苯并[a]芘对马氏珠母贝血淋巴细胞DNA损伤的研究

通过单细胞凝胶电泳技术（彗星试验）研究了苯并[a]芘（B[a]P）对马氏珠母贝（Pinctada martensi）血淋巴细胞DNA的损伤。结果显示,在相同暴露时间内,不同质量浓度的B[a]P均能引起马氏珠母贝血淋巴细胞DNA损伤。4μg.L-1 B[a]P处理组DNA含量、彗尾长度和Olive尾矩均最大,分别为34.39%、281.73和60.31μm。随着暴露时间的延长,彗星尾长明显增加,表现出明显的时间-效应关系,而DNA含量和Olive尾矩没有呈现出规律性变化。此外,在不同的染毒时间内,对照组和各处理组之间均存在显著差异（P〈0.05）,3个指标具有很好的一致性。研究表明,彗星试验是检测B[a]P对马氏珠母贝血淋巴细胞DNA损伤的一种有效手段,马氏珠母贝血淋巴细胞DNA损伤可作为指示B[a]P污染的一种有效生物标志物用于早期预警监测。     

Accumulation and phytoavailability of benzo[a]pyrene in an acid sandy soil

Effects of benzo[a]pyrene (B[a]P) on ryegrass (Lolium perenne L.) growth, plant accumulation and dissipiation of B[a]P in a red sandy soil (Hapli-Udic Argosol) were studied in a pot experiment. The plants were grown for 61 days in soil spiked with B[a]P at 0, 12.5, 25 and 50 mg kg⁻¹. Control pots without plants were also set up. Soil extractable B[a]P, plant shoot and root biomass, and concentrations of B[a]P in plant shoots and roots were determined. Ryegrass biomass was increased by addition of B[a]P and root B[a]P concentrations were significantly correlated with B[a]P application rate, but no such correlation was found for shoot B[a]P concentrations. This indicates that B[a]P enhanced the growth of the ryegrass. The extractable B[a]P concentration in the planted soil was significantly lower than that in the unplanted control soil at the rate of 50 mg B[a]P kg⁻¹. This indicates that ryegrass may help to dissipate B[a]P in soil at concentrations over 50 mg kg⁻¹ soil although the mechanism for this is not understood.     

Benzo [a] pyrene concentrations in topsoils of North Bahrain

A comprehensive field study was conducted to determine the background concentrations of benzo [a] pyrene (B[a]P) in soil samples collected at different points on a grid covering most of the northern and middle parts of the Kingdom of Bahrain. The average B[a]P concentration was 108.4?ng?g^?1 in winter and 73.0?ng?g^?1 in summer. The capital city, Manama, and the industrial areas of the country showed the highest levels of B[a]P.     

苯并[a]芘和1-羟基芘诱导人胚胎干细胞分化心肌细胞ROS、CYP基因表达和DNA损伤

多环芳烃(PAHs)化合物中的苯并[a]芘和PAHs暴露检测标志物1-羟基芘与心脏功能障碍有关,但其生物学机制尚不清楚。为研究苯并[a]芘和1-羟基芘对心脏的毒性作用,基于人胚胎干细胞分化心肌细胞(hESC-CM)研究了苯并[a]芘和1-羟基芘对心肌细胞活性氧(ROS)生成、CYP基因表达和DNA损伤等的影响。结果表明,苯并[a]芘和1-羟基芘对h ESC-CM活性无影响,但能显著增强细胞ROS水平,诱导DNA损伤。此外,苯并[a]芘还能诱导细胞线粒体促凋亡基因的表达。研究表明,苯并[a]芘和1-羟基芘能通过诱导氧化应激和DNA损伤事件导致h ESC-CM损伤,在一定程度上解释了多环芳烃暴露导致心脏疾病的分子机制。     

The synergistic interaction of Nickel(II) with dna damaging agents

In this report we present several examples in which nickel(II) in combination with DNA damaging agents caused an enhanced or synergistic biological response using several different endpoints. These examples of Ni(II) toxicity represent several approaches designed to understand the genotoxicity of Ni(II) as well as several other metal ions. They are discussed in this report as a partial basis for our hypothesis that Ni(II) may alter the cellular processing of DNA damage at some common point in the pathway for DNA repair of several different agents. In cultured Chinese hamster cells DNA damage by Ni(II) ions was not readily demonstrated by the method of alkaline elution, but pretreatment of cells with Ni(II) before X‐irradiation produced an enhanced amount of strand breaks compared to the amount produced by X‐rays alone. A synergistic enhancement of cell transformation of Syrian hamster embryo cells was observed for combined treatments of Ni(II) and benzo(a)pyrene. The nickel enhancement of mutagenesis by ultraviolet light was demonstrated for the bacterial gene gpt stably integrated into the Chinese hamster V79 genome.     

