补肾活血方调节在位内膜蛋白质表达治疗子宫内膜异位症不孕机制研究

Mechanism of Regulating Ectopic Endometrium Protein Expression by Bushen Huoxue Recipe in Treatment of Endometriosis Associated Infertility

目的:比较观察子宫内膜异位症(endometriosis,Ems)相关不孕患者与正常育龄期妇女在位子宫内膜的蛋白质表达差异及中药补肾活血方对其调节作用。方法:提取正常育龄期妇女、EMs不孕患者服用补肾活血方治疗3个月前后的分泌中期子宫内膜各4例,利用差速离心和密度梯度离心方法来提取纯化内膜蛋白质,采用荧光差异凝胶电泳(2D-DIGE)标记、分离蛋白质,Decyder差异分析软件进行分析、识别差异表达蛋白质,并应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF/TOF)得到相应差异蛋白的肽质量指纹图谱,然后搜索数据库鉴定部分差异蛋白质点。结果:分析得到35个差异蛋白点,质谱鉴定出28种差异蛋白。结论:分析预测Ems不孕发生的5种可能机制,涉及10种蛋白;补肾活血方作用于26种蛋白从4种可能机制修复Ems不孕患者在位内膜缺陷。

Objective： To compare the uterus protein expression differences between endometriosis （Ems） correlation infertile patients and normal women of reproductive age, and study the mechanism of Bushen Huoxue recipe （BSHXR）. Method. Four cases of Ems infertility patients with kidney deficiency and blood stasis syndrome diagnosed by laparoscopic surgery wereobserved. Ectopic endometria were collected during the laparoscopic surgery as Ems group. BSHXR was given to Eros patients after the surgery, lasting for three months, the ectopic endometria were collected after the treatment course as BSHXR group. Besides, four healthy women＇s endometria were collected three months after drug-induced abortion as normal control group. All the endometrial samples were pathologically diagnosed as secretory phase endometrium. Differential centrifugation and density gradient centrifugation method were applied to extract purified endometrial proteins. Difference gel electrophoresis （DIGE） technology were used to get the expression patterns of ectopic endometria differential proteome. Decyder difference 2-D analysis software were used to analysis. Matrix-assisted laser desorption ionization mass spectrometry （MALDI-TOF/TOF） were used to identify among these differential protein spots. Result： Through establishing the differential proteome technology platform of ectopic endometrial proteins among 3 groups. 35 different express proteins were found by differential software analysis, 28 different proteins were identified by using mass spectrometry identification. Conclusion： Five possible mechanisms of Eros-associated infertility were predicted involving 10 different express proteins. BSHXR can regulate 26 different express proteins by 4 possible mechanisms to treat ectopic endometria of patients with Ems-associated infertility.