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远程缺血预处理下新鲜大鼠卵巢自体移植中细胞凋亡、增殖和血管生成的进展性评估

Progressive Evaluation of Apoptosis, Proliferation, and Angiogenesis in Fresh Rat Ovarian Autografts Under Remote Ischemic Preconditioning.

作者信息

Damous Luciana Lamarão, Silva Sônia Maria da, Carbonel Adriana Aparecida Ferraz, Simões Manuel de Jesus, Baracat Edmund Chada, Montero Edna Frasson de Souza

机构信息

Laboratory of Structural and Molecular Gynecology (LIM-58), Departamento de Ginecologia e Obstetrícia, Faculdade de Medicina da Universidade de São Paulo (FMUSP), São Paulo, Brasil

Universidade da Cidade de São Paulo (Unicid), São Paulo, Brasil.

出版信息

Reprod Sci. 2016 Jun;23(6):803-11. doi: 10.1177/1933719115620493. Epub 2015 Dec 16.

Abstract

This study evaluated the remote ischemic preconditioning (R-IPC) early and late repercussion on fresh ovarian transplants, aiming to assess a probable protective effect in ovarian follicular pool. Sixty Wistar EPM-1 rats were used, divided in 2 study groups: ovarian transplantation (Tx) and Tx + R-IPC, submitted to ovary transplant with or without R-IPC, respectively. These groups were subdivided according to the date for euthanasia: 4th, 7th, 14th, 21st, and 30th days of the postoperatory period. Morphology, morphometry, neoangiogenesis (vascular endothelial growth factor [VEGF]), proliferative activity (Ki-67), and apoptosis (cleaved caspase-3) were evaluated. Remote ischemic preconditioning was performed in the common iliac artery. Fresh autologous ovarian tissue was implanted integrally in the retroperitoneum. All animals showed resumption of estrous phase after ovary transplantation. Remote ischemic preconditioning attenuated the lesions progressively from the 7th day, with greater number of the immature follicles (14 days, P < .05), but didn't affect mature follicles and corpora lutea (P > .05). Immunohistochemical analyzes, taken as a whole, show that R-IPC benefic effect is more evident in the later periods of evaluation, when a greater proliferative activity (14, 21, and 30 days, P < .05) and lesser cell apoptotic activity (21 and 30 days, P < .05). The VEGF expression was similar in all times (P > .05). Remote ischemic preconditioning could have a benefic effect in the progressive evaluation of freshly grafted ovarian, especially on the latest phases of the posttransplant period. The 14th day was a landmark in the recuperation of the graft. Further investigations are necessary to determine the role of R-IPC in this scenario and its effect in frozen-thawed ovarian tissue.

摘要

本研究评估了远程缺血预处理(R-IPC)对新鲜卵巢移植的早期和晚期影响,旨在评估其对卵巢卵泡池可能的保护作用。使用了60只Wistar EPM-1大鼠,分为2个研究组:卵巢移植(Tx)组和Tx + R-IPC组,分别接受有或无R-IPC的卵巢移植。这些组根据安乐死日期进一步细分:术后第4、7、14、21和30天。评估了形态学、形态计量学、新生血管形成(血管内皮生长因子[VEGF])、增殖活性(Ki-67)和凋亡(裂解的半胱天冬酶-3)。在髂总动脉进行远程缺血预处理。将新鲜的自体卵巢组织完整植入腹膜后。所有动物在卵巢移植后均恢复了发情期。远程缺血预处理从第7天开始逐渐减轻损伤,未成熟卵泡数量增加(第14天,P < 0.05),但不影响成熟卵泡和黄体(P > 0.05)。总体而言,免疫组织化学分析表明,R-IPC的有益作用在评估后期更为明显,此时增殖活性更高(第14、21和30天,P < 0.05),细胞凋亡活性更低(第21和30天,P < 0.05)。VEGF表达在所有时间均相似(P > 0.05)。远程缺血预处理可能对新鲜移植卵巢的渐进性评估有有益作用,尤其是在移植后的后期阶段。第14天是移植物恢复的一个里程碑。有必要进一步研究以确定R-IPC在这种情况下的作用及其对冻融卵巢组织的影响。

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