Genotoxic assessment of anthracene and benzo [a] pyrene to milkfish Chanos chanos

Mutagenic and genotoxic effects of polycyclic aromatic hydrocarbons, anthracene and benzo [a] pyrene (BaP), in milkfish Chanos chanos were determined using micronucleus (MN) test and comet assay (CA). Distinct mean frequencies of nuclear abnormalities such as MNs; binucleated micronuclei, nuclear bud, and fragmented apoptotic cells were measured. Significant increase in DNA damage with five classes of damage level was observed and expressed in terms of arbitrary unit (AU). Mean frequencies of total nuclear abnormalities were 0.5?±?0.25 cells in control; 0.67?±?0.33 cells in solvent control; 70?±?9.60 cells in 0.176?mg?L^?1 anthracene, and 91.83?±?6.25 cells in 0.031?mg?L^?1 BaP. The greatest DNA damage of 170AU was observed in 0.176?mg?L^?1 anthracene-exposed group and 182AU was observed in 0.031?mg?L^?1 BaP-treated fish. This study confirmed that the CA and MN assays are useful tools in determining potential genotoxicity of water pollutants and might be appropriate as a part of monitoring program.     

Distribution of benzo[a]pyrene in discrete regions of rat brain tissue using light microscopic autoradiography and gamma counting

In order to study the distribution of benzo[a]pyrene (B[a]P) in rat brain, Sprague-Dawley male rats, 21-day-old, weighted 40–50 g were given a single intravenous (iv) injection of 3.7 × 10⁵ Bq kg^?1 of ¹⁴C-B[a]P. Rats were sacrificed at 1, 6, 12, 24, or 48 h after the administration of radiolabeled B[a]P. Light microscopic autoradiography and gamma counting were used. Gamma counting and light microscopic autoradiography showed that ¹⁴C-B[a]P localized in hippocampus at 1 h, in cerebral cortex at 6 h, and corpus striatum at 24 h after treatment. After B[a]P administration, silver grains rose with time reaching a peak at 24 h and sharp decrease at 48 h. The silver grains distribution was higher in neurons than in gliocytes. Thus, B[a]P penetrated the blood-brain barrier and distributed in various regions of rat brain.     

Mutagenicity of polyaromatic hydrocarbons by chemical models for cytochrome P450 in Ames assay

DNA damage is an important step in carcinogenesis. The Ames assay is a short-term screening of carcinogens that induce DNA damage. Most carcinogens require enzymatic activation through oxidation by cytochrome P450 (CYP450) in the presence of S9 mix. A combination of iron (Fe)(III) porphyrin and an oxidant is also able to oxidize compounds as an alternative metabolic pathway to CYP450. Previously it was reported that a chemical model containing a water-soluble 5,10,15,20-tetrakis(1-methylpyridinium4-yl)porphyrinatoiron(III) chloride (4-MPy) and tert-butyl hydroperoxide (t-BuOOH) activated aromatic amines and amides. In this study, a chemical model composed of an Fe porphyrin, water-insoluble 5,10,15,20-tetrakis(pentafluorophenyl)porphyrinatoiron(III) chloride (F₅P) or water-soluble 4-MPy was optimized with an oxidant – t-BuOOH, magnesium monoperoxyphthalate (MPPT), or iodosylbenzene (PhIO). Subsequently the mutagenicity of benzo[a]pyrene (B[a]P) and chrysene in Salmonella typhimurium TA strains was compared. B[a]P was activated by a combination of F₅P or 4-MPy plus MPPT or PhIO in S. typhimurium TA1538. The B[a]P-induced mutagenicity with F₅P plus oxidant was higher than 4-MPy plus oxidant. Mutagenicity of chrysene, a tetracyclic aromatic hydrocarbon, was not detected in the presence of F₅P/PhIO in S. typhimurium TA98, but was activated in the presence of F₅P/MPPT. The F₅P/MPPT activated other polyaromatic hydrocarbons (PAH) in the S. typhimurium TA98 assay including dibenz[a,c]anthracene, dibenz[a,h]anthracene, 3-methylcholanthrene, and benzo[a]anthracene. The results indicated that the F₅P/MPPT was the most efficient model for detecting PAH-induced mutagenicity in the Ames assay.     

