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维生素C和维生素E对多氯联苯(氯丹1254)暴露的成年大鼠睾丸间质细胞中类固醇生成酶mRNA表达的影响。

Effects of vitamins C and E on steroidogenic enzymes mRNA expression in polychlorinated biphenyl (Aroclor 1254) exposed adult rat Leydig cells.

作者信息

Murugesan Palaniappan, Muthusamy Thirupathi, Balasubramanian Karundevi, Arunakaran Jagadeesan

机构信息

Department of Endocrinology, Dr. ALM Post Graduate Institute of Basic Medical Sciences, University of Madras, Taramani Campus, Chennai 600113, India.

出版信息

Toxicology. 2007 Apr 11;232(3):170-82. doi: 10.1016/j.tox.2007.01.008. Epub 2007 Jan 16.

Abstract

Polychlorinated biphenyls (PCBs) are ubiquitous and persistent environmental contaminants that disturb normal endocrine functions including gonadal functions in humans and mammals. The present study was conducted to elucidate the protective role of vitamins C and E against Aroclor 1254-induced changes in Leydig cell steroidogenic acute regulatory (StAR) protein and steroidogenic enzymes mRNA expression. Adult male rats were dosed for 30 days with daily intraperitoneal (i.p.) injection of 2 mg/kg Aroclor 1254 or vehicle (corn oil). One group of rats was treated with vitamin C (100 mg/kg bw day) while the other group was treated with vitamin E (50 mg/kg bw day) orally, simultaneously with Aroclor 1254 for 30 days. One day after the last treatment, animals were euthanized and blood was collected for the assay of serum hormones such as luteinizing hormone (LH), follicle stimulating hormone (FSH), testosterone and estradiol. The serum androgen binding protein was also estimated. Testes were quickly removed and Leydig cells were isolated in aseptic condition. Purity of Leydig cells was determined by 3beta-hydroxysteroid dehydrogenase (3beta-HSD) staining methods. Purified Leydig cells were used for quantification of androgen and estrogen receptors. In addition, total RNA was isolated from control and treated Leydig cells to monitor the steady-state mRNA levels by RT-PCR for StAR protein, cytochrome P(450)scc, 3beta-HSD and 17beta-HSD. Aroclor 1254 treatment significantly reduced the serum LH, FSH, testosterone, estradiol and androgen binding protein. In addition to this, Leydig cell androgen and estrogen receptors were markedly decreased. RT-PCR analysis of StAR mRNA level did not alter Aroclor 1254 treatment while steroidogenic enzymes such as cytochrome P(450)scc, 3beta-HSD and 17beta-HSD mRNAs were drastically decreased in Aroclor 1254 treatment. However, the simultaneous administration of vitamins C or E in Aroclor 1254-exposed rats resulted a significant restoration of all the above-mentioned parameters to the control level. These observations suggest that vitamins C and E have ameliorative role against PCBs-induced testicular Leydig cells dysfunction.

摘要

多氯联苯(PCBs)是普遍存在且持久的环境污染物,会干扰包括人类和哺乳动物性腺功能在内的正常内分泌功能。本研究旨在阐明维生素C和维生素E对Aroclor 1254诱导的睾丸间质细胞类固醇生成急性调节(StAR)蛋白及类固醇生成酶mRNA表达变化的保护作用。成年雄性大鼠每日腹腔注射2 mg/kg Aroclor 1254或溶剂(玉米油),持续30天。一组大鼠同时口服维生素C(100 mg/kg体重/天),另一组口服维生素E(50 mg/kg体重/天),与Aroclor 1254同时给药30天。末次给药后一天,处死动物并采集血液,检测血清激素如黄体生成素(LH)、卵泡刺激素(FSH)、睾酮和雌二醇。同时测定血清雄激素结合蛋白。迅速取出睾丸,在无菌条件下分离睾丸间质细胞。通过3β-羟基类固醇脱氢酶(3β-HSD)染色法测定睾丸间质细胞的纯度。纯化的睾丸间质细胞用于定量雄激素和雌激素受体。此外,从对照和处理后的睾丸间质细胞中提取总RNA,通过逆转录聚合酶链反应(RT-PCR)监测StAR蛋白、细胞色素P450侧链裂解酶(P450scc)、3β-HSD和17β-HSD的稳态mRNA水平。Aroclor 1254处理显著降低了血清LH、FSH、睾酮、雌二醇和雄激素结合蛋白水平。除此之外,睾丸间质细胞雄激素和雌激素受体也明显减少。Aroclor 1254处理对StAR mRNA水平无影响,但细胞色素P450scc、3β-HSD和17β-HSD等类固醇生成酶的mRNA在Aroclor 1254处理后大幅下降。然而,在Aroclor 1254暴露的大鼠中同时给予维生素C或维生素E,可使上述所有参数显著恢复至对照水平。这些观察结果表明,维生素C和维生素E对多氯联苯诱导的睾丸间质细胞功能障碍具有改善作用。

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