抗凋亡剂在人卵巢组织新型玻璃化冷冻法中的应用研究

Effect of Anti-apoptosis Agent on Human Ovarian Tissue Novel Vitrification

目的探讨在新型玻璃化冷冻法中添加抗凋亡剂是否改善人卵巢组织冷冻保存效果。方法收集2012年10月-2013年8月行妇科手术的患者卵巢标本共10例,使用新型玻璃化冷冻法保存。实验组冷冻保护液中添加抗凋亡剂维生素C,与未添加维生素C组(对照组)比较始基卵泡中卵母细胞和颗粒细胞超微结构、始基卵泡及间质凋亡发生率。结果实验组中卵母细胞和颗粒细胞超微结构正常百分率高于对照组(P<0.05)。在凋亡发生率检测中,实验组和对照组始基卵泡凋亡发生率分别为21.6%(49/227)、38.6%(91/236),差异有统计学意义(P<0.001)。卵巢间质凋亡分析中,实验组和对照组凋亡发生率分别为(21.33±3.41)%、(33.46±3.09)%,差异有统计学意义(P<0.001)。结论在新型玻璃化冷冻法中添加抗凋亡剂能更好地保存始基卵泡及周围间质细胞,改善玻璃化冷冻效果。

Objective To determine if the anti-apoptosis agent can improve the protective effect of human ovarian tissue with novel vitrification method. Methods Human ovarian cortical tissues were collected from ten patients treated between October 2012 and August 2013. The samples obtained from each patient were divided into two groups： control （the novel immersed vitrification） and experimental group （the novel immersed vitrification supplemented with vitamin C）. The preservation effects in the two groups were compared by ultrastructure using electronic microscopy, and apoptosis was assessed by terminal deoxynucleotidy1 transferase-medicated dUTP nick-end labeling （TUNEL） staining. Results The proportion of normal ultrastructure of oocytes and granulosa cells in the experimental group was higher than that in the control group （P 〈 0.05）. The proportion of TUNEL-positive primordial follicles in the experimental group was 21.6% （49/227） and the proportion of TUNEL-positive primordial follicles in the control group was 38.6% （91/236）, with a significant difference between the two groups （P 〈 0.001）. The proportion of TUNEL-positive stromal cells in the experimental group was （21.33±3.41） % and the proportion of TUNEL-positive stromal cells in the control group was （33.46±3.09） %, also with a significant difference between the two groups （P 〈 0.001）. Conclusions Anti-apoptosis agents can improve the preservation of primordial follicles and stromal cells by inhibiting of apoptosis. It may be a method worthy of trying in order to improve the protective effect of human ovarian tissue vitrification.