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.     

Comparative Toxicity of PCBs and PBDEs Using Human Cancer Cell Lines and Zebrafish Embryos

研究了6种多氯联苯(PCBs)3,3′,4,4′-四氯联苯(PCB77)、2,3,3′,4,4′-五氯联苯(PCB105)、2,3′,4,4′,5-五氯联苯(PCB118)、3,3′,4,4′,5-五氯联苯(PCB126)、2,3,3',4,4',5-六氯联苯(PCB156)和商业型混合多氯联苯Aroclor1254,两种多溴联苯醚(PBDEs)2,2′,4,4′-四溴二苯醚(PBDE47)、十溴二苯醚(PBDE209)对人类癌细胞生长和斑马鱼脱膜与不脱膜胚胎发育的影响.8种化合物均使用0.01、0.1、1.0、10μmol·L-14个浓度进行1~6d的暴露实验.结果表明,PBDE209在最高浓度10μmol·L-1下对结肠癌细胞HCT116(暴露3d后)和RKO(暴露5d后)具有显著的生长抑制作用,所有化合物均对乳腺癌细胞没有显著影响.相比之下,化合物对受精后5~6h(5~6hpf)的斑马鱼胚胎的毒性效应显得比较明显,而各化合物对胚胎的致畸和致死效应又不相同,其毒性强弱依次为PCB126≈PCB156>PCB1254(Aroclor1254)>PBDE47>PCB77>PCB105≈PCB118≈PBDE209.其中PBDE209在未脱膜暴毒后均无致畸与致死现象,脱膜暴毒后最高浓度才表现出显著意义的致畸作用,而PBDE47在最高浓度下可产生高达80%的致畸率,这说明胚胎绒毛膜具有有效阻挡大分子物质如PBDE209进入的作用.PCBs的毒性效应与其空间结构密切相关.如PCB126和PCB105具有相同的分子式,前者在1μmol·L-1下就引起了显著的致死和致畸效应,而后者即使在10μmol·L-1下也没有显著的效应.实验结果也说明不同类型的实验对象所展示的毒性效应并不相同,化合物对体外培养的细胞和发育中的胚胎具有不同的影响.     

婴儿奶嘴提取液对大鼠肝原代细胞和HepG2细胞的毒性作用（Cytotoxicity of Extraction Liquids of Nipples on Primary Cultured Rat Hepatocytes and HepG2 Cells）

乳胶制品,如医用手套、婴儿奶嘴、避孕套等,存在着N-亚硝胺及亚硝基物质析出现象.对于乳胶制品中析出的N-亚硝胺及亚硝基物质,我国尚无法规加以限制,对其使用的安全性进行科学的分析更是鲜有报道.论文使用人工唾液盐溶液浸泡婴儿奶嘴,将提取液与体外培养的SD大鼠肝原代细胞、HepG2细胞作用一定时间,通过MTT实验和单细胞凝胶电泳实验,检测了婴儿奶嘴提取液对细胞存活率的影响和对DNA的损伤作用.结果显示:排除唾液盐溶液对细胞存活率的影响,婴儿奶嘴提取液与肝原代细胞和HepG2细胞作用24、48、72h,细胞存活率分别降至(85.22±13.77)%、(60.40±20.51)%、(79.28±20.08)%;(71.23±18.22)%、(69.51±16.96)%、(70.25±16.57)%,与阴性对照组(10%DEME)相比均有显著性差异(p<0.01).奶嘴提取液与肝原代细胞和HepG2细胞作用24h,尾长、Olive尾矩、尾DNA%、头尾比分别为(24.86±18.88)μm、6.60±5.99、(52.23±16.90)%、0.86±0.57;(17.48±10.87)μm、9.39±3.23、(57.94±20.94)%、0.83±0.67,显著高于阴性对照组和唾液盐溶液组(p<0.01).研究结果表明婴儿奶嘴提取液能够降低肝原代细胞和HepG2细胞存活率,且对DNA具有损伤作用.     

四溴二苯醚对人甲状腺细胞功能的影响

为研究四溴二苯醚(BDE-47)对原代培养人甲状腺细胞功能的影响,分别用浓度为10-12、10-10和10-8mol·L-1的BDE-47处理原代人甲状腺滤泡上皮细胞(TEC)24h,采用化学发光酶联免疫检测法检测细胞上清液中甲状腺球蛋白(Tg)的浓度,半定量逆转录聚合酶链反应技术检测功能相关的Tg基因和双链复合蛋白8(Pax-8)基因的表达.结果表明,10-12、10-10mol·L-1暴露组Tg分泌量较对照组显著减少(p<0.05),而10-8mol·L-1BDE-47组Tg分泌量与对照组无显著性差异(p>0.05).甲状腺功能相关Tg基因和Pax-8基因的表达量随BDE-47浓度的增加而显著降低(p<0.05),呈明显的剂量-效应关系.以上结果提示,BDE-47对离体培养的原代人甲状腺细胞的功能具有抑制作用,甲状腺功能相关Tg基因和Pax-8基因的表达下调可能是其作用机制之一.     

