杨梅花色苷抑制重金属损伤睾丸间质细胞的机制研究

项目名称： 杨梅花色苷抑制重金属损伤睾丸间质细胞的机制研究

项目编号： No.31201340

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 膳食与营养

项目作者： 白卫滨

作者单位： 暨南大学

项目金额： 27万元

中文摘要： 重金属对人类的生殖健康造成的威胁日益引起人们重视，前期研究发现重金属可通过氧化损伤途径导致雄性生殖毒性，因此本研究拟"以抗氧化性强的食源性功能因子实现对重金属暴露的雄性生殖毒性的抑制"这一新思路，以矢车菊-3-葡萄糖苷（Cy-3-Glu）为模式化合物，以镉、汞、铅等重金属损伤的雄性睾丸间质（Leydig）细胞为模型，采用流式细胞仪和共聚焦显微镜等手段观察花色苷干预后细胞形态和功能的变化，采用实时荧光RT-PCR、蛋白印迹等技术分析花色苷干预后细胞的线粒体抗氧化系统和睾酮生成途径关键酶在mRNA及蛋白水平表达的变化，阐明花色苷对重金属暴露的Leydig细胞损伤的抑制途径。采用2', 7'-二氯荧光黄双乙酸盐探针和 Rose-Drago 方法分别测定细胞中自由基变化和重金属螯合率，结合抑制途径探讨花色苷的抑制机理。本研究对于利用花色苷预防重金属对人体生殖毒性危害具有重要的理论价值和现实意义。

中文关键词： 重金属；矢车菊素-3-葡萄糖苷；孕酮；线粒体膜电位；ROS

英文摘要： In rencent years, much attention has been paid on heavy metal-induced human reproductive toxicity and it is revealed that heavy metal could induce male reproductive toxicity through oxidative damage pathway in our previews study. So based on the idea of "functional factor with high antioxidative activity in food could inhibit effects of heavy metals-induced male reproductive toxicity", we designed this study to intervene Leydig cells damaged by Cd, Hg and Pb using Cy-3-Glu to clarify the inhibitory approach of anhthocyains to damaged Leydig cells induced by heavy metals though observing the change of cell morphology and function with Flow Cytometry and confocal microscope,the change of mRNA and express of the key enzyme in mitochondria antioxidation system and formation process of testosterone with real-time fluorescence quantitative RT-PCR and Western blot method. Besides, the change of free radical concentration and heavy metal chelate ratio in cells were also investigated using the fluorescent probe 2′,7′-dichlorofluorescin-diacetate and Rose-Drago method respectively, and the results will be combined with inhibit pathway to explore the inhibitory mechanism of anthocyanins. This research is very important for the application of anthcyanins in protecting human from heavy metals induced reproductive toxicity an

英文关键词： heavy metal；Cyanidin-3-glucoside；progesterone；mitochondrial membrane potential；ROS