硫化物胁迫对日本沼虾呼吸代谢和能量代谢酶的影响

研究了硫化物暴露后日本沼虾细胞色素氧化酶(CCO)、琥珀酸脱氢酶(SDH)、延胡索酸还原酶(FRD)和乳酸脱氢酶(LDH)4种呼吸代谢酶及能量代谢酶-精氨酸激酶(AK)活性的变化规律.将日本沼虾暴露于0.6 mg·L~(-1)、2mg·L~(-1)的2个硫化物质量浓度组和不含硫化物的对照组水体中,暴露后0、2、12、24、48 h和解除暴露后48 h取肝胰腺和肌肉组织进行酶活性分析.结果显示:肝胰腺和肌肉组织SDH和CCO活性随硫化物质量浓度升高或暴露时间延长而显著降低(P<0.05).FRD、LDH和AK活性随硫化物质量浓度升高或暴露时间延长而显著升高(P<0.05).解除硫化物暴露后48 h,各质量浓度组酶活性与对照组无显著差异.上述酶活性还存在组织差异,肝胰腺呼吸代谢酶活性高于肌肉中相应酶活性,而AK活性与此相反.结果表明,硫化物胁迫导致日本沼虾有氧呼吸代谢减弱,无氧呼吸代谢增强,并动用其能量贮存物质磷酸精氨酸,产生更多ATP以适应外界不良环境.     

不同季节大气可吸入颗粒物(PM_(10))诱导小鼠原代培养神经元炎性及其级联损伤效应

为了探讨不同季节可吸入颗粒物(PM10)对神经元的损伤效应,本研究通过建立小鼠大脑皮层原代神经元体外染毒模型,考察了不同季节PM10对炎性因子诱导型一氧化氮合酶(iNOS)、环氧化酶-2(COX-2)和黏附分子(ICAM-1)表达水平的影响,并探讨了与之耦联的磷酸化Ca2+/钙调蛋白依赖性蛋白激酶Ⅱα(p-Ca MKⅡα)、环磷腺苷效应元件结合蛋白(p-CREB)及即早基因(c-jun、c-fos)的表达情况.结果表明,PM10暴露刺激神经元炎性细胞因子释放增加,显著上调p-Ca MKⅡα和p-CREB的水平,并刺激即早基因的高表达,且以冬季PM10的效应最为明显.由此提示,PM10暴露可能通过炎症反应机制激活Ca2+-Ca MK-CREB通路,进而刺激即早基因表达引发神经毒性作用,而冬季PM10中多环芳烃类物质负荷可能是造成效应季节性差异的主要原因.     

Tumor promoting effects of cyanobacterial extracts are potentiated by anthropogenic contaminants--evidence from in vitro study

Inhibition of gap junctional intercellular communication (GJIC) is affiliated with tumor promotion process and it has been employed as an in vitro biomarker for evaluation of tumor promoting effects of chemicals. In the present study we investigated combined effects of anthropogenic environmental contaminants 2,2′,4,4′,5,5′-hexachlorobiphenyl (PCB 153) and fluoranthene, cyanotoxins microcystin-LR and cylindrospermopsin, and extracts of laboratory cultures of cyanobacteria Aphanizomenon gracile and Cylindrospermopsis raciborskii, on GJIC in the rat liver epithelial cell line WB-F344. Binary mixtures of PCB 153 with fluoranthene and the mixtures of the two cyanobacterial strains elicited simple additive effects on GJIC after 30 min exposure, whereas microcystin-LR and cylindrospermopsin neither inhibited GJIC nor altered effects of PCB 153 or fluoranthene. However, synergistic effects were observed in the cells exposed to binary mixtures of anthropogenic contaminants (PCB 153 or fluoranthene) and cyanobacterial extracts. The synergistic effects were especially pronounced after prolonged (6-24 h) co-exposure to fluoranthene and A. gracile extract, when mixture caused nearly complete GJIC inhibition, while none of the individual components caused any downregulation of GJIC at the same concentration and exposure time. The effects of cyanobacterial extracts were independent of microcystin-LR or cylindrospermopsin, which were not detected in cyanobacterial biomass. It provides further evidence on the presence of unknown tumor promoting metabolites in cyanobacteria. Clear potentiation of the GJIC inhibition observed in the mixtures of two anthropogenic contaminants and cyanobacteria highlight the importance of combined toxic effects of chemicals in complex environmental mixtures.     

