联合维生素对邻苯二酸二-（2-乙基己基）酯所致睾丸发育异常的干预作用

Interventional effect of combined vitamin on testicular dysplasia induced by di-（2-ethylhcxyl） phthalate

目的探讨联合应用维生素E和维生素C对邻苯二甲酸二（2-乙基己基）酯所致的睾丸发育异常的干预作用。方法将新生SD大鼠分为5组，即玉米油对照组、邻苯二酸二-（2-乙基己基）酯（DEHP）暴露组、维生素E干预组、维生素C干预组以及联合维生素E和维生素C干预组。药物剂量：DEHP为500 mg/kg、维生素E为200 mg/kg、维生素C为100 mg/kg，每日灌胃，连续处理30 d，6 d后麻醉并处死，测量肛门生殖器距离和肛门生殖指数，HE染色观察睾丸组织病理形态改变情况，酶联免疫吸附测定法（ELISA）检测睾丸组织睾酮含量，Western blot检测SOD及Nrf2的蛋白表达水平。结果与对照组相比，DEHP暴露组和维生素C干预组的肛门生殖指数明显降低（P〈0.05），联合干预组的肛门生殖器距离明显增大（P〈0.05）。HE染色发现DEHP暴露后，大鼠睾丸曲细精管结构紊乱，失去极性，上皮严重萎缩，生精细胞数量极少并且脱落，间质增生明显；维生素E或维生素C干预组的睾丸组织形态也有一定程度破坏，生精细胞数量减少，间质部分增生；联合干预组的睾丸曲细精管结构保存尚好。ELISA结果显示联合干预组的睾丸组织中睾酮含量明显增加（P〈0.05）。DEHP暴露组的SOD和Nrf2表达量明显增高（P〈0.05），其中联合干预组的SOD表达水平明显低于其余各组（P〈0.05），而Nrf2在其余各组中差异没有统计学意义（P〉0.05）。结论联合应用维生素E和维生素C较单独使用其中一种抗氧化剂能够更有效地减轻机体氧化应激压力从而改善环境内分泌干扰物引起的睾丸发育异常，缓解生殖细胞坏死，其中Nrf2信号通路可能起到重要作用。

ObjectiveTo explore the effect of a combination of vitamins E and C on testicular dysplasia induced by di-（2-ethylhcxyl）phthalate （DEHP） in rats.MethodsNeonatal Sprague-Dawley rats were randomly divided into five groups of corn oil control, DEHP exposure, vitamin E, vitamin C and combined vitamins E ＆ C. Gastric administration started from PND1 to PND30.Drug doses： DEHP 500 mg/kg, vitamin E 200 mg/kg, vitamin C 100 mg/kg.At PND36, after anesthetizing, anogenital distance （AGD） was measured for calculating anogenital index （AGI） and hematoxylin eosin staining performed for observing testicle histomorphological change.Enzyme-linked immunosorbent assay （ELISA） was employed for detecting the concentrations of testosterone.And SOD and Nrf2 protein expressions in testes were detected by Western blot.ResultsCompared with control group, AGD and AGI decreased significantly in DEHP exposure and vitamin C groups, but were markedly higher in combined group （P〈0.05）. Subsequently, the structure of seminiferous tubule was disorderly in DEHP group.Polarity became lost, epithelium atrophied severely and the number of spermatogenic cells decreased greatly with exfoliation and interstitial hyperplasia; Morphology of testicular tissue in vitamin E or vitamin C group also had a varying degree of damage, the number of spermatogenic cells decreased and interstitial partial hyperplasia; The testis lumen structures of combined group were well preserved.ELISA showed that testosterone concentration was higher obviously in combined group.The expressions of SOD and Nrf2 were up-regulated markedly in DEHP group （P〈0.05） and SOD expression decreased sharply in combined group （P〈0.05）. Nrf2 had no statistical difference among other groups.ConclusionsA combination of vitamins E and C may be more effective in reducing oxidative stress, improving testicular dysplasia caused by environmental endocrine disruptors and alleviating cell necrosis when compared with using one kind of antioxidants.And Nrf2 signaling pathway may play an important role.