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采用nano LC-LTQ-Orbitrap-MS/MS蛋白质组学方法筛选鉴定香附四物汤干预原发性痛经模型小鼠卵巢组织中的差异表达蛋白,推测香附四物汤治疗原发性痛经的机制。采用雌激素和缩宫素联合造成原发性痛经模型,给药组连续灌胃给予香附四物汤3 d,于末次给药后1 h,取卵巢,经蛋白提取、变性、还原、烷基化、脱盐、酶解后,进行nano LC分离并在线连接电喷雾串联Orbitrap质谱分析,利用Thermo Proteome discoverer 1.4进行数据处理,筛选并鉴定差异表达蛋白,对差异表达蛋白进行生物信息学分析。随后采用Western blot法,将重要差异蛋白进行验证。正常组、模型组、给药组3组共有的差异蛋白106个,参与细胞过程的蛋白所占比例较大,其所参与的黏着连接、黏着斑等信号通路对细胞内各种信号通路均有调节作用。验证的ADRM1蛋白在模型组中,较正常组表达上调;给药后,较模型组表达下调。通过有效的比较分析,得到一些香附四物汤干预原发性痛经的差异表达蛋白,其中ADRM1蛋白有可能是香附四物汤作用的靶点。
Screening and Identification of Differentially Expressed Proteins in the Mouse Model of Primary Dysmenorrhea by Using LC-LTQ-Orbitrap-MS / MS Proteomics Method mechanism. The model of primary dysmenorrhea was induced by combination of estrogen and oxytocin. The medicine group was given gavage for 3 days continuously, and ovariectomized 1 h after the last administration. The ovariectomized mice were subjected to protein extraction, denaturation, reduction, After the hydrolysis, hydrolysis and enzymolysis, nano LC separation and tandem electrospray tandem Orbitrap mass spectrometry were carried out, and the data were processed by Thermo Proteome discoverer 1.4. The differentially expressed proteins were screened and identified, and the differential expressed proteins were analyzed by bioinformatics. Subsequently, Western blot was used to validate important differential proteins. There were 106 differentially expressed proteins in the normal group, the model group and the drug-administered group, and the protein involved in cellular processes accounted for a large proportion. The involved signaling pathways such as adhesion junction and focal adhesions had a significant effect on various intracellular signaling pathways Regulatory effect. The validated ADRM1 protein was upregulated in the model group compared with the normal group; after administration, the expression of ADRM1 was down-regulated compared with the model group. Through effective comparative analysis, we obtained some differentially expressed proteins of Xiang-Fu-Si-Wu Decoction for primary dysmenorrhoea. Among them, ADRM1 protein may be the target of the effect of Xiangfu Siwu Decoction.