Modulation of the DNA repair system and ATR-p53 mediated apoptosis is relevant for tributyltininduced genotoxic effects in human hepatoma G2 cells

The toxic effects of tributyltin (TBT) have been extensively documented in several types of cells, but the molecular mechanisms related to the genotoxic effects of TBT have still not been fully elucidated. Our study showed that exposure of human hepatoma G2 cells to 1-4 μmol/L TBT for 3 hr caused severe DNA damage in a concentration-dependent manner. Moreover, the expression levels of key DNA damage sensor genes such as the replication factor C, proliferating cell nuclear antigen and poly (ADP-ribose) polymerase-1 were inhabited in a concentration-dependent manner. We further demonstrated that TBT induced cell apoptosis via the p53-mediated pathway, which was most likely activated by the ataxia telangiectasia mutated and rad-3 related (ATR) protein kinase. The results also showed that cytochrome c, caspase-3, caspase-8, caspase-9, and the B-cell lymphoma 2 were involved in this process. Taken together, we demonstrated for the first time that the inhibition of the DNA repair system might be more responsible for TBT-induced genotoxic effects in cells. Then the generated DNA damage induced by TBT initiated ATR-p53-mediated apoptosis.     

Selective cellular and molecular effects of beryllium on lymphocytes†

An involvement of the immune system has been implicated in many of the whole animal effects of beryllium (Be) and its compounds and thus lymphocytes are a relevant cell model to study some of the cellular and molecular actions of the metal. Recent results indicate micromolar concentrations of Be salts are weakly mitogenic to lymphocytes in vitro probably as a result of direct interactions with the cell membranes. Furthermore evidence has been obtained that high affinity lymphocyte Be‐binding may be predictive of the potential to express delayed Be hypersensitivity in vivo. Although sub mitogenic concentrations of Be salts may actually activate accessory (macrophage type) cells, concentrations normally mitogenic to lymphocytes were found to inhibit their proliferative response to accessory cell dependent mitogens as a result of a Be²⁺ concentration dependent destruction of the adherent cell population. Higher concentrations of Be salts proved cytostatic to lymphocytes possibly by the established property of intracellularly accumulated Be²⁺ to inhibit cell division. Lymphocyte proliferation can be related to increases in protein phosphoryl‐ation capacity which in vitro can be inhibited by Be²⁺ and preliminary evidence indicates that tyrosine phosphorylation may be particularly sensitive.     

非生物胁迫对盐生杜氏藻DsMPK转录、翻译和磷酸化修饰的影响

促有丝分裂活化蛋白激酶(Mitogen-activated protein kinase,MAPK或MPK)途径是在真核生物中广泛存在的调控多种生理过程的信号途径,在细胞增殖、分化、凋亡、生物和非生物胁迫等生理过程的调控中起着关键的作用.盐生杜氏藻(Dunaliella salina,盐藻)是目前世界上最耐盐的真核光合生物之一.在前期实验中,已发现盐藻MAPK(DsMPK)的表达受到低温和高盐胁迫的抑制.分析DsMPK在高温、低盐、氧化、紫外胁迫中的表达变化,发现DsMPK还受到氧化胁迫的抑制.进而分析DsMPK在高温、低盐、氧化、低温、高盐和紫外胁迫中翻译和磷酸化修饰的变化,发现除低盐胁迫外,在磷酸化修饰水平DsMPK同样受到各种非生物胁迫的抑制.在对不同生长阶段盐藻DsMPK磷酸化变化的分析中还发现,DsMPK的磷酸化水平与生长速率有一致性.各方面DsMPK的变化趋势说明其为一调控生长相关的MAPK.     

几种化合物对温石棉诱导人胚肺细胞蛋白激酶C活性的影响

为了探讨几种化合物对温石棉诱导人胚肺(HEL)细胞蛋白激酶C(PKC)活性的影响,该研究用不同浓度的柠檬酸铝、混合稀土或亚硒酸钠溶液浸泡温石棉1小时后,再将其与HEL细胞共同孵育24小时,测定了HEL细胞胞浆和胞膜中PKC的活性。其结果显示:随着石棉剂量的增加,HEL细胞胞浆和胞膜中的PKC活性逐渐升高,均呈明显的剂量依赖性;经3种化合物预处理的石棉,其所诱导的HEL细胞胞浆和胞膜中的PKC活性均明显低于未处理温石棉组。笔者提示:温石棉可能通过改变细胞内PKC的活性进一步促进肿瘤的发生;用3种化合物预处理温石棉,均可抑制温石棉对HEL细胞PKC活性的诱导作用。     

聚磷菌生物除磷机理研究进展

废水中过量磷酸盐是引起水体富营养化的主要原因之一,生物除磷是一项新型的水处理技术。聚磷菌生物在多聚磷酸盐的合成和降解过程中发挥重要作用。文章综述了目前聚磷菌生物除磷的生化机理,聚磷过程中涉及的主要酶类及磷酸盐转运过程相关基因的表达调控。     

转聚磷激酶基因的大肠杆菌去除水体中的磷

为探索有效的生物除磷方法,构建了聚磷激酶基因(ppk)的表达载体pET28a(+),并转化大肠杆菌(Escherichiacoli)BL21,获得了可过表达ppk基因的工程菌BL-PPK.RT-PCR结果表明,ppk基因在转化的大肠杆菌中得到了高效表达.聚磷试验结果表明,培养10h后,转化了ppk基因的大肠杆菌细胞内聚磷含量比对照菌株高20倍,而培养液中可溶性磷浓度为对照菌株的约1/9.