纳米顺铂对小鼠卵母细胞体外成熟的影响

Effect of Cisplatin Nanoparticles on in Vitro Maturation of Oocytes in Mice

纳米顺铂通过纳米载体包载顺铂进而发挥抗肿瘤作用,但纳米顺铂在杀死肿瘤细胞的同时,其对动物生殖细胞的影响尚不明确,试验通过在卵母细胞成熟培养液中添加顺铂及纳米顺铂对比了二者对小鼠卵母细胞体外成熟的影响。采集ICR小鼠未成熟卵母细胞并随机分为对照组、顺铂组和纳米顺铂组,体外培养16 h后统计卵母细胞成熟率;并对比卵丘卵母细胞复合体和裸卵经顺铂和纳米顺铂处理后减数分裂进程的差异,以探究颗粒细胞对纳米顺铂影响卵母细胞减数分裂的作用。通过免疫荧光染色检测染色体、纺锤体及肌动蛋白帽的形态变化以确定纳米顺铂对卵母细胞体外成熟过程中细胞骨架的影响。结果显示,顺铂组卵母细胞成熟率显著低于对照组,纳米顺铂组卵母细胞成熟率显著高于高浓度顺铂组,去除颗粒细胞后,纳米顺铂组停滞于GV期卵母细胞比例显著低于高浓度顺铂组。与对照组相比,顺铂组和纳米顺铂组的染色体排列正常率和细胞骨架形态正常率均显著降低,但纳米顺铂组染色体排列正常率显著高于顺铂组,两组之间细胞骨架正常率无显著性差异。结果提示,纳米顺铂通过增加减数分裂的完成性及维持核染色体的正常排布来减弱对小鼠卵母细胞的毒性作用,但不能改善顺铂对卵母细胞骨架的损伤,颗粒细胞能够在一定程度上降低纳米顺铂对卵母细胞减数分裂恢复的抑制作用。

Cisplatin nanoparticles exert anti-tumor effects by encapsulating cisplatin into nano-carriers,but its impact on animal germ cells remains unclear.This study compared the effect of cisplatin nanoparticles and cisplatin on the in vitro maturation of mouse oocytes by adding them to the oocyte culture medium.Immature oocytes of ICR mice were collected and randomly divided into control group,cisplatin group and cisplatin nanoparticles group.After 16 hours of in vitro culture,the maturation rate of oocytes was counted,differences in meiotic progression between cumulus-oocyte complexes and denuded oocytes treated with cisplatin and nanoparticle cisplatin were assessed to explore the role of granulosa cells in mediating the effects of nanoparticle cisplatin on oocyte meiosis.Immunofluorescent staining was used to detect morphological changes in chromosomes,spindles,and actin caps to determine the impact of cisplatin nanoparticle on the cytoskeleton during oocyte maturation in vitro.These results showed the maturation rate of oocytes in the cisplatin group was significantly lower than that in the control group,while the maturation rate of oocytes in the cisplatin nanoparticles group was significantly higher than that in the cisplatin group with high-concentration.After the removal of granule cells,the proportion of GV-stage oocytes in the cisplatin nanoparticles group was significantly decreased than that in the cisplatin group with high-concentration.Both the cisplatin group and the cisplatin nanoparticles group had a significantly lower rate of normal chromosomes alignment and normal cytoskeleton morphology compared with the control group.However,the rate of normal chromosome arrangement in the cisplatin nanoparticles group was significantly higher than that in the cisplatin group,and there was no significant difference in the rate of normal cytoskeleton between the two groups.These findings suggested that cisplatin nanoparticles can weaken the toxic effect of cisplatin on mouse oocytes by enhancing the completion of meiosis and maintaining the normal distribution of chromosome,but cannot reduce the damage of cisplatin to the cytoskeleton,granulosa cells can partly reduce the inhibition of cisplatin nanoparticles on oocyte meiosis recovery.