基于蛋白组学探究七味癸宝颗粒治疗卵巢早衰的作用机制

Mechanism of Qiwei Guibao Granules in treatment of premature ovarian failure based on proteomics

利用蛋白质组学技术研究七味癸宝颗粒治疗卵巢早衰(premature ovarian failure,POF)的可能机制。首先使用50 mg·kg-1剂量的雷公藤多苷溶液连续灌胃14 d,构建POF小鼠模型。造模结束的前10 d,每天观察小鼠的动情周期,以判断是否造模成功。然后,从造模结束后的第1天开始,用七味癸宝颗粒对POF模型小鼠每天进行灌胃给药,连续治疗4周。实验结束第2天,小鼠摘眼球取血,离心后分离出血清;取出小鼠卵巢和子宫,小心剥离脂肪组织,计算各组小鼠的卵巢和子宫的器官指数。ELISA检测各组小鼠血清中雌激素的水平。卵巢组织提取蛋白样品,运用串联质谱标签(tandem mass tags,TMT)定量蛋白质组学技术研究七味癸宝颗粒干预前后和造模前后的差异蛋白情况。差异蛋白分析结果显示,七味癸宝颗粒的治疗能够调节雷公藤多苷诱导的POF模型相关的26个差异表达蛋白,主要有S100A4、STAR、adrenodoxin oxidoreductase、XAF1、PBXIP1等。GO富集结果显示,26个差异蛋白主要富集在生物过程和细胞组分。KEGG富集结果显示,这些差异蛋白主要涉及complement and coagulation cascades、focal adhesion、arginine biosynthesis及terpenoid backbone biosynthesis等信号通路。其中complement and coagulation cascades可能是七味癸宝颗粒治疗POF的潜在靶通路。该研究利用蛋白质组学筛选出七味癸宝颗粒治疗雷公藤多苷所致小鼠POF的相关差异蛋白,主要参于免疫调节、凋亡调节、补体和凝血级联反应、胆固醇代谢与类固醇激素生成,这些可能是七味癸宝颗粒治疗POF的主要作用机制。

In this study,the underlying mechanism of Qiwei Guibao Granules(QWGB)in the treatment of premature ovarian fai-lure(POF)was explored by the proteomics technique.Firstly,the POF model was induced in mice by intragastric administration of Tripterygium wilfordii glycosides solution at 50 mg·kg-1for 14 days.Ten days prior to the end of the modeling,the estrous cycle of mice was observed every day to evaluate the success of modeling.From the 1st day after modeling,the POF model mice were treated with QWGB by gavage every day and the treatment lasted four weeks.On the 2nd day after the end of the experiment,blood was collected from the eyeballs and the serum was separated by centrifugation.The ovaries and uterus were collected and the adipose tissues were carefully stripped.The organ indexes of the ovaries and uterus of each group were calculated.The serum estrogen(E2)level of mice in each group was detected by ELISA.Protein samples were extracted from ovarian tissues of mice,and the differential proteins before and after QWGB intervention and before and after modeling were analyzed by quantitative proteomics using tandem mass tags(TMT).As revealed by the analysis of differential proteins,QWGB could regulate 26 differentially expressed proteins related to the POF model induced by T.wilfordii glycosides,including S100A4,STAR,adrenodoxin oxidoreductase,XAF1,and PBXIP1.GO enrichment results showed that the 26 differential proteins were mainly enriched in biological processes and cellular components.The results of KEGG enrichment showed that those differential proteins were involved in signaling pathways such as completion and coalescence cascades,focal adhesion,arginine biosynthesis,and terpenoid backbone biosynthesis.The complement and coalescence cascades signaling pathway was presumably the target pathway of QWGB in the treatment of POF.In this study,the proteomics technique was used to screen the differential proteins of QWGB in the treatment of POF in mice induced by T.wilfordii glycosides,and they were mainly involved in immune regulation,apoptosis regulation,complement and coagulation cascade reactions,cholesterol metabolism,and steroid hormone production,which may be the main mechanisms of QWGB in the treatment of POF.