苯并[a]芘对雄性褐菖鲉精巢成熟的影响

本实验采用雄性褐菖鲉作为实验对象,腹腔注射0.5×10-6、1×10-6、5×10-6、10×10-6(m/m)苯并[a]芘,7d后苯并[a]芘处理组精巢睾酮水平和睾酮与17β-雌二醇的比值都有上升,但都未表现出显著性差异(P>0.05).腹腔注射50×10-6(m/m)苯并[a]芘,11d后取出精巢,常规石蜡切片,结果显示雄性褐菖鲉的精巢,精小叶腔和输出管中精子密集程度明显增加,次级精母细胞减少,精巢壁厚度变薄.结果提示,苯并[a]芘对褐菖鲉具有类雄激素效应.     

鲈鱼胆汁中芘及代谢产物含量与水体中芘含量的关系

运用GC—MS的方法和荧光分光光度法，研究鲈鱼分别暴露于0．1、1．0和10．0μg／dm^3质量浓度芘溶液中7d，水体中芘及其鲈鱼胆汁中芘和1-羟基芘含量的变化,实验结果显示：(1)鲈鱼对水体中的芘具有非常显著的去除作用。(2)随着芘暴露浓度的增大，鲈鱼对芘的代谢去除作用增强。(3)随着暴露时间的延长和芘暴露浓度的增大，胆汁的1-羟基芘浓度递增。(4)鲈鱼胆汁具有较高的芘浓度，对海水中的芘具有较强的富集作用。(5)鲈鱼胆汁的1-羟基芘及芘的浓度与水体芘浓度均具有很好的相关性，对于指示水体的芘污染程度具有一致性；可作为指示水体芘污染程度的生物标志物。     

苯并[a]芘(BaP)对褐菖鲉(Sebasticus marmoratus)肝CYP1A1酶活性、基因表达及蛋白表达的影响

为了了解苯并[a]芘(BaP)对鱼类细胞色素P4501A1(CYP1A1)表达的影响,以褐菖鲉(Sebasticus marmoratus)为实验材料,采用体内实验,研究其在经过不同浓度(0.1、1、10、20、50mg/kg鱼体重量)的BaP诱导后,鱼体肝脏研究CYP1A1基因表达的情况,筛选出后续时间-效应实验中BaP注射的最佳浓度,研究BaP诱导6h、12h、1d、3d、7d后(质量浓度为20mg/kg鱼体重量)鱼体肝脏CYP1A1酶活性、基因表达和蛋白表达的情况。结果表明:剂量-效应实验中,20mg/kg鱼体重量为最佳浓度,此浓度下,基因表达在各组中变化最显著。时间-效应实验中,较空白对照组而言,染毒6h、12h和1d后,EROD酶活性显著增加。3d后开始下降,与对照组相比变化不大,7d后酶活性又发生上调。半定量RT-PCR结果表明,各染毒组与对照组相比,CYP1A1基因表达量都发生了上调,呈现先上升后下降的趋势。其中,6h和12h组相对表达量极显著增加,1d后开始下降且与3d和7d组相比变化不明显。Western blot结果表明,蛋白表达量在染毒12h后表现出显著的诱导效应,随着时间的延长略有回落,但与对照组相比仍有显著性差异。研究表明:BaP对褐菖鲉CYP1A1具有较强的诱导作用。一定质量浓度的BaP注射于褐菖鲉不同的时间后,能诱导褐菖鲉活体EROD酶活性、CYP1A1基因m RNA表达及蛋白表达,并随着时间的延长呈现先诱导后抑制的趋势。这说明BaP作为诱导剂对CYP1A1酶活性和蛋白表达的作用机制可能与调控CYP1A1的转录水平有关。     

苯并（a）芘诱导下岩虫CYP基因的表达特征

Rapid amplification of cDNA ends(RACE) and real-time polymerase chain reaction(RT-PCR) were carried out to analyze the CYP4 gene expression in polychaete Marphysa sanguinea exposed to benzo[a]pyrene(BaP) in this study. The full length of MsCYP4 cDNA was 2 470 bp, and it encoded 512 amino acids. The deduced amino acid sequence showed 47% identity with CYP4 F from frog Xenopus tropicalis and shared high homology with other known CYP4 sequences. To analyse the role of CYP4 in protecting M. sanguinea from BaP exposure, three BaP groups were established: 0.5, 5 and 50 μg/L. Polychaetes were sampled after 3, 7 and 12 d. At 0.5 μg/L, the effect of BaP on MsCYP4 gene expression increased with time prolonged. MsCYP4 gene expression curve showed Ushaped trend with time in 5 and 50 μg/L BaP groups. Therefore, MsCYP4 gene may play an important role in maintaining the balance of cellular metabolism and protecting M. sanguinea from BaP toxicity.     

