胶原受体DDR2与非梗阻性无精症的相关性研究  

Study on the Correlation of Collagen Receptor DDR2 with Non-obstructive Azoospermia

作　　者：颜学友 郝耀 李晓阳 张佳惠 陈国栋 张淑雅[1,2] YAN Xueyou;HAO Yao;LI Xiaoyang;ZHANG Jiahui;CHEN Guodong;ZHANG Shuya(School of Basic Medicine,Ningxia Medical University,Yinchuan 750004,China;Key Laboratory of Fertility Preservation and Maintenance of Ministry of Education,Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学基础医学院,银川750004 [2]宁夏医科大学生育力保持教育部重点实验室,银川750004

出　　处：《宁夏医科大学学报》2024年第11期1081-1090,共10页Journal of Ningxia Medical University

基　　金：国家自然科学基金项目(82460294);宁夏卫生健康系统科研课题(2022-NWKY-049);宁夏重点研发计划项目(引才专项)(2022BSB03090);宁夏医科大学特殊人才启动项目(XT2021001)。

摘　　要：目的探究胶原受体DDR2在非梗阻性无精症(NOA)中的作用及其相关机制。方法获取GEO数据库NOA患者睾丸组织基因芯片数据(GSE45885),通过limma差异基因分析与WGCNA分析,检测DDR2在4种不同严重程度NOA之间的表达变化及相关性。通过分析睾丸组织单细胞转录组测序数据,明确DDR2在睾丸中的表达定位、拟时序动态变化及靶标细胞间的通信关系,并使用DDR2^(-/-)小鼠模型验证其对精子发生的调控作用。结果DDR2与严重NOA的发病密切相关,其表达水平随无精症病症加重而逐渐升高,在唯支持细胞综合征(SCOS)患者睾丸组织中表达水平最高(P<0.001);其次为减数分裂前生精阻滞、减数分裂期生精阻滞和减数分裂后生精阻滞。DDR2高表达于睾丸间质细胞(LCs)和管周肌样细胞(PMCs),其表达水平在成纤维来源细胞拟时序中与标记基因Cyp17a1、Myh11和Sbspon相一致,在分化中期和后期细胞高表达,在分化前期细胞中低表达。细胞通信分析显示,DDR2可能通过Col1a1_Col1a2-Itga9_Itgb1/Sdc4等配受体关系参与调控LCs和PMCs细胞间通信。DDR2^(-/-)雄鼠成熟精子精子活率、精子活动总数及有效精子数均显著降低,表现出严重的精子发生障碍。结论DDR2与严重NOA的发病密切相关,且可能通过参与LCs和PMCs细胞间通信而影响精子发生。Objective To investigate the correlation between collagen receptor DDR2 and non-obstructive azoospermia(NOA)and to clarify the localization of its expression in the testis.Methods Testicular tissue chip data(GSE45885)of NOA patients from GEO database were obtained,and the expression changes and correlation of DDR2 among four different severities of NOA were detected by limma differential gene analysis and WGCNA analysis.By analyzing the single-cell transcriptome sequencing data of testicular tissues,the expression localization of DDR2,the pseudo-temporal dynamic changes and the communication relationship between the target cells were clarified.Finally,a DDR2^(-/-)mice model was used to validate the regulatory role of DDR2 on spermatogenesis.Results DDR2 was closely associated with the development of severe NOA,and its expression level increased with the worsening of azoospermia,with the highest level in patients with sertoli cell-only syndrome(SCOS)(P<0.001),followed by meiotic arrest,meiotic arrest,and post-meiotic arrest.DDR2 was highly expressed in testicular leydig cells(LCs)and peritubular myoid cells(PMCs),with expression levels consistent with the marker genes Cyp17a1,Myh11,and Sbspon in the pseudo-temporal order of fibroblast-derived cells,and was overexpressed in mid-and late-stage differentiated cells,and underexpressed in pre-differentiated cells.Analysis of cellular communication revealed that DDR2 might be involved in regulating intercellular communication between LCs and PMCs through the ligand-receptor of Col1a1_Col1a2-Itga9_Itgb1/Sdc4.DDR2^(-/-)male mice exhibited severe spermatogenesis disorders,with significant reductions in sperm viability,total sperm motility,and effective sperm counts.Conclusion DDR2 was closely related to the pathogenesis of severe NOA,and may affect spermatogenesis by participating in the communication between LCs and PMCs.

关 键 词：非梗阻性无精症 胶原受体DDR2 睾丸间质细胞 

分 类 号：R698.2[医药卫生—泌尿科学]