Abstract

Ageing of the uterine endometrium is a critical factor that affects reproductive success, but the mechanisms associated with uterine ageing are unclear. In this study, we conducted a qualitative examination of age-related changes in endometrial tissues and identified candidate genes as markers for uterine ageing. Gene expression patterns were assessed by two RNA-sequencing experiments using uterine tissues from wild type (WT) C57BL/6 mice. Gene expression data obtained by RNA-sequencing were validated by real-time PCR. Genes expressing the pro-inflammatory cytokines Il17rb and chemokines Cxcl12 and Cxcl14 showed differential expression between aged WT mice and a group of mice composed of 5- and 8-week-old WT (young) animals. Protein expression levels of the above-mentioned genes and of IL8, which functions downstream of IL17RB, were analysed by quantitative immunohistochemistry of unaffected human endometrium tissue samples from patients in their 20s and 40s (10 cases each). In the secretory phase samples, 3,3ʹ- diaminobenzidine staining intensities of IL17RB, CXCL12 and CXCL14 for patients in their 40s were significantly higher than that for patients in their 20s, as detected by a Mann–hitney U test. These results suggest that these genes are candidate markers for endometrial ageing and for prediction of age-related infertility, although confirmation of these findings is needed in larger studies involving fertile and infertile women.

中文翻译：

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鉴定与子宫内膜细胞衰老相关的基因

 抽象的

子宫内膜的老化是影响生育成功的关键因素，但与子宫老化相关的机制尚不清楚。在这项研究中，我们对子宫内膜组织中与年龄相关的变化进行了定性检查，并确定了作为子宫衰老标记的候选基因。使用野生型 (WT) C57BL/6 小鼠的子宫组织通过两次 RNA 测序实验评估基因表达模式。通过 RNA 测序获得的基因表达数据通过实时 PCR 进行验证。表达促炎细胞因子Il17rb以及趋化因子Cxcl12和Cxcl14 的基因在老年 WT 小鼠和由 5 周龄和 8 周龄 WT（年轻）动物组成的小鼠组中表现出差异表达。通过定量免疫组织化学分析 20 多岁和 40 多岁患者（各 10 例）未受影响的人子宫内膜组织样本中上述基因和 IL17RB 下游功能的 IL8 的蛋白表达水平。在分泌期样本中，曼-希特尼U检验检测发现，40多岁患者的IL17RB、CXCL12和CXCL14的3,3′-二氨基联苯胺染色强度显着高于20多岁患者。这些结果表明，这些基因是子宫内膜衰老和预测与年龄相关的不孕症的候选标记，尽管这些发现需要在涉及生育和不孕女性的更大规模研究中得到证实。