Immunotoxicity effect of benzo[α]pyrene on scallop Chlamys farreri

The toxic effects of benzo[α]pyrene (B[α]P) at different concentrations (0.1, 0.5, 1, 2.5 and 7.5 μgL⁻¹) on scallop (Chlamy farreri) immune system were studied. The results showed that B[α]P had significant toxic effects on the haemocyte counts, neutral red uptake, phagocytosis, bacteriolytic and antibacterial activity (P<0.05), while the seawater control and acetone control had no significant differences. The haemocyte counts, neutral red uptake, phagocytosis and bacteriolytic activity in all B[α]P treatment groups as well as antibacterial activity in groups of 0.5, 1, 2.5 and 7.5 μgL⁻¹ B[α]P decreased significantly (P<0.05). Some of these indices tended to be stable on the sixth day and others on the ninth day, and the indices showed clear time- and concentration-response to B[α]P. Bacteriolytic activity in 0.1μgL⁻¹ B[α]P treatment group and antibacterial activity in 0.1 μgL⁻¹ and 0.5 μgL- B[α]P treatment groups increased at the beginning of exposure and reached their peaks on day 1 and day 6, respectively. Following that, both activities decreased gradually and became stable after day 9. When all the indices reached stability, they were significantly lower than those in control group (P<0.05), except for antibacterial activity in 0.1 μgL⁻¹ B[α]P treatment group (P>0.05). Thus, B[α] has evident toxic effects on scallop immune system, which supports the view that a relationship exists between pollution and immunomodulation in aquatic organisms.     

高效液相色谱法分析地下水和饮用水中苯并(a)芘

采用液-液萃取、高效液相色谱法测定地下水和饮用水中的苯并(a)芘,建立了适用于全国地下水污染调查项目多环芳烃中苯并(a)芘必测组分的检测方法。标准曲线的线性范围为2.00～80.0ng/L,相关系数为0.9999,方法检出限为1.8ng/L,回收率为93.1%～103.2%,相对标准偏差(RSD,n=7)为6.48%。方法检出限低,精密度好。     

不同浓度苯并（a）芘对杂色鲍抗氧化酶活性的影响

为阐释不同浓度苯并（a）芘[B（a）P]对杂色鲍（Haliotis diversicolor）的毒性效应,以0.02、0.04和0.08 mg/dm^33个质量浓度的B（a）P对杂色鲍进行水体暴露胁迫,7d后检测肌肉、外套膜、鳃、性腺、肾和肝胰腺的超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、谷胱甘肽S-转移酶（GST）和谷胱甘肽过氧化物酶（GPx）活性及还原型谷胱甘肽（GSH）含量.结果显示：0.02 mg/dm^3的B（a）P暴露对杂色鲍抗氧化系统的影响不明显,而0.04、0.08 mg/dm^3的B（a）P则显著抑制了SOD、CAT和GPx酶活性以及GSH含量,同时对GST酶活性具有显著的诱导作用;实验还发现,不同组织抗氧化酶活性对B（a）P胁迫的敏感性响应存在较大差异,其中鳃、肾脏和肝胰腺的敏感性响应明显高于肌肉和外套膜.上述结果表明,苯并（a）芘暴露对杂色鲍抗氧化酶活性的影响具有明显的剂量-效应关系及组织差异性,杂色鲍的鳃、肾脏或者肝胰腺的抗氧化酶响应多环芳香烃胁迫的更敏感,可作为B（a）P污染的生物标志物.     

苯并芘对褐菖鲉肝细胞DNA交联的影响

以褐菖鲉（Sebastiscus marmoratus）为实验材料,设定苯并芘[B（a）P]的注射剂量梯度分别为0.00、0.05、0.50、5.00 mg/kg,分别于实验的第0、3、6天对每一条鱼进行腹腔注射,于第1、4、7天取样,测定胆汁中的B（a）P及其代谢物含量.采用荧光检测法检测B（a）P染毒后褐菖鲉肝细胞DNA-DNA交联（DDC）交联系数的变化,并用KC l-SDS沉淀法检测DNA蛋-白质交联（DPC）交联系数的变化.结果表明：（1）经注射染毒后鱼体胆汁中的B（a）P及其代谢物含量蓄积程度存在着明显的剂量效应和时间效应;（2）B（a）P的注射剂量分别为0.05、0.50 mg/kg时各时间点褐菖鲉肝细胞DDC交联系数均无显著增大（p〉0.05）,在5.00 mg/kg剂量组,随着时间延长DDC交联系数逐渐增大,第4天即有显著差异（p〈0.05）,第7天差异极显著（p〈0.01）;（3）B（a）P诱导DPC的趋势与DDC类似,但0.5 mg/kg剂量组在第7天DPC交联系数即出现显著增大（p〈0.05）.这表明B（a）P对DPC的诱导高于DDC.这为进一步研究B（a）P的遗传毒性提供了依据,并为探讨B（a）P致癌的作用机理提供了新的信息.     

The genotoxic effects of benzo[a]pyrene and methamidophos on black porgy evaluated by comet assay

In this study, two common pollutants （benzo[a]pyrene and methamidophos） in marine environment were tested by comet assay for their inducement of in vivo genotoxic effect to the blood cells of black porgy （ Acanthopagrus schlegeli）. The fish was exposed to 2 μg/L of benzo[a]pyrene （BaP） and methamidophos, and their mixture. The assay was performed on whole blood at 2 h, 5 h, 24 h and 96 h exposure intervals. A significant increase in DNA damage was observed in each treatment with the pollutants. Additive effect of BaP and methamidophos was also found in the experiment. However, the decrease ratios of DNA damage for 5 h and 96 h exposure interals compared with 2 h and 24 h exposure ones, respectively, were noticed. This phenomenon may be explained by the function of repairing process via enzyme cytochrome P450 in the animal. Evidence of the genotoxicity of organophosphorus pesticides （OPs） and polynuclear aromatic hydrocarbons （PAHs） on marine fish are discussed in this paper.