Kim Yong-Soo, Kim Myung-Sun, Lee Sook-Hwan, Choi Bum-Chae, Lim Jeong-Mook, Cha Kwang Yul, Baek Kwang-Hyun
Cell and Gene Therapy Research Institute, Pochon CHA University, CHA General Hospital, Seoul, Korea.
Proteomics. 2006 Jun;6(11):3445-54. doi: 10.1002/pmic.200500775.
Recurrent spontaneous abortion (RSA), defined as the loss of three or more consecutive pregnancies prior to the 20th week of gestation, affects up to 5% of the child-bearing population. To investigate the proteins associated with RSA, the protein expression in human follicular fluid was analyzed using 2-DE. Follicular fluid contains a variety of biologically important proteins for oocyte fertilization and follicle maturation in the mammalian reproductive process. Therefore, it can be used as a provisional source for identifying proteins involved in RSA. In this study, we identified five aberrantly expressed proteins (complement component C3c chain E, fibrinogen gamma, antithrombin, angiotensinogen, and hemopexin precursor) in follicular fluid from RSA patients with MALDI-TOF-MS and nano-LC MS/MS. Western blot analysis confirmed that the protein expression level of fibrinogen gamma and antithrombin was less in follicular fluid from RSA patients than those from normal controls. Semiquantitative RT-PCR and real-time PCR analyses revealed that mRNA level of these coagulation factors was also decreased significantly in chorionic villi of RSA patients compared with normal samples. Taken all together, it is likely that coagulation factors (fibrinogen gamma and antithrombin) play an important role in maintaining the normal pregnancy.
复发性自然流产（RSA）被定义为妊娠20周前连续发生三次或更多次妊娠丢失，影响着高达5%的育龄人群。为了研究与RSA相关的蛋白质，使用双向电泳（2-DE）分析了人卵泡液中的蛋白质表达。卵泡液在哺乳动物生殖过程中含有多种对卵母细胞受精和卵泡成熟具有生物学重要性的蛋白质。因此，它可作为鉴定参与RSA的蛋白质的临时来源。在本研究中，我们通过基质辅助激光解吸电离飞行时间质谱（MALDI-TOF-MS）和纳升液相色谱串联质谱（nano-LC MS/MS）鉴定了RSA患者卵泡液中五种异常表达的蛋白质（补体成分C3c链E、纤维蛋白原γ、抗凝血酶、血管紧张素原和血红素结合蛋白前体）。蛋白质印迹分析证实，RSA患者卵泡液中纤维蛋白原γ和抗凝血酶的蛋白质表达水平低于正常对照者。半定量逆转录聚合酶链反应（RT-PCR）和实时PCR分析显示，与正常样本相比，RSA患者绒毛膜中这些凝血因子的mRNA水平也显著降低。综上所述，凝血因子（纤维蛋白原γ和抗凝血酶）可能在维持正常妊娠中起重要作用。
