Nrf2通路对尿道下裂模型中睾丸Leydig细胞的保护作用及机制研究

Environmental Endocrine Disruptors (EDCs), has been well defined as anti-androgenic effects by interfering with production and functional activity of testosterone. Results from recent epidemiological studies indicate association of phthalate esters exposure with a number of human reproductive disorders including hypospadias, cryptorchidism, and spermatogenesis dysfunction. Oxidative stress and mitochondrial dysfunction in Leydig cells were suggested to contribute to phthalate-induced reproductive disorders. Leydig cell is one of the most important targets of testicular toxicity caused by phthalate esters.Meanwhile, Redox-sensitive NF-E2 related factor-2 (Nrf2) is a key transcriptional factor for regulation of oxidative stress in vivo. Fortunately, we have found that Nrf2 and the downstream phaseⅡenzymes were significantly up-regulated in fetal Leydig cells after utero exposure, which indicates the involvement of Nrf2 in protection against oxidative damage.The purpose of our research was to investigate the molecular toxicological mechanism of DBP by analyzing the changes of Nrf2 nuclear translocation and the downstream anti-oxidative proteins, comparing the different outcomes when the expression of Nrf2 has been regulated by the adenovirus transfection, small RNA interfering and sulforaphane. Furthermore，the protective role of Nrf2 in oxidative damage in Leydig cells will be comfirmed in Nrf2-/-and wild type fetal rats. Our research aims to providing therapeutic evidence for human birth defects caused by endocrine disrupting chemicals.

近年来，尿道下裂、隐睾等出生缺陷发病率上升，研究表明这些疾病与环境内分泌干扰物（EDCs）的暴露与蓄积有关，EDCs可导致机体氧化应激损伤，Leydig细胞是其重要靶细胞；NF-E2相关因子2（Nrf2）是调控机体内源性抗氧化应激的重要转录因子。我们前期研究发现，Nrf2及其下游分子在染毒后胎鼠睾丸Leydig细胞内表达上调，推测Nrf2通路可能参与保护Leydig细胞氧化应激损伤。本课题拟从细胞水平分析Nrf2核转位及其下游分子的表达差异，观察Nrf2过表达、干扰表达及莱菔硫烷诱导处理后Nrf2及其下游分子的变化；进一步建立Nrf2基因敲除胎鼠及野生型胎鼠尿道下裂模型，在动物水平验证上述改变，揭示Nrf2对Leydig细胞氧化应激损伤的保护作用及其机制，阐明EDCs导致尿道下裂的分子机制，为防治EDCs所致出生缺陷提供线索。

近年来，尿道下裂、隐睾等出生缺陷发病率上升，研究表明这些疾病与环境内分泌干扰物（EDCs）的暴露与蓄积有关，EDCs 可导致机体氧化应激损伤，Leydig 细胞是其 重要靶细胞；NF-E2 相关因子 2（Nrf2）是调控机体内源性抗氧化应激的重要转录因子。我们前期研究发现，Nrf2 及其下游分子在染毒后胎鼠睾丸 Leydig 细胞内表达上调，推测 Nrf2 通路可能参与保护 Leydig 细胞氧化应激损伤。本课题拟从细胞水平分析 Nrf2 核转位及其下 游分子的表达差异，观察 Nrf2 过表达、干扰表达及莱菔硫烷诱导处理后 Nrf2 及其下游分子的变化；进一步建立 Nrf2 基因敲除胎鼠及野生型胎鼠尿道下裂模型，在动物水平验证上述改变，揭示 Nrf2 对 Leydig 细胞氧化应激损伤的保护作用及其机制，阐明 EDCs 导致尿道下裂的分子机制，为防治 EDCs 所致出生缺陷提供线索。