彭思佳,陈宝艳,梁靖谊,潘兆丰,杨云虹,谢秋月,孙晓峰
PENG Sijia,CHEN Baoyan,LIANG Jingyi,PAN Zhaofeng,YANG Yunhong,XIE Qiuyue,SUN Xiaofeng

• 1:广州中医药大学第一临床医学院
• 2:广州中医药大学第一附属医院产科

摘要(Abstract)：

目的 利用生物信息学技术分析妊娠期糖尿病(GDM)中的关键差异基因及胎盘免疫细胞浸润模式并进行实验验证，探讨GDM的发病机制。方法 在基因表达综合数据库(GEO)筛选妊娠期糖尿病基因探针芯片数据集，使用在线分析工具GEO2R从所研究数据集中筛选出差异基因(DEGs)并使用注释文件进行注释，利用STRING及Cytoscape中的CentiScaPe工具进行蛋白质-蛋白质相互作用(PPI)分析，运用Metascape数据库对关键差异基因进行基因本体论(GO)和京都基因与基因组百科全书(KEGG)进行分析，再进一步使用CIBERSORTx工具对人类22种免疫细胞亚型表达矩阵进行去卷积分析，最终得出关键差异基因及免疫细胞浸润模式；收集分娩后的GDM和非GDM胎盘组织各3例，采用qRT-PCR和Western blot对关键差异基因进行验证。结果 共筛选出207个DEGs(P<0.01),其中包括60个上调基因和147个下调基因；PPI结果共获得105个节点和235个基因对，最终选择出补体C3(C3)、人类白细胞抗原C(HLA-C)、HLA-B、C-X-C基序趋化因子10(CXCL10)四个关键基因；KEGG及GO分析结果表明免疫效应、机体防御反应、免疫反应分子介质生成与释放、白细胞分化、同种异体移植排斥通路、抗原加工和呈递等信号途径参与了GDM的发病过程，免疫细胞浸润分析得出单核细胞、M2巨噬细胞在GDM胎盘组织中的浸润程度分别呈下降(P<0.05)和上升趋势(P<0.05);qRT-PCR和Western blot结果显示分娩后的GDM胎盘组织中C3、CXCL10、HLA-B、HLA-C mRNA表达均较非GDM胎盘有组织升高，差异有统计学意义(P<0.05)。结论 GDM胎盘变化中潜在的关键基因为C3、HLA-C、HLA-B、CXCL10,单核细胞、M2巨噬细胞可能在疾病的发病过程中起着重要作用。
Objective To analyze the key differential genes in gestational diabetes mellitus(GDM) and the infiltration pattern of placental immune cells by bioinformatics technology, and carry out experimental verification to explore the pathogenesis of GDM. Methods The gene probe chip data set of gestational diabetes mellitus was screened in the Gene Expression Omnibus(GEO) database. Differential genes(DEGs) were screened from the research data set by using the online analysis tool GEO2 R and they were annotated with annotation files. Protein-protein interaction(PPI) analysis was conducted utilizing the CentiScaPe tool in STRING and Cytoscape. The Metascape database was employed to analyze the key differential genes in the Gene Ontology(GO) and the Kyoto Encyclopedia of Genes and Genomes(KEGG). Further, the CIBERSORTx tool was used to deconvolute the expression matrix of 22 human immune cell subtypes to obtain key differential genes and immune cell infiltration patterns. Three GDM and three non-GDM placental tissues were collected after delivery, and the key differential genes were verified by quantitative real-time polymerase chain reaction(qRT-PCR) and Western blot. Results A total of 207 DEGs were screened(P<0.01), including 60 up-regulated genes and 147 down-regulated genes. PPI results showed that 105 nodes and 235 gene pairs were obtained, and four key genes, complement component 3(C3), human leukocyte antigen C(HLA-C), HLA-B, C-X-C motif chemokine ligand 10(CXCL10), were finally selected. The results of KEGG and GO analysis demonstrated that the immune effect, body defense response, the generation and release of molecular mediators of immune response, leukocyte differentiation, allograft rejection pathway, antigen processing, and presentation and other signal pathways were involved in the pathogenesis of GDM. The immune cell infiltration analysis showed that the infiltration degree of monocytes in GDM placental tissue decreased(P<0.05) and that of M2 macrophages increased(P<0.05). The results of qRT-PCR and Western blot showed that the mRNA expressions of C3, CXCL10, HLA-B, and HLA-C in GDM placental tissue after delivery were higher than those in the non-GDM placenta and the differences were statistically significant(P<0.05). Conclusion The potential key genes in placental changes of GDM are C3, HLA-C, HLA-B, and CXCL10, monocytes and M2 macrophages may play an important role in the pathogenesis of the disease.

关键词(KeyWords)： 生物信息学;妊娠期糖尿病;胎盘;关键基因;免疫浸润
bioinformatics;gestational diabetes mellitus(GDM);placenta;key genes;immune infiltration

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(81603653)

作者(Author): 彭思佳,陈宝艳,梁靖谊,潘兆丰,杨云虹,谢秋月,孙晓峰
PENG Sijia,CHEN Baoyan,LIANG Jingyi,PAN Zhaofeng,YANG Yunhong,XIE Qiuyue,SUN Xiaofeng

DOI: 10.19367/j.cnki.2096-8388.2023.01.005

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