摘要
目的研究人类白细胞抗原G（human leukocyte antigen，HLA-G）及其异构体mRNA在人类着床前胚胎的表达，探讨HLA-G在人类胚胎早期发育中的作用和意义。方法第四军医大学唐都医院妇产科生殖医学中心2003-01-2005-12期间，以体外受精-胚胎移植技术助孕过程中剩余的患者自愿捐赠的人类早期胚胎作为研究对象，采用巢式RT-PCR检测早期胚胎HLA-G及其异构体mRNA的表达。结果检测20个受精后2—3d的人类卵裂期胚胎，2、4、6、8细胞期胚胎各5个，有7个胚胎表达HLA-G及其部分的异构体mRNA。检测5个受精后4d的人类桑葚胚，全部表达HLA-G及其部分的异构体mRNA。检测25个受精后6d的扩张期囊胚，全部受检囊胚均表达HLA-G mRNA，但异构体表达不同。在受检的25个扩张期囊胚中，20个表达HLA-G1（表达率80．0％），4个表达G2（表达率16．0％），25个表达G3（表达率100％）、24个表达G4（表达率96．0％），5个表达G5（表达率20．0％），8个表达G6（表达率32．0％）。结论着床前的人类胚胎表达HLA-G及其异构体mRNA，其阳性表达胚胎的比例，随着胚胎发育的进程而增加。HLA-G异构体在人类着床前胚胎差异表达。
Objective To investigate the expression of HLA-G （ human leukocyte antigen-G） and its isoforms in human preimplantation embryos in order to discuss the significance of HLA-G in the early embryo development. Methods The human embryos were donered by the volunteers from the residual fertilized ovum after accepting IVF from January of 2003 to December of 2005 in the Reproductive Center of Tangdu Hospital, Fourth Military Medical University. Human embryo mRNA was isolated using Dynabeads mRNA DIRECTI＇M Kit. Nested RT-PCR was performed using SuperScriptTM One-Step RT-PCR Platinum Taq Kit and Platinum PCR SuperMix High Fidelity Kit. Results HLA-G and its isoforms in human cleavage stage embryos：a total of twenty individual cleavage stage human embryos were examined （five 2-cell, five 4-cell, five 6-cell and five 8-cell）. Only 7/20 embryos expressed HLA-G and its partial isoforms mRNA. HLA-G mRNA isoforms in human morulas：five individual four-day human embryos were examined. All of them expressed HLA-G and its partial isoforms mRNA. HLA-G mRNA isoforms in human blastocysts：twenty-five six-day expanded blastocysts were studied. All of them were found to express HLA-G but different isoforms mRNA. G1 was detected in 20/25 （80%）, G2 in 4/35 （ 16% ）, G3 in 25/25 （ 100% ）, G4 in 24/25 （96%）, G5 in 5/25 （20%） and G6 in 8/25 （ 32% ）. Conclusions The HLA-G expression increases along with the stages of embryo development. Human preimplantation embryos diferentially express HLA-G mRNA isoforms in different development stages.
出处
《中国实用妇科与产科杂志》
CAS
CSCD
北大核心
2006年第7期505-508,共4页
Chinese Journal of Practical Gynecology and Obstetrics
基金
国家自然科学基金资助项目(30471812)
