滋肾育胎丸防治早发性卵巢功能不全的转录组和生物信息学研究

Research on Transcriptome and Bioinformatics of Zishen Yutai Pills for Prevention and Treatment of Premature Ovarian Insufficiency

目的:通过转录组与生物信息学结合解析滋肾育胎丸防治早发性卵巢功能不全的关键机制。方法:滋肾育胎丸对早发性卵巢功能不全小鼠模型行预防给药,采用二代高通量测序技术对模型组和给药组小鼠卵巢组织行转录组测序,应用基因本体(GO)富集分析和京都基因和基因组百科全书(KEGG)行生物功能富集分析,通过蛋白质-蛋白质相互作用(PPI)分析方法预测靶基因。采用实时荧光定量PCR(qRT-PCR)技术验证转录组预测靶基因结果的一致性。结果:给药组较模型组,雌激素、抗米勒管激素和卵巢系数升高,组织中不同发育阶段卵泡数目增加;转录组学结果显示,2组间差异表达基因共57个。GO富集分析发现,差异表达基因涉及调控补体和凝血级联反应等通路;KEGG富集分析发现,差异表达基因涉及胆固醇代谢和脂肪消化与吸收等信号通路。PPI与KEGG富集通路结果中重合基因有Apob、Fga、Plg、Fgb、Apoh,是潜在的靶基因;qRT-PCR验证发现,滋肾育胎丸明显升高Fga和Fgb mRNA表达,降低Apob、Plg、Apoh mRNA表达。结论:滋肾育胎丸可通过Fga、Plg、Fgb调控补体和凝血级联反应通路,Apob和Apoh调控胆固醇代谢和脂肪消化与吸收,进而发挥防治早发性卵巢功能不全作用。

Objective:To explore the pivotal mechanisms of Zishen Yutai Pills for the prevention and treatment of premature ovarian insufficiency(POI)based on transcriptome and bioinformatics.Methods:The POI mice models were administered with Zishen Yutai Pills(ZYP)as preventative medicine.The transcriptomes of ovarian tissue from the model and the ZYP treatment groups were sequenced by the second-generation high-throughput sequencing platform.Gene Ontology(GO)enrichment analysis and the Kyoto Encyclopedia of Genes and Genomes(KEGG)were used for biological functional enrichment analysis,and target genes were predicted by protein-protein interaction(PPI)analysis.The consistency of the predicted target gene results of the transcriptome was verified by real-time fluorescent quantitative PCR(qRT-PCR).Results:Compared with the model group,the estradiol,anti-Müllerian hormone,and ovarian coefficient increase in the ZYP treatment group,and the number of follicles in tissue in different developmental stages increases.Transcriptomics results show that there are 57 differentially expressed genes(DEGs)between the two groups.GO analysis shows that DEGs are involved in regulating complement and coagulation cascade reaction pathways,while KEGG analysis finds that DEGs are involved in cholesterol metabolism,fat digestion and absorption,and other signaling pathways.The overlapped genes,Apob,Fga,Plg,Fgb,and Apoh,between PPI and KEGG analysis results are predicted as the potential target genes.qRT-PCR results show that ZYP significantly increases the mRNA expressions of Fga and Fgb and decreases the mRNA expressions of Apob,Plg,and Apoh.Conclusion:ZYP can regulate complement and coagulation cascade reaction pathway through Fga,Plg,and Fgb and cholesterol metabolism and fat digestion and absorption through Apoh and Apob,so as to prevent and treat POI.