邻苯二甲酸二丁酯诱导氧化应激及抑制CYP17a1干扰睾酮合成  

Testicular oxidative stress and downregulation of CYP17a1 indused by di(n-butyl) phthalate inhibit synthesis of testosterone

作　　者：于淼[1] 张林媛[1] 乔佩环[1] 常兵[1] 

机构地区：[1]中国疾病预防控制中心职业卫生与中毒控制所,北京100050

出　　处：《卫生研究》2015年第3期364-370,共7页Journal of Hygiene Research

基　　金：国家自然科学基金(No.30972447;30150003);科技部社会公益研究项目(No.2001DIB0059)

摘　　要：目的探讨邻苯二甲酸二丁酯(DBP)导致睾丸氧化损伤与睾酮合成途径的关系及可能的机制。方法 24只健康4周龄雄性Wistar大鼠随机分为对照组(玉米油)及DBP低、中、高剂量组(80、200和500 mg/kg),每组6只,经灌胃染毒每日1次、连续染毒4周。染毒结束后称量动物体重及睾丸和附睾的重量,用生化方法检测睾丸匀浆中丙二醛(MDA)和活性氧自由基(ROS)的含量,抗氧化酶超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶-1(GPx-1)的活性变化,类固醇合成酶3β-HSD1、17β-HSD3的活性,用放射免疫法测定外周血中睾酮、促黄体生成素(LH)、卵泡刺激素(FSH)和睾丸内的睾酮(T)、雄烯二酮(ASD)的水平,用real timeq PCR方法检测St AR、P450scc、3β-HSD1、17β-HSD3、CYP17a1 mRNA水平的表达变化。结果与对照组相比,高剂量组体重、睾丸重量明显减少(P<0.05);循环血LH、FSH增高(P<0.05),循环血及睾丸内睾酮、睾丸内ASD均降低(P<0.05);MDA和ROS的含量明显升高(P<0.05),SOD、CAT、GPx-1的活性及3β-HSD1的酶活性均明显降低(P<0.05);St AR、P450scc、3β-HSD1、CYP17a1 mRNA降低,17β-HSD3 mRNA升高(P<0.05)。中剂量组循环血LH、FSH增高(P<0.05),睾丸内T和ASD降低(P<0.05);ROS升高(P<0.05),GPx-1的活性降低(P<0.05);3β-HSD1酶活性减少(P<0.05);St AR、P450scc、CYP17a1 mRNA降低(P<0.05)。低剂量组所有指标未见有统计学意义的改变。结论 DBP暴露干扰了大鼠睾丸氧化/抗氧化平衡,降低了GPx-1活性,抑制CYP17a1基因的表达,降低睾丸ASD水平,最终抑制间质细胞内睾酮合成。CYP17a1降低是DBP干扰睾酮合成的毒性作用机制之一。Objective To explore the relationship and possible mechanisms between testicular oxidative injury caused by DBP and the testosterone synthesis pathway. Methods Twenty-four male Wistar male rats （4-wk-old） were randomly divided into 4 groups , three doses of DBP （80, 200 and 500 mg/kg） groups and a vehicle （corn oil） control group, 6 animals each. The rats were respectively administered by garage once a day for four weeks. They were sacrificed after 4 weeks treatment and the body weights, testis, epididymis were weighed, respectively. The oxidation of MDA and ROS, the activity changes of antioxidases SOD, CAT and GPx-1, as well as the activities of steroid synthetases 3β-HSD1, 17β-HSD3 in the testis homogenate were measured by biochemical methods. The levels of testosterone, LH , FSH in peripheral blood and testosterone, ASD in testis were measured by radioimmunoassay. The intensities of expresses of STAR, P450scc, 3β-HSD1,17β-HSD3 and CYP17a1 mRNA were detected by real-time qPCR. Results In 500 mg/kg dose group, the body weights and weigths of testis were decreased obviously （P 〈 0. 05 ）. The concentration of serum LH and FSH was increased, the consentration of serum T, testicular T and testicular ASD was decreased （P 〈 0.05）. The oxidation of MDA and ROS was increased distinctly and the activities of SOD, CAT, GPx- 1 and 3β-HSD1 were reduced （P 〈 0. 05）. STAR, P450scc, 3β-HSD1 and CYP17a1 mRNA were decreased, 17β-HSD3 mRNA was increased （P 〈 0.05）. In 200mg/kg dose groups, LH , FSH level in peripheral blood were increased and ASD level in testis was decreased （ P 〈 0. 05 ）. The oxidation of ROS was increased and activity of GPx-1 and 3β- HSD1 were decreased （P 〈0.05）. STAR, P450scc and CYP17a1 mRNA were decreased （P 〈 0.05）. There were no changes in 80mg/kg group. Conclusion DBP exposure disturbed the balances of oxidation/antioxidation, then result in the decline of GPx-1 activity, CYP17a1 mRNA and ASD level which caused the decrease of testosterone

关 键 词：邻苯二甲酸二丁酯 间质细胞 氧化损伤 CYP17a1 睾酮合成 

分 类 号：R994.6[医药卫生—毒理学] Q343.34[医药卫生—药学]