基于转录组学分析左归丸治疗环磷酰胺诱导卵巢早衰大鼠的分子机制  

Molecular Mechanism of Zuogui Pills in the Treatment of Cyclophosphamide-Induced Premature Ovarian Failure in Rats Based on Transcriptomics Analysis

作　　者：焦存 芦现杰[1] 张梅红[1] 谷万里[1] JIAO Cun;LU Xianjie;ZHANG Meihong;GU Wanli(Liaocheng People's Hospital,Liaocheng 252000 Shandong,China;Liaocheng People's Hospital Affiliated to Shandong University,Liaocheng 252000 Shandong,China)

机构地区：[1]聊城市人民医院,山东聊城252000 [2]山东大学附属聊城人民医院,山东聊城252000

出　　处：《中药新药与临床药理》2023年第3期328-334,共7页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(82004402);山东省中医药科技发展计划项目(2019-0893)。

摘　　要：目的 探讨左归丸对环磷酰胺诱导的卵巢早衰(premature ovarian failure,POF)大鼠的治疗作用,通过转录组测序(RNA-seq)技术分析其分子机制。方法 腹腔注射环磷酰胺复制卵巢早衰大鼠模型。15只大鼠随机分为对照组、模型组和左归丸组。对照组及模型组给予生理盐水灌胃,左归丸组给予2 g·kg^(-1)·d^(-1)左归丸灌胃,连续灌胃4周。ELISA检测各组血清促卵泡激素(FSH)、雌二醇(E_(2))和抗缪勒管激素(AMH)水平,比较各组卵巢湿质量。利用RNA-seq技术检测各组卵巢组织中基因表达谱,筛选目标差异表达基因(differentially expressed genes,DEGs),对DEGs进行基因本体(GO)和京都基因与基因组百科全书(KEGG)富集分析及蛋白互作网络(protein-protein interaction,PPI)分析。结果 与对照组比较,模型组FSH水平升高(P<0.05),E_(2)和AMH水平下降(P<0.05),卵巢湿质量降低(P<0.05);与模型组比较,左归丸组FSH水平下降(P<0.05),AMH水平升高(P<0.05),E_(2)水平和卵巢湿质量的差异无统计学意义(P>0.05)。RNA-seq结果显示,模型组与对照组DEGs 562个,左归丸组与模型组DEGs 393个,左归丸组逆转模型组DEGs 64个。GO富集分析到370个条目,其中生物过程284条,细胞组分40条,分子功能46条。KEGG富集分析主要在卵巢类固醇合成、催乳素信号通路和PPAR信号通路。PPI分析显示,核心基因为EphA5、Acta1、Ros1、Aldh1a1、Cdh1和Cyp19a1。结论 左归丸能够有效改善环磷酰胺诱导卵巢早衰大鼠的血清性激素水平,其分子机制可能与调控类固醇激素合成相关通路及基因有关。Objective To explore the therapeutic effect of Zuogui Pills on cyclophosphamide(CTX)-induced premature ovarian failure in rats and analyze its molecular mechanism by transcriptome sequencing(RNA-seq)technology.Methods The rat model of premature ovarian failure was established by intraperitoneal injection of CTX.Fifteen rats were randomly divided into control group,model group and Zuogui Pills group. The control group and the model group were treated with normal saline, and rats in the Zuogui Pills group were administered intragastrically with 2 g · kg^(-1)· d^(-1)Zuogui Pills for 4 consecutive weeks. The serum levels of follicle-stimulating hormone(FSH),estradiol(E_(2)),and anti-Müllerian hormone(AMH)in each group were detected by ELISA,and the ovarian wet weight in each group was compared. RNA-seq technology was used to detect gene expression profiles in ovarian tissue of each group,and differentially expressed genes(DEGs)were screened. Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis,and protein-protein interaction(PPI)analysis were performed on the DEGs.Results As compared with the control group,the level of FSH in the model group increased(P<0.05),the levels of E_(2)and AMH decreased(P<0.05),and the ovarian wet weight decreased(P<0.05). As compared with the model group,the level of FSH in the Zuogui Pills group decreased(P<0.05),the level of AMH increased(P<0.05),and there was no significant change in the level of E_(2) and ovarian wet weight(P>0.05). RNA-seq results showed that there were 562 DEGs between the model group and control group,393 DEGs between the Zuogui Pills group and model group,and 64 reversed DEGs between the Zuogui Pills group and model group. GO enrichment analysis revealed 370 items,including 284 items of biological process(BP),40 items of cellular component(CC),and 46 items of molecular function(MF). KEGG pathway enrichment analysis mainly focused on ovarian steroidogenesis, prolactin signaling pathway, and PPAR signaling pathway. PPI analy

关 键 词：左归丸 卵巢早衰 转录组测序 环磷酰胺 分子机制 差异表达基因 大鼠 

分 类 号：R285.5[医药卫生—中药学]