas Flt and KDR expressions in thecal endothelial cells were markedly decreased. These results show that VEGF Trap treatment is associated with the suppression of follicular angiogenesis, which results in the inhibition of antral follicular development and ovulation.