硝基芳烃化合物诱发细胞微核的比较研究

利用蚕豆根法细胞和人在淋巴细胞的微核试验对６种硝基芳烃化合物的致突变性进行比较研究，结果表明，邻二硝基苯、间二硝基苯、２，４－二硝基甲苯，２，６－二硝基甲苯，对硝基氯苯和对硝基溴苯均能诱发两咱细胞的微核率增加，除了对硝基溴苯外均具有明显剂量一效应关系。引进标准剂量概念比较５种硝基芳烃人合物的致突变性，结果表明，致突变性强弱依次为２，４－二硝基甲苯〉对硝基氯苯〉间二硝基苯〉２，６－二硝基甲苯〉邻二硝     

Immunotoxicity of bisphenol a to Carassius auratus Lymphocytes and macrophages following in vitro exposure

Bisphenol A （BPA） is the monomer component of polycarbonate plastics and classified as an endocrine disrupting chemical （EDC）. The reproductive toxicity of BPA has been extensively studied in mammals; however, relatively little information is available on the immunotoxic responses of fish to BPA. In this study, we investigated the effects of BPA on the immune functions of lymphocytes and macrophages in Carassius auratus. The effects of BPA were compared with those of two natural steroid hormones, estradiol and hydrocortisone. Proliferation of the two types of cells in response to PHA was measured using colorimetric MTT assay. Macrophage respiratory burst stimulated by Con A was measured using chemiluminescence assay. Results showed that BPA （0.054-5.4 mg/L）, estradiol （0.0002-2.0 mg/L） and hydrocortisone （5-50 mg/L） significantly induced Carassius auratus lymphocyte proliferation while higher doses of hydrocortisone （500-5000 mg/L） appeared to be inhibitory. BPA （0.005-50 mg/L）, estradiol （0.005-800 mg/L） and hydrocortisone （0.005-500 mg/L） markedly enhanced macrophage proliferation, whereas higher doses of BPA （500-1000 mg/L） appeared to inhibit cell proliferation. Furthermore, higher dosage of BPA （50 mg/L） and hydrocortisone （50 and 500 mg/L） suppressed the macrophages respiratory burst while estradiol is stimulative all the doses tested （0.05-500 mg/L）. In conclusion, BPA could have immunotoxicity to Carassius auratus and functional changes of lymphocyte and macrophage in Carassius auratus may be different between low and high dosages.     

Micronucleus frequency in sheep lymphocytes after in vitro exposure to fungicide tolylfluanid

The fungicide tolylfluanid (N - dichlorofluoromethylthio-N′, N - dimethyl - N - p - tolylsulfamide), was investigated by cytokinesis-block micronucleus assay. Tolylfluanid at the lowest concentration (1 × 10^{? 6}mol L^{? 1})did not influence significantly the frequency of micronuclei in sheep lymphocyte cultures in comparison with control (32.33 ± 3.51/1000 binucleated cells versus 30.33 ± 2.82/1000 binucleated cells in dimethylsulfoxide (DMSO) control, P = 0.44). Higher tolylfluanid concentrations (1 × 10^{? 4} and, 1 × 10^{? 5} mol L^{? 1}) resulted in a significant dose-dependent increase in the number of micronuclei in comparison with control (74.00 ± 13.00/1000 binucleated cells and 52.67 ± 10.12/1000 binucleated cells versus 30.33 ± 2. 82/1000 binucleated cells in DMSO control, P = 0.005 and 0.02, respectively, ANOVA followed by Tukey test P < 0.05). Many of the treated cells also possessed multiple micronuclei. Tolylfluanid did not affect the nuclear division index at all treatment concentrations. Our in vitro results thus demonstrate that tolylfluanid had a significant genotoxic effect at only the highest concentration tested.     

