广西医学

目的探讨膳食中常见反式脂肪酸——反式油酸致少弱精子症的作用机制。方法通过ProTox和ADMETlab平台预测反式油酸的毒性等级和毒性作用通路。通过ChEMBL、SEA、STITCH、SwissTargetPrediction、TargetNet数据库/平台筛选反式油酸的潜在作用靶点，通过CTD、GeneCards®、OMIM®数据库获取少弱精子症相关靶点，取两者的交集靶点。针对交集靶点，通过Metascape平台进行功能和通路富集分析，基于STRING数据库和Cytoscape软件构建蛋白⁃蛋白相互作用网络并筛选核心靶点。利用AutoDock Vina 软件对核心靶点蛋白与反式油酸进行分子对接分析。结果反式油酸的毒性等级为2级，主要通过Tox21⁃应激反应通路中的核转录因子红系2相关因子2/抗氧化响应元件和热休克反应元件，以及Tox21⁃核受体信号通路中的过氧化物酶体增殖物激活受体γ发挥其毒性作用。共获得36个交集靶点，功能和通路富集分析结果显示，交集靶点主要作用的生物学过程为对类固醇激素的反应、细胞对激素刺激的反应等，主要富集的细胞组分包括神经元细胞体、细胞体、染色体区域等，主要涉及的分子功能主要包括类固醇结合、核受体活性等，主要涉及p53、低氧诱导因子1及肿瘤坏死因子（TNF）信号通路。共筛选得到6个核心靶点，分别为Caspase⁃3（CASP3）、雄激素受体（AR）、核受体亚家族3组C成员1、Erb⁃B2受体酪氨酸激酶2、前列腺素内过氧化物合酶2（PTGS2）、雌激素受体2。分子对接分析结果提示，反式油酸与CASP3（结合能=-7.17 kcal/mol）具有强烈结合活性，与AR（结合能=-5.78 kcal/mol）和PTGS2（结合能=-5.16 kcal/mol）具有较好结合活性。结论反式油酸可能通过激活CASP3⁃p53凋亡相关信号通路诱导生精细胞过度凋亡，并经CASP3⁃PTGS2⁃TNF信号通路触发炎症反应，最终导致精子发生障碍，引发少弱精子症。

Objective To investigate the mechanism of trans⁃oleic acid, a common trans⁃fatty acid in the diet, on oligoasthenospermia. Methods The toxicity grade and toxic effects of trans⁃oleic acid were predicted using the ProTox and ADMETlab platforms. Potential effect targets of trans⁃oleic acid were screened through the ChEMBL, SEA, STITCH, SwissTargetPrediction, and TargetNet databases/platforms. Targets related to oligoasthenospermia were retrieved from the CTD, GeneCards®, and OMIM® databases for acquiring the intersection targets. For these intersection targets, functional and pathway enrichment analyses were conducted via the Metascape platform. A protein⁃protein interaction network was constructed based on the STRING database and Cytoscape software for screening core targets. Molecular docking analysis between core target proteins and trans⁃oleic acid was performed using AutoDock Vina software. Results The toxicity grade of trans⁃oleic acid was grade 2, which was mainly exerted through the nuclear factor erythroid 2⁃related factor 2/antioxidant response element and heat shock response element within the Tox21⁃stress response pathway, as well as peroxisome proliferator⁃activated receptor γ within the Tox21⁃nuclear receptor signaling pathway. A total of 36 intersection targets were identified. Functional and pathway enrichment analysis results indicated that the primary biological processes involving these intersection targets were response to steroid hormone and cellular response to hormone stimulus. Key enriched cellular components included neuronal cell body, cell body, and chromosomal region. Major molecular functions primarily involved steroid binding and nuclear receptor activity. The targets were mainly associated with the p53 signaling pathway, hypoxia⁃inducible factor 1 signaling pathway, and tumor necrosis factor (TNF) signaling pathway. Six core targets were screened as follows: Caspase⁃3 (CASP3), androgen receptor (AR), nuclear receptor subfamily 3 group C member 1, Erb⁃B2 receptor tyrosine kinase 2, prostaglandin⁃endoperoxide synthase 2 (PTGS2), and estrogen receptor 2. Molecular docking analysis results suggested that trans⁃oleic acid exhibited strong binding activity with CASP3 (binding energy=-7.17 kcal/mol) and favorable binding with AR (binding energy=-5.78 kcal/mol) and PTGS2 (binding energy=-5.16 kcal/mol). Conclusion Trans⁃oleic acid may induce excessive apoptosis of spermatogenic cells by activating the CASP3⁃p53 apoptosis⁃related signaling pathway and trigger inflammatory responses via the CASP3⁃PTGS2⁃TNF signaling pathway, ultimately leading to impaired spermatogenesis and oligoasthenospermia.