甲醛是目前全球最主要的环境污染物之一，已有研究显示甲醛具有雄性生殖毒性，但具体分子机制不明。我们前期研究发现甲醛暴露引发大鼠睾丸自噬水平显著增高，且与生精损伤密切相关。本课题拟在前期基础上，以自噬为突破点，深入探索自噬在甲醛诱发生精损伤中的作用和分子机制。具体策略为：①在甲醛暴露大鼠和生精细胞模型中利用电镜，MDC，Real-time PCR，Western blot等手段观察自噬改变；继而抑制自噬，揭示自噬在生精损伤中的作用；②以AMPK/mTOR/ULK1通路为切入点，利用体内外实验验证调控自噬发生的上游分子开关；③在前期芯片筛查基础上，利用IP，激光共聚焦、生物信息学，ChIP等技术分析自噬是否通过调控SPO11、TEX15基因表达进而导致生精异常，明晰自噬引发生精损伤的下游靶点。本课题将为阐明甲醛雄性生殖毒性奠定基础，为进一步防治甲醛雄性生殖毒性提供新的线索和依据。

Formaldehyde is a ubiquitous environmental pollutant. As an important chemical for global economy, formaldehyde output has reached more than 46 billion pounds annually in worldwide. Over the last 20 years, China has experienced rapid economic growth and a simultaneous rise in demand for formaldehyde. In 2004, China surpassed the United States as the largest formaldehyde producer and consumer in the world. China’s actual FA output has reached a staggering 12000 kt in 2007, about 4000 times the amount five decades earlier. Coinciding with the growth, formaldehyde pollution has also increased considerably in China. A substantial body of evidence that consistently indicated the adverse effects of formaldehyde on male reproduction in recent years. However, the molecular and cellular mechanisms underlying this phenomenon remain elusive. .Autophagy is an evolutionarily conserved self-degradation process which provides a membrane-dependent mechanism for the sequestration and lysosomal turnover of subcellular components and organelles. Increasing evidence demonstrated that deregulated autophagy may critically influence vital cellular processes and thereby may play a critical role in the pathogenesis of many human diseases. However its role and characterization remains unclear in male reproductive injuries..Our earlier study has found that markedly enhanced autophagy occurred in testicular tissues of formaldehyde exposure rats. Moreover, the alterations of autophage were consistent with the spermatogenesis injuries. Therefore, the present study was designed to further take autophagy as the breakthrough point, to investigate the role and molecular mechanisms of autophagy in spermatogenesis injuries induced by formaldehyde exposure. Specific strategies include: ① At first, Transmission electron microscopy, MDC, Real-time PCR and Western blot were used to detect autophagy phenomenon in formaldehyde exposure rat models and various spermatogenetic cell models. Afterward, we will further confirm the role of autophagy in male reproductive toxicity via inhibition of autophagy by using autophagy inhibitors or RNAi technique respectively. ② With AMPK/mTOR/ULK1 signaling pathway as the penetration point, we will verify the key upstream molecular switches of autophagy in formaldehyde exposure rat models and various spermatogenetic cell models by using various techniques such as Real-time PCR，Western blot，HPLC，Pathway inhibors or RNAi etc .③ On the basis of our previous Chip screening, Immunoprecipitation (IP), Laser confocal microscopy, Bioinformatics and ChIP techniques will be used to further analysis whether autophagy regulate the expression of spermatogenesis specific genes such as SPO11 and TEX15 to trigger spermatogenic impairment. These experiments will help us to identify the downstream pathways which autophagy induced spermatogenesis injuries. In conclusion, this study will not only lay the foundation for clarifying the male reproductive toxicity of formaldehyde, but also provide new clues and evidence for prevention and treatment male reproductive toxicity of formaldehyde.