Rao Meng, Zeng Zhengyan, Tang Li, Cheng Guiping, Xia Wei, Zhu Changhong
Department of Reproduction and Genetics, The First Affiliated Hospital of Kunming Medical University, Kunming, China.
Department of Neurology, The First Affiliated Hospital of Kunming Medical University, Kunming, China.
Oncotarget. 2017 Dec 4;8(67):111672-111682. doi: 10.18632/oncotarget.22900. eCollection 2017 Dec 19.
This study aimed to investigate the role of microRNA (miRNA) in heat stress-induced spermatogenic impairment. Testes from 15 adult ICR mice subjected to testicular hyperthermia at 43°C for 30 min and from 15 control mice were collected and pooled into 3 samples. Isolated RNA from these samples was subjected to small RNA high-throughput sequencing, and differentially expressed miRNAs were identified and validated using RT-PCR. The identified miRNAs were further subjected to Gene Ontology and KEGG analyses, which revealed significant enrichment for pathways potentially involved in heat stress-induced spermatogenic impairment. Additionally, a correlation analysis of the relative levels of validated miRNAs with germ cell apoptosis was performed. Of the 11 miRNAs identified as differentially expressed, 8 were validated as consistent with sequencing data. Further analyses suggested that the target genes of those miRNAs were involved in various pathways (e.g., ribosomal, HIF-1, MAPK) that may be critical to heat stress-induced testicular damage. Some identified miRNAs, including miR-449a-3p, miR-92a-1-5p, miR-423-3p, and miR-128-3p, correlated closely with germ cell apoptosis. The study results reveal a detailed miRNA profile of heat stress-induced testicular damage and highlight new and potentially important candidate targets in the process of male infertility.
本研究旨在探讨微小RNA（miRNA）在热应激诱导的生精功能障碍中的作用。收集15只成年ICR小鼠在43°C下进行30分钟睾丸高温处理后的睾丸以及15只对照小鼠的睾丸，并将其合并为3个样本。从这些样本中分离的RNA进行小RNA高通量测序，并使用RT-PCR鉴定和验证差异表达的miRNA。对鉴定出的miRNA进一步进行基因本体论和KEGG分析，结果显示在热应激诱导的生精功能障碍潜在涉及的途径中存在显著富集。此外，对验证的miRNA相对水平与生殖细胞凋亡进行了相关性分析。在鉴定为差异表达的11个miRNA中，8个经验证与测序数据一致。进一步分析表明，这些miRNA的靶基因参与了各种可能对热应激诱导的睾丸损伤至关重要的途径（如核糖体、HIF-1、MAPK）。一些鉴定出的miRNA，包括miR-449a-3p、miR-92a-1-5p、miR-423-3p和miR-128-3p，与生殖细胞凋亡密切相关。研究结果揭示了热应激诱导睾丸损伤的详细miRNA图谱，并突出了男性不育过程中新的潜在重要候选靶点。
