氧化应激诱导的自噬与隐睾症及畸形精子症的相关性研究

Studies of Relationship Between Oxidative Stress-induced Autophagy and Cryptorchidism or Teratozoospermia

为了分析氧化应激诱导的自噬与隐睾症及畸形精子症的相关性,首先采用高通量的qPCR array 技术检测44 个自噬相关基因在隐睾症患者和生育力正常男性的睾丸组织中的表达差异,筛选出11 个表达差异显著的自噬相关基因;然后用不同浓度过氧化氢(hydrogen peroxide, H2O2)分别处理小鼠精原细胞系GC-1 spg和睾丸支持细胞系TM4,通过3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐[(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, MTT)]分析H2O2处理对细胞增殖活性的影响,并采用实时荧光定量PCR (real-time quantitativePCR, qRT-PCR)检测11 个差异表达的自噬相关基因在细胞氧化应激条件下的表达;最后利用GEO (GeneExpression Omnibus)数据库分析自噬相关基因与畸形精子症的关系。通过综合比较发现, BCL2L1、EIF2AK3 在隐睾症、畸形精子症及氧化应激过程中均表达上调,表明这两个基因与男性不育的发生有关,推测其作用机制可能是通过响应氧化应激信号来实现。

To analyze the relationship between oxidative stress-induced autophagy and cryptorchidism or teratozoospermia, the high-throughput quantitative PCR array was performed to detect the expression changes of 44 autophagy -related genes in the testicular tissues between patients with cryptorchidism and healthy fertile men. Eleven autophagy genes were selected with significant expression differences. Then the mouse spermatogonial cell line GC-1 spg and testicular sertoli cell line TM4 were respectively treated with different concentrations of hydrogen peroxide (H2O2), and cell viability was analyzed by 3-(4,5-dimethylthiazol-2- yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Next, the real-time quantitative PCR was used to detect the expression of 11 differentially expressed autophagy genes under oxidative stress. Finally, the Gene Expression Omnibus (GEO) database was used to analyze the relationship between autophagy-related genes and terato-zoospermia. By comparison, both BCL2L1 and EIF2AK3 were up-regulated in the testis tissue of patients with cryptorchidism or teratozoospermia, or under oxidative stress, indicating that these two genes should be involved in the occurrence of male infertility, and that the mechanism may be related to their responding to oxidative stress.