Laboratory of Tumor and Developmental Biology, GIGA-R, University of Liège, Tour de Pathologie (B23), Avenue Hippocrate, 13, Sart Tilman, B-4000, Liège, Belgium.
Department of Gynecology and Obstetrics, CPMA, University of Liège, CHR de la Citadelle, Boulevard du 12ième de Ligne, 1, B-4000, Liège, Belgium.
J Assist Reprod Genet. 2019 Feb;36(2):349-359. doi: 10.1007/s10815-018-1353-8. Epub 2018 Nov 3.
To evaluate the efficiency of ovarian tissue treatment with Z-VAD-FMK, a broad-spectrum caspase inhibitor, to prevent follicle loss induced by ischemia/reperfusion injury after transplantation.
In vitro, granulosa cells were exposed to hypoxic conditions, reproducing early ischemia after ovarian tissue transplantation, and treated with Z-VAD-FMK (50 μM). In vivo, cryopreserved human ovarian fragments (n = 39) were embedded in a collagen matrix containing or not Z-VAD-FMK (50 μM) and xenotransplanted on SCID mice ovaries for 3 days or 3 weeks.
In vitro, Z-VAD-FMK maintained the metabolic activity of granulosa cells, reduced HGL5 cell death, and decreased PARP cleavage. In vivo, no improvement of follicular pool and global tissue preservation was observed with Z-VAD-FMK in ovarian tissue recovered 3-days post-grafting. Conversely, after 3 weeks of transplantation, the primary follicular density was higher in fragments treated with Z-VAD-FMK. This improvement was associated with a decreased percentage of apoptosis in the tissue.
In situ administration of Z-VAD-FMK slightly improves primary follicular preservation and reduces global apoptosis after 3 weeks of transplantation. Data presented herein will help to guide further researches towards a combined approach targeting multiple cell death pathways, angiogenesis stimulation, and follicular recruitment inhibition.
评估广谱半胱天冬酶抑制剂 Z-VAD-FMK 处理卵巢组织以预防移植后缺血/再灌注损伤引起的卵泡丢失的效率。
在体外，用缺氧条件（复制卵巢组织移植后早期的缺血）处理颗粒细胞，并使用 Z-VAD-FMK（50μM）进行处理。在体内，将冷冻保存的人类卵巢碎片（n=39）嵌入含有或不含有 Z-VAD-FMK（50μM）的胶原基质中，并在 SCID 小鼠卵巢上进行 3 天或 3 周的异种移植。
在体外，Z-VAD-FMK 维持了颗粒细胞的代谢活性，减少了 HGL5 细胞的死亡，并减少了 PARP 的切割。在体内，在移植后 3 天回收的卵巢组织中，Z-VAD-FMK 对卵泡池和整体组织保存没有改善。相反，在移植 3 周后，用 Z-VAD-FMK 处理的碎片中初级卵泡密度更高。这种改善与组织中凋亡细胞的百分比降低有关。
原位给予 Z-VAD-FMK 可轻微改善初级卵泡的保存，并减少移植后 3 周的整体凋亡。本文提供的数据将有助于指导进一步的研究，以针对多种细胞死亡途径、血管生成刺激和卵泡募集抑制的联合方法。