铅暴露U251细胞差异表达基因及相关通路(The Interrelated Pathways in Brain Human U251 Glioma Cells by Lead Exposure)

为探讨铅对体外培养的人神经胶质瘤U251细胞(human U251glioma cells,U251)暴露后基因表达的变化以及相关基因通路,选用乙酸铅暴露U251细胞.细胞在乙酸铅中暴露8h和24h后提取RNA,使用cDNA芯片分析基因表达情况,芯片扫描结果经归一化处理,设定Ratio值<0.5或≥2.0为表达有差异基因.结果表明,铅暴露U251细胞导致2840条基因差异表达,使用KEGG和BioCarta数据库分析代表性基因网络.结果发现,铅暴露U251细胞导致大量基因差异表达,涉及多个代谢及信号通路,与神经组织相关的主要信号通路有Ca2+信号通路、Jak-STAT信号通路、MAPK信号通路、Wnt信号通路等,还涉及配体-受体、细胞因子相互作用等.这些通路相互联结,构成复杂的网络系统,调控细胞的生物学功能.     

四氯联苯对鸡外源原始生殖细胞迁移的影响

为了研究环境污染物四氯联苯对移植的原始生殖细胞增殖性迁移的影响,将发育至14期的供体鸡胚原始生殖细胞用PKH26荧光染料标记,然后移植到经过四氯联苯处理的受体鸡胚生殖新月区域,继续孵化,检测其迁移和增殖情况.结果表明,和对照组相比,四氯联苯可明显降低移植原始生殖细胞在血液和性腺中的数量;四氯联苯对外源性原始生殖细胞迁移有明显影响,但雌二醇对外源性原始生殖细胞的增殖性迁移无明显影响,揭示四氯联苯可能影响的是受体性腺的诱引物质,且这种影响是非雌激素样的作用.     

DNA-Protein Crosslinks Induced by Microcystin-LR in Hepatic Cells of Mice

为探讨微囊藻毒素-LR致小鼠肝细胞的DNA-蛋白质交联作用,将20只昆明雄性小鼠随机分为4组：1个对照组和3个染毒组,采用腹腔注射进行染毒7d,染毒剂量分别为3.0、6.0和12.0μg·kg-1,检测小鼠肝细胞DNA-蛋白质交联程度. 结果显示,3.0、6.0和12.0μg·kg-1微囊藻毒素-LR均可导致小鼠肝细胞显著的DNA-蛋白质交联作用（与对照组相比,p<0.01,p<0.01,p<0.05）,当微囊藻毒素为6.0μg·kg-1时,这种作用最明显.     

北京市大气细颗粒物的遗传和非遗传毒性研究

用胞质阻断微核试验与单细胞凝胶电泳法检测北京市大气PM_2.5无机提取物与有机提取物对Balb/c 3T3细胞微核形成与DNA链断裂的影响;通过划痕染料示踪技术(SL/DT)观察PM_2.5无机提取物与有机提取物对Balb/c3T3细胞间通讯的影响.发现PM_2.5有机提取物可引起双核微核细胞率显著增加(P<0.01)及导致慧星细胞率和DNA迁移长度显著增加(P<0.01);PM_2.5有机提取物引起细胞间通讯的抑制; PM_2.5无机提取物未见明显毒作用.结果表明,PM_2.5可引起染色体损伤和原发性DNA损伤,抑制细胞间通讯,其毒作用主要由其有机成分引起.     

钻井井场噪音对听觉系统影响的观察

对油田井场的特定部位进行声强测定及频谱分析,发现其噪声强度均已超过国家标准。选健康,耳廊反射正常豚鼠30只,分别置于柴油机旁（8只）,钻井平台（8只）和值班房（8只）,余6只为正常对照。每天暴露6h,连续21d,记录豚鼠脑干电位及扫描电镜下形态学改变,结果表明,虽然钻井平台上的噪音强度低于柴油机组,但它对人耳的损伤程度却高于柴油机组。     

Selective effects of fenitrothion on murine splenic T-lymphocyte populations and cytokine/granzyme production

The aim of this study was to investigate in vitro effects of fenitrothion (FNT) on mouse splenic lymphocytes. Here, naïve mice had their spleens harvested and splenocytes isolated. After exposure to FNT for 48 hr: splenocyte viability was measured using a tetrazolium dye assay; cell phenotypes, i.e., B-cells (CD19⁺), T-cells (CD3⁺), and T-cell subsets (CD4⁺ and CD8⁺), were quantified by flow cytometry; and, production of cytokines/granzyme-B was assessed via enzyme-linked immunosorbent assay. The ability for FNT to induce oxidative stress in the cells was evaluated by measuring hydroxyl radical (·OH) and malondialdehyde (MDA) production and changes in glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) activity. The results showed that FNT significantly inhibited splenocyte proliferation, and decreased production of interleukin (IL)-2, interferon gamma, IL-4, and granzyme B, but had no impact on IL-6 production. FNT also selectively decreased splenic T-cell levels but did not induce changes in CD19⁺ B-cells. Further, within the T-cell populations, percentages of CD3⁺, CD4⁺, and CD8⁺ T-cells (particularly CD8⁺ T-cells) were reduced. Lastly, FNT selectively increased MDA and ·OH production and inhibited SOD and GSH-Px activities in the splenic lymphocytes. These findings suggest that, due to oxidative damage, FNT selectively inhibits splenic T-lymphocyte survival and cytokine/granzyme production in vitro.