Evaluation of protective effects of sulforaphane on DNA damage caused by exposure to low levels of pesticide mixture using comet assay

The objective of the study was to evaluate the potential risk of DNA damage due to exposure to a mixture of the most widely used pesticides, namely endosulfan, chlorpyriphos and thiram at an environmentally relevant concentration (5 μM each) and the DNA protective capacity of sulforaphane (SFN) (10–30 μg/mL). DNA damage in human lymphocytes was ascertained with Single Cell Gel Electrophoresis (SCGE), also called Comet Assay. For positive control, H₂O₂ at 100 mM was used. The pesticide mixture produced DNA damage at the concentration used in the lymphocytes. SFN was able to offer a statistically significant (P < 0.01), concentration-dependant protection to DNA damage between 10–20 μg/mL in both the pre-incubation and co-incubation strategies. The results indicate that exposure to low levels of these pesticide mixtures can induce DNA damage, and the presence of SFN in diet may reduce the incidence of genetic damage, especially in farm workers. However, it is not clear whether SFN is involved in quenching of the free radicals generated by the pesticide mixture or it is involved in DNA repair mechanism.     

松胞素B对人血淋巴细胞和CHL细胞微核率的影响

研究了不同浓度的松胞素Ｂ对人血淋巴细胞和中国仓鼠肺成纤维细胞（ＣＨＬ）分裂吉周期和微核率的影响。结果表明,松胞素Ｂ浓度增高过高能引起细胞微核率增高。在本研究中,对ＣＨＬ细胞微核率试验以３μｇ／ｍＬ松胞素Ｂ较适合;而对人血淋巴细胞微核试验,松胞素Ｂ浓度以２－４μｇ／ｍＬ为宜,松胞表Ｂ浓度过高会提高微核背景值,降低试验的灵敏度和精确性。     

Genotoxic effects of metals on human salivary gland tissue and lymphocytes as detected by the Comet assay

The etiology of salivary gland malignancies still remains unclear. Metal compounds are of special interest since they show ubiquitous presence in the environment, are present in many working places, and are accepted (co-)carcinogens in some other malignancies. Metals enter the body as xenobiotics by inhalation or ingestion. This study investigated the genotoxic potential of sodium dichromate (Na₂Cr₂O₇), nickel sulfate (NiSO₄), cadmium sulfate (CdSO₄) and zinc chloride (ZnCl₂) on human salivary gland cells and lymphocytes. Macroscopically healthy tissue of salivary glands was harvested from 46 patients during surgery and isolated to single cells by enzymatic digestion. The cells were incubated with Na₂Cr₂O₇, NiSO₄, CdSO₄ or ZnCl₂. Na₂Cr₂O₇ was also incubated in combination with the other metal compounds listed. Carcinogenic and co-carcinogenic effects of cadmium were tested by incubation with Na₂Cr₂O₇ and consecutive repair intervals. DNA damage and repair were evaluated by the Comet assay, determining DNA-strand breaks. The extent of damage was quantified using a digital analysis system. Na₂Cr₂O₇ produced significantly enhanced DNA-strand breaks in human salivary gland tissue and lymphocytes. All other metal compounds exerted no damaging effect on both cell types. Co-incubation of Na₂Cr₂O₇ with the other metals revealed a significant additive effect only for CdSO₄. Specific analysis of the influence of cadmium showed a reduction of DNA-repair after Na₂Cr₂O₇-induced strand breaks in salivary gland cells. This study provides evidence that exposure to distinct metals may significantly contribute to malignant salivary gland tumors. In consequence, further studies as epidemiological and toxicological data are warranted to determine the role of distinct metals as potential (co-) carcinogens.     

二氧化硫对人体血淋巴细胞的遗传毒理效应

本文对某硫酸厂接触二氧化硫（ＳＯ＿２）４０名工人的外周血淋巴细胞染色体畸变（ＣＡ）、姊妹染色单体互换（ＳＣＥ）、及微核（ＭＮ）进行了分析。结果表明，接触ＳＯ＿２的工人其ＣＡ、ＳＣＥ及ＭＮ均显著高于对照组。本文也分析了ＳＯ＿２在体内的代谢产物──亚硫酸氢钠对淋巴细胞的遗传毒理效应。结果指出，亚硫酸氢钠可诱发人血淋巴细胞ＣＡ，ＳＣＥ及ＭＮ，且有明确的剂量效应关系。这些研究证明，ＳＯ＿２确是人血淋巴细胞染色体断裂剂和基因毒性因子；ＳＯ＿２与癌症的关系值得进一步研究。     

Genotoxic biomonitoring of agricultural workers exposed to pesticides in the north of Sinaloa State, Mexico

