Dichloroacetate- and trichloroacetate-induced modulation of superoxide dismutase,catalase, and glutathione peroxidase activities and glutathione level in the livers of mice after subacute and subchronic exposures

Dichloroacetate (DCA) and trichloroacetate (TCA) were previously found to induce various levels of oxidative stress in the hepatic tissues of mice after subacute and subchronic exposures. The cells are known to have several protective mechanisms against production of oxidative stress by different xenobiotics. To assess the roles of the antioxidant enzymes and glutathione (GSH) in DCA- and TCA-induced oxidative stress, groups of B6C3F1 mice were administered either DCA or TCA at doses of 7.7, 77, 154, and 410 mg kg^?1 day^?1, by gavage for 4 weeks (4-W) and 13 weeks (13-W), and superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) activities, as well as GSH were determined in the hepatic tissues. DCA at doses ranging between 7.7–410, and 7.7–77 mg kg^?1 day^?1, given for 4-W and 13-W, respectively, resulted in either suppression or no change in SOD, CAT, and GSH-Px activities, but doses of 154–410 mg DCA kg^?1 day^?1 administered for 13-W were found to result in a significant induction of the three enzyme activities. TCA administration on the other hand, resulted in increases in the SOD and CAT activities, but caused suppression of GSH-Px activity in both the periods. Except for the DCA doses of 77–154 mg kg^?1 day^?1 administered for 13-W that resulted in a significant reduction in the GSH levels, all other DCA as well as TCA treatments produced no changes in GSH. Since these enzymes are involved in the detoxification of the reactive oxygen species (ROS), superoxide anion (SA), and H₂O₂, it is concluded that SA is the main contributor to DCA-induced oxidative stress, while both ROS contribute to that of TCA. The increase in the enzyme activities associated with 154–410 mg DCA kg^1? day^?1 in the 13-W period suggest their role as protective mechanisms contributing to the survival of cells modified in response to those treatments.     

Humic substances. Part III. sorptive interactions with environmental chemicals†

The formation of covalent binding to DNA of a carcinogen is now widely accepted to represent a classical mechanism of tumour induction in mammals. This mechanism does not operate with metals since no covalent binding of these agents to DNA does occur. Nevertheless, somatic mutations as typical consequences of DNA‐damage have been reported to be induced by metals in various model systems. Beside DNA‐alkylation such damages can be caused by changes in the conformation of DNA or in the fidelity of DNA‐repair. The activity of the repair enzymes DNA‐polymerases is indeed impaired by many metal ions at least in vitro. It is not yet established whether these mechanisms are also important in the intact mammalian organism. Much evidence has accumulated during the last years that a disturbance of the balance of cations and especially metal ions represents another possible mechanism of tumour induction. The tumours found with high doses of chelating agents such as nitrilo‐triacetic acid (NTA) have to be discussed in this context. Since most—if not—all of the speculative mechanisms of metal carcinogenesis resemble classical pharmacological reactions the existence of a threshold level is likely. So metal carcinogenesis will not be a problem of the environmental contamination at trace levels but a problem of occupational medicine.     

文蛤谷胱甘肽抗氧化系统对