DNA damage in mouse organs and in human sperm cells by bisphenol A

Abstract

The in vivo genotoxic potential of bisphenol A using the comet assay in mice and in human sperm cells in vitro without metabolizing enzymes was studied. Male mice were exposed by oral gavage to the following doses of bisphenol A (0 125, 250 and 500?mg/kg body weight). DNA damage was investigated in liver, kidney, testes, urinary bladder, colon and lungs cells. In testicular cells, a significant increase in DNA strand breaks was observed in the lowest, but not in the medium or highest dose groups. Histopathological investigation of the testicular samples did not show any treatment dose-related effects. No DNA strand breaks were observed in any of the other investigated tissues. In human sperm cells in vitro, bisphenol A did not induce DNA strand breaks.     

Degradation of Benzo[a]Pyrene in Soil with Arbuscular Mycorrhizal Alfalfa

Mycorrhizal and non-mycorrhizal alfalfa (Medicago sativa) was grown in pots containing soil artificially contaminated with various levels of benzo[a]pyrene (B[a]P)(0, 1, 10 and 100 mg kg^–1). Soil and plants were sampled after 30, 40, 50, 60 and 90 days and compared with unlanted pots. The percentage of mycorrhizal root length colonized by Glomus caledoniun was not significantly affected by the addition of B[a]P up to 10 mg kg^–1 but was significantly lower at 100 mg kg^–1B[a]P compared with low concentrations (p < 0.05). There was no difference in soil polyphenol oxidase and dehydrogenase activity among the controls and applications of 1 and 10 mg kg^–1 of B[a]P. However, enzyme activities were significantly higher at 100 mg kg^–1B[a]P compared with the other three treatments, and there was no mycorrhizal effect. Over a period of 90 days the concentration of B[a]P in soil in which alfalfa was grown was significantly lower than in unplanted soil (p < 0.05). Degradation rates of B[a]P added at 1, 10 and 100 mg kg^–1 without G. caledonium were 76, 78 and 53%, and with mycorrhizal inoculation were 86, 87 and 57%. The degradation rate in unplanted soil was significantly lower than in planted soil, and was significantly higher in medium- and low-B[a]P treatments than in the high B[a]P concentration tested. There is a possibility of enhancement phytoremediation of PAHs in rhizosphere soil with arbuscular mycorrhizal fungi.     

Occurrence of and risk posed by highly carcinogenic dibenzo[def,p]chrysene isomers in urban soil

Dibenzopyrene and its isomers are considered the most potent carcinogens of all polycyclic aromatic hydrocarbons tested to date. However, despite public concerns over their deleterious effects, they have not been extensively studied. The occurrence of four highly carcinogenic isomers of dibenzopyrene – dibenzo[def,p]chrysene, dibenzo[a,e]pyrene, dibenzo[a,i]pyrene, and dibenzo[a,h]pyrene in urban soil samples from Shanghai, China, have been determined in this study, as well as that of benzo[a]pyrene and coronene. A total of 14 peaks with ions at m/z 302 were detected by gas chromatography-mass spectrometry. Although the concentrations of benzo[a]pyrene and the sum of the 16 priority polycyclic aromatic hydrocarbons were greater than that of dibenzopyrene and its isomers, the carcinogenic potency of the latter was higher than that of benzo[a]pyrene. The results also indicate that the relative carcinogenic potency of the four dibenzopyrene isomers in the samples is higher than that of the 16 priority polycyclic aromatic hydrocarbons.     

