Noaksson Erik, Linderoth Maria, Bosveld Albertus T C, Balk Lennart
Laboratory for Aquatic Ecotoxicology, Institute of Applied Environmental Research, Stockholm University, S-106 91 Stockholm, Sweden.
Gen Comp Endocrinol. 2003 Dec;134(3):273-84. doi: 10.1016/s0016-6480(03)00267-3.
Endocrine disruption associated with reproductive failure has been reported previously in female perch (Perca fluviatilis) and roach (Rutilus rutilus) from Lake Molnbyggen in Sweden and in female brook trout (Salvelinus fontinalis) from Vadbäcken, a stream emptying into Molnbyggen. Both Molnbyggen and Vadbäcken have been contaminated by toxic leachate from a municipal refuse dump. In this study, female perch were caught in Molnbyggen and the reference lake, Lake Djursjön, to further investigate the endocrine mechanism behind the significant numbers of sexually immature (SIM) female perch in Molnbyggen. Blood plasma analysis of progesterone (P), 17alpha-hydroxyprogesterone (17alpha-OHP), testosterone (T), and 17beta-oestradiol (E2), as well as analysis of brain aromatase activity (P450arom), were carried out. The exceptional high numbers of SIM female perch in Molnbyggen was confirmed in February 1999. In July 1999, at an early stage of oogenesis, perch from Molnbyggen showed significantly decreased gonadosomatic index (GSI) and aromatase activity. The presence of aromatase inhibiting substances in lake sediments were therefore tested in vitro. The aromatase activity was dose-dependently inhibited by clotrimazole, reaching 50% inhibition at a concentration of 0.9 microM. Aromatase inhibiting substances were found both in Molnbyggen and reference sediment extracts, indicating that they were naturally occurring substances and not of anthrophogenic origin. The similar decrease in levels of circulating steroids (P, 17alpha-OHP, T, and E2), aromatase, and GSI therefore suggest that the low aromatase activity is due to down-regulation rather than inhibition. To further investigate the steroidogenesis prior to T, P, and 17alpha-OHP were analysed in perch caught in 1997 and 1998 in Lakes Molnbyggen, Kvarntjärn (downstream), Yxen (upstream), and Djursjön, in female roach caught in Molnbyggen and Djursjön in 1997, and in brook trout caught in Vadbäcken and the reference stream Björntjärnsbäcken in 1998. The absence of differences in P and 17alpha-OHP levels, combined with a significantly lower T level in female perch and roach from Molnbyggen in 1997, could be the result of either increased metabolism and excretion of T, or a disruption downstream of 17alpha-OHP formation. The unaffected P levels and significantly lower 17alpha-OHP levels, together with significantly decreased T and E2 levels, found in adult (>45g) female brook trout from Vadbäcken, further indicated that an altered steroidogenesis downstream of P is one possible mechanism underlying the low T levels and thus the high number of SIM female fish, since too low T levels might be insufficient to activate the brain-pituitary-gonadal axis.
此前有报道称,瑞典莫恩比根湖中的雌性河鲈(Perca fluviatilis)和拟鲤(Rutilus rutilus)以及流入莫恩比根湖的瓦德贝克溪中的雌性溪红点鲑(Salvelinus fontinalis)出现了与生殖失败相关的内分泌紊乱。莫恩比根湖和瓦德贝克溪均受到了来自城市垃圾场有毒渗滤液的污染。在本研究中,在莫恩比根湖和对照湖尤尔松湖捕获了雌性河鲈,以进一步探究莫恩比根湖中大量性未成熟雌性河鲈背后的内分泌机制。对血浆中的孕酮(P)、17α - 羟基孕酮(17α - OHP)、睾酮(T)和17β - 雌二醇(E2)进行了分析,同时也分析了脑芳香化酶活性(P450arom)。1999年2月证实了莫恩比根湖中存在异常大量的性未成熟雌性河鲈。1999年7月,在卵子发生的早期阶段,来自莫恩比根湖的河鲈性腺体指数(GSI)和芳香化酶活性显著降低。因此,对湖泊沉积物中芳香化酶抑制物质的存在进行了体外测试。克霉唑对芳香化酶活性具有剂量依赖性抑制作用,在浓度为0.9微摩尔时达到50%的抑制率。在莫恩比根湖和对照沉积物提取物中均发现了芳香化酶抑制物质,这表明它们是天然存在的物质,而非人为来源。循环类固醇(P、17α - OHP、T和E2)、芳香化酶和GSI水平的类似下降因此表明,低芳香化酶活性是由于下调而非抑制所致。为了进一步研究睾酮合成之前的类固醇生成,对1997年和1998年在莫恩比根湖、夸恩特耶恩湖(下游)、于克森湖(上游)和尤尔松湖捕获的河鲈中的P和17α - OHP进行了分析,对1997年在莫恩比根湖和尤尔松湖捕获的雌性拟鲤以及1998年在瓦德贝克溪和对照溪流比约恩特耶恩斯贝克溪捕获的溪红点鲑中的P和17α - OHP进行了分析。1997年莫恩比根湖中的雌性河鲈和拟鲤中P和17α - OHP水平没有差异,而T水平显著较低,这可能是T代谢和排泄增加的结果,也可能是17α - OHP形成下游的干扰所致。在来自瓦德贝克溪的成年(>45克)雌性溪红点鲑中发现未受影响的P水平和显著较低的17α - OHP水平,同时T和E2水平显著降低,这进一步表明P下游类固醇生成的改变是T水平低以及因此性未成熟雌性鱼类数量多的一种可能机制,因为过低的T水平可能不足以激活脑 - 垂体 - 性腺轴。