二补助育改良方对反复体外受精-胚胎移植失败女性子宫内膜影响的蛋白组学研究

Proteomic study on the effects of modified Erbu Zhuyu Decoction on endometrium in womenwithrepeated IVF-ETfailure

目的:借助蛋白质谱法研究二补助育改良方对反复体外受精-胚胎移植(IVF-ET)失败女性患者子宫内膜容受性影响的蛋白组学差异。方法:收集反复IVF-ET失败,二补助育改良方药物干预3个月及未用药干预患者资料,采集两组患者(各3例)黄体中期子宫内膜组织,进行蛋白质提取、酶切后iTRAQ标记、液-质联用分析,获得质谱检测数据。利用R语言limma包进行蛋白差异分析(FDR<0.05,Ilog2FCl>1.2)、以及基因富集分析生物学功能(GO)、信号通路(KEGG)及蛋白相互作用(PPI)。结果:共发现有166个差异表达蛋白,其中上调蛋白111个、下调蛋白55个。生物学进程富集到基因转录、蛋白合成相关蛋白,细胞成分和生物学功能富集在核糖体、内质网以及细胞黏附相关蛋白。其中包括紧密连接蛋白1(TJP1)、基质金属蛋白酶2(MMP2)、同源盒基因A11(HOXA11)及层黏连蛋白2(LAMA2)等高表达,并以免疫组化进行验证,结果显示试验组相关指标高表达。结论:二补助育改良方对反复IVF-ET失败患者子宫内膜蛋白组学多种细胞因子具有调节作用,促进了细胞蛋白合成、细胞运动、黏附等,这可能是二补助育改良方改善反复IVF-ET失败患者子宫内膜容受性的效用机制。

Objective:To study the proteomic difference of modified Erbu Zhuyu Decoction on endometrial receptivity in women with repeated IVF-ET failure by protein mass spectrometry.Methods:Collected data of patients with repeated IVF-ET failure,who received three months of modified Erbu Zhuyu Decoction intervention and no drug intervention,collected mid-luteal endometrial tissue of two group patients(3 cases in each group),protein extraction,iTRAQ labeling after digestion,LC-MS analysis to obtain mass detection data,using R language software to analysis protein differences(FDR<0.05,Jlog2FC>1.2),biological function and other go analysis,Signal path analysis(KEGG)and protein interaction analysis.Results:A total of 166 differentially expressed proteins were found,including 111 up-regulated proteins and 55 down-regulated proteins.Biological processes were enriched with gene transcription and protein synthesis related protein,and cellular components and biological functions were enriched in ribosomes,endoplasmic reticulum,and cell adhesion-related proteins,including high expression of TJP1,MMP2,HOXA11 and LAMA2 proteins,which were verified by immunohistochemistry.Conclusion:There are significant differences in the expression of multiple cytokines in proteomics with or without modified Erbu Zhuyu Decoction intervention in patients with repeated IVF-ET failure,by promoting cell protein synthesis,cell movement,adhesion,etc.This may be the effective mechanism of modified Erbu Zhuyu Decoction to improve endometrial receptivity in patients with repeated IVF-ET failure.