摘要
目的建立彗星分析小鼠卵母细胞凋亡的模型,研究靶向酸性鞘磷酯酶1(SMPD1)的siRNA对卵子自发性凋亡的保护作用,为建立新的生殖保护方法奠定基础。方法通过同源性分析并结合siRNA设计软件设计并化学合成靶向SMPD1的3条siRNA,用超排卵技术对雌性BALB/c小鼠进行超排卵并杀鼠取卵,利用显微注射的方法将siRNA分别导入超排获得的小鼠卵子,分别于48h和72h观察卵子形态学变化,通过彗星分析检测卵细胞DNA破坏程度来分析卵子自发性凋亡情况。结果彗星分析法观察到小鼠卵母细胞在体外培养24h可见少量DNA泳出,48h可见大量DNA泳出;通过显微注射导入siRNA48h后的彗星分析结果显示,其中1对siRNA注射组,即siRNA003组的卵母细胞泳出量与其它2个siRNA注射组及对照组相比显著降低(P<0.01),其它2对siRNA注射组与对照组无明显差异。结论靶向SMPD1的siRNA能够有效保护卵子的自发性凋亡,有望成为新的生殖保护方法。
Objective To investigate the potential of siRNAs targeting sphingomyelin phosphodiesterase 1（SMPD1） in protecting the oocytes from apoptosis,and explore new approaches to female fertility preservation.Methods Chemically synthesized siRNA targeting SMPD1 were introduced into mouse oocytes retrieved by hyperstimulation,and the cell apoptosis was analyzed by comic assay 48 and 72 h later.Results In the occytes without any siRNA injection,occyte DNA damage occurred after 24 h,and large amount of DNA fragments migrated from the cells 48 h later.In occytes injected with siRNA003,DNA migration decreased significantly as compared with the control and the other two groups injected with siRNA001 and siRNA002（P〈0.01）.Conclusion siRNA targeting SMPD1 may protect the oocytes from apoptosis,and has the potential for use in future female fertility preservation.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2009年第11期2165-2167,2170,共4页
Journal of Southern Medical University
基金
国家自然科学基金(30400474
30700327)
广东省自然科学基金(4009374
9151009101000011
7300047)
广东省医学科研基金(B2004038
B2006003)
广东省人口计生委科研项目(2009227)