环境污染条件下生物体内DNA损伤的生物标记物研究进展

在正常条件下生物体内的基因组是稳定的;但在环境污染条件下,其DNA容易遭受伤害,主要损伤形式为:碱基改变、脱碱基位点、碱基错配、插入或缺失片段、嘧啶联合、DNA加合物、DNA链断裂、甲基化损伤、DNA链内和链间交联等. 这些受损的DNA对生物细胞产生遗传毒性或细胞毒性. 利用生物标记物进行DNA损伤的检测和定量分析是可行的方法. 本文重点介绍了一些典型的DNA损伤(如DNA加合物、断裂、DNA序列改变等)的生物标记物及其检测方法;认为这些生物标记物在环境污染物的早期诊断和评价方面具有广阔的应用前景. 参32     

Biochemical speciation in chromium genotoxicity

The biochemical speciation of chromium compounds in mammalian cells is discussed with respect to uptake, metabolism, DNA binding and damaging. Whereas soluble hexavalent chromium is taken up rapidly and accumulated intracellularly after its reduction, compounds of trivalent chromium penetrate biomembranes about three orders of magnitude slower. Cr(VI) after its uptake is metabolised by electron donating compounds via Cr(V) to Cr(III) compounds. Chromium from various Cr(III) compounds, but not chromate, binds to chromatin in isolated cell nuclei. The DNA‐protein crosslinks and DNA strand breaks observed in rat liver and kidney after chromate administration are also found in vitro, when Cr(III) compounds (but not chromate) interacts with isolated nuclei. In the Chinese Hamster cell HGPRT mutation assay, three out of four tested Cr(III) complexes were found to be mutagenic. In a direct DNA strand break assay with supercoiled bacteriophage PM 2 DNA, neither chromate nor the four Cr(III) compounds tested caused nicks. However, the combined action of chromate plus glutathione as well as the isolated complex of pentavalent chromium, Na₄Cr(glutathione)₄, did cause DNA breaks. Reactive oxygen species are inferred to be the ultimate DNA nicking agents in this assay. In conclusion there appear to be two mechanisms of chromate genotoxicity; one with direct DNA damage caused by Cr(V) species and one via DNA‐protein crosslinks formed with Cr(III), the final reduction state of chromate.     

Preliminary evidence for the metabolism of benzo(a) pyrene by Plantago lanceolata

Polycyclic aromatic hydrocarbons (PAHs) have long been recognised as potential carcinogens in animals in which biotransformation into reactive metabolites can lead to DNA damage. In animals PAHs metabolism mainly occurs in hepatic microsomes and is associated with the cytochrome p-450 mediated mixed functional oxidase (MFO) system. PAH metabolism in plants has been shown to occur via a similar enzyme system, but has received relatively little attention. This study is looking at how the plant species Plantago lanceolata metabolises benzo(a)pyrene (B(a)P), which is one of the PAHs whose metabolism has been studied extensively in animals. The aim of the work is to establish firstly that the B(a)P is taken up and secondly that it is biotransformed by the plant to products possibly similar to those found in animals. This work is achieved by using C-14-B(a)P along with whole body autoradiography, scintillation analysis and chromatography techniques to locate the B(a)P and its metabolites.     

Polycyclic aromatic hydrocarbons in soils around Guanting Reservoir,Beijing, China

The concentrations of 16 polycyclic aromatic hydrocarbons (∑ 16PAHs) were measured by gas chromatography equipped with a mass spectrometry detector (GC-MS) in 56 topsoil samples around Guanting Reservior (GTR), which is an important water source for Beijing. Low to medium levels of PAH contamination (mean=394.2±580.7 ng g^?1 dry weight (d.w.)) was evident throughout the region. In addition, localised areas of high PAH contamination near steel and cement factories were identified, with ∑ 16PAHs concentrations as high as 4110 ng/g, dry weight (d.w.). There was a significant positive correlation (r²=0.570, p<0.01) between total organic carbon content and ∑ 16PAHs concentrations. Phenanthrene was the predominant compound, accounting for 27.2% of the ∑ PAH concentration, followed by chrysene>pyrene>benzo[a]anthracene≈ benzo[b]fluoranthene≈ benzo[a]pyrene. Four-ring PAH homologues (39%) were dominant. The higher proportion of 4–6 ring homologues, molecular indices, and the spatial distribution of PAH indicated that industrial discharges, incineration of wastes and traffic discharges were the major sources of soil PAHs around the water reservoir.     

Degradation of benzo[a]pyrene in an experimentally contaminated paddy soil by vetiver grass (Vetiveria zizanioides)

A pot experiment was conducted to study the effect of growing vetiver grass on the biodegradation of benzo[a]pyrene (B[a]P) under glasshouse conditions. Plant biomass, microbial biomass C and degradation of B[a]P were determined. B[a]P disappeared faster in the plant treatments than in unplanted controls. Disappearance of B[a]P was accompanied by an increase in soil microbial biomass C. Vetiver grass may promote the biodegradation of B[a]P under flooded conditions by plant roots by stimulating the microbial biomass. Microbial biomass was the main factor affecting dissipation of B[a]P under flooded conditions.     