Genotoxic damage was evaluated in 70 agricultural workers, 25 women and 45 men, exposed to pesticides in Las Grullas, Ahome, Sinaloa, Mexico, with an average of 7 years of exposure. The effect was detected through the sister chromatid exchanges (SCE) in lymphocytes of peripheral blood and micronuclei (MN) and other nuclear anomalies (NA) in buccal exfoliated cells. Also, the influence on cellular proliferation kinetics (CPK) was studied by means of the replication index (RI) and the cytotoxic effect was examined with the mitotic index (MI). The non-exposed group consisted of 70 other persons, 21 women and 47 men from the city of Los Mochis, Sinaloa, Mexico. Significant differences between the exposed and the non-exposed groups were observed in SCE, CPK, MI, MN and NA. Analysis of variance revealed that age, gender, smoking and alcohol consumption did not have a significant effect on genetic damage. However, there was a correlation between exposure time to pesticides and SCE frequency. These results could have been due to the exposure of workers to pesticides containing different chemical compounds. This study afforded valuable data to estimate the possible risk to health associated with pesticide exposure.     

The effect of thiacloprid formulation on DNA/chromosome damage and changes in GST activity in bovine peripheral lymphocytes

The potential genotoxic effect of thiacloprid formulation on bovine peripheral lymphocytes was evaluated using the comet assay and the cytogenetic endpoints: chromosome aberrations (CAs), sister chromatid exchanges (SCEs) and micronuclei (MNi). Whole blood cultures were treated with the insecticide at concentrations of 30, 60, 120, 240 and 480 μg mL^?1 for 24, 48 h and/or 2 h of incubation. A statistically significant increase in the frequency of DNA damage, as well as in unstable chromosome aberrations (% breaks) were found after exposure to the insecticide at concentrations ranging from 120 to 480 μg mL^?1 (P < 0.05, P < 0.01, P < 0.001). For the detection of stable structural chromosome aberrations (e.g., translocations) and numerical aberrations by the FISH method, three whole chromosome painting probes for bovine chromosomes 1, 5 and 7 (BTA1, BTA5 and BTA7) were used in our experiments. We observed numerical aberrations, but without any statistical significance. Regarding the sister chromatid exchanges, no significant elevation in the SCE frequencies was found after 24-h exposure to the insecticide. A dose-related response in the SCE induction was obtained in bovine cultures after the prolonged time of exposure (48 h) to thiacloprid formulation at concentrations ranging from 120 to 480 μg mL^?1 in each donor (P < 0.05, P < 0.01), which was associated with a reduction of the PI (P < 0.05, P < 0.01). The insecticide failed to produce MNi; however, a significant reduction of CBPI was observed. Using real-time PCR, a decrease in the expression of bovine glutathione S-transferase M3 (GSTM3) was detected at the lowest dose. The higher concentrations of thiacloprid formulation caused an increase in the mRNA expression.     

Use of sensitive methods for detection of DNA damage on human lymphocytes exposed to p,p′-DDT: Comet assay and new criteria for scoring micronucleus test

Wide distribution, stability and long persistence in the environment of dichlorodiphenyltrichloroethane (DDT), probably the best-known and most useful insecticide in the world, imposes the need for further examination of the effect of this chemical on human health and especially on the human genome. In this study, peripheral blood human lymphocytes from a healthy donor were exposed to 0.025 mg/L concentration of p,p′-DDT at different time periods (1, 2, 24 and 48 h). For the assessment of genotoxic effect, the new criteria for scoring micronucleus test and alkaline comet assay were used. Both methods showed that p,p′-DDT induces DNA damage in low concentration used in this research. Results of micronucleus test showed a statistically significant (p < 0.05) genotoxic effect of p,p′-DDT on human lymphocytes compared with corresponding control and a different exposure time. A comet assay also showed increased DNA damage caused in p,p′-DDT-exposed human lymphocytes than in corresponding control cells for the tail length. Results obtained by measuring the level of DNA migration and incidence of micronuclei (MN), nucleoplasmic bridges (NPBs) and nuclear buds (NBUDs) indicate the sensitivity of these tests and their application in detection of primary genome damage after long-term exposure to establish the effect of p,p′-DDT on human genome.