Multiple-time contamination of benzo(a)pyrene in soils inhibits soil enzymatic activities

Benzo[a]pyrene (B[a]P) is accumulating in soils in a low-dose cumulative manner. The objectives of this study were to investigate the effects of B[a]P on the extractable and available fractions of B[a]P and on soil enzymatic activity using multiple-time superimposed and one-time contamination approaches. Results showed that the contents of B[a]P rapidly decreased in the first 14?d and later decreased slowly from 14 to 56?d in both one-time and multiple-time contamination tests. The contents of B[a]P in the multiple-time contamination test were lower than those in the one-time test. Soil urease, sucrase and dehydrogenase activities were rapidly inhibited in the early stage (14?d) and stimulated during the rest of the incubation, and soil dehydrogenase activity was more sensitive to B[a]P contamination than the other enzymes. High concentrations of B[a]P in soil led to greater inhibition of enzymatic activity than that at low concentrations in the early period of culture. Soil enzyme activities were weakly inhibited in multiple-time compared with in one-time contamination tests and were lower in the subsurface layer than in the surface layer. Our results revealed that the multiple-time superimposed approach might be better than one-time contamination for evaluating B[a]P risk in soil.     

载带苯并[a]芘的纳米碳／二氧化硅颗粒引起大鼠氧化应激作用的研究

通过人工制备载带B[a]P的纳米碳(C)和纳米二氧化硅(SiO2)颗粒,采用气管滴注染毒方式,以7.5mg·kg-1(以体重计)的染毒剂量急性染毒大鼠,观察染毒24小时后载带B[a]P的纳米C/SiO2颗粒对机体产生氧化应激损伤的联合毒性效应.结果表明,在急性染毒后大鼠外周血中反映机体脂质过氧化损伤程度指标的丙二醛(MDA)含量表现为染毒组较对照组显著增加(p<0.05),表明纳米颗粒诱发机体发生了氧化应激反应.在急性染毒后各组大鼠肺泡灌洗液中谷胱苷肽过氧化物酶(GSH-PX)和超氧化物岐化酶(T-SOD)活力与对照组相比显著增加(p<0.05)(载带B[a]P的纳米SiO2组除外);载带B[a]P的纳米SiO2组肺泡灌洗液中GSH-PX活力与对照组相比无显著差异,而与单纯纳米SiO2组和B[a]P组比较显著降低,推测与抗氧化酶的一过性增高有关;载带B[a]P的纳米C组肺泡灌洗液T-SOD活力与其单纯纳米C组和单纯B[a]P组比较显著增加(p<0.05),由此表明载带B[a]P的复合纳米C/SiO2颗粒在致机体氧化损伤效应方面二者存在一定的协同作用.     

RBCA、CLEA及CalTOX模型在苯并[a]芘污染场地健康风险评估中的应用比较

为探讨不同模型对污染场地健康风险评估结果的影响,以苯并[a]芘为例,采用RBCA、CLEA和CalTOX模型对某工业污染场地表层土壤进行健康风险评估,分析了评估结果的差异和原因,同时对模型的主要暴露参数进行了敏感性分析,并推导出基于风险概率分布的土壤修复限值。结果表明,RBCA、CLEA和CalTOX模型计算的苯并[a]芘致癌总风险分别为2.40×10-4、6.32×10-4和7.04×10-6,且经口摄入和皮肤接触2个途径对人体健康造成的危害最大。降解作用是影响CalTOX模型风险评估结果不同于RBCA和CLEA模型的重要因素,3个模型间参数取值及方法学的差异也会导致风险评估结果不同。各模型暴露参数的敏感性排序也有差异。采用基于风险概率分布的方法推导土壤修复限值,RBCA、CLEA和CalTOX模型所得结果分别为0.18、0.08、0.13(不考虑降解作用CalTOX模型)和10.74(考虑降解作用CalTOX模型)mg·kg-1,为各模型直接推导值的1.5～2.6倍。基于风险概率分布的方法可有效降低风险评估过程中参数不确定性的影响,为工业污染场地土壤修复值的制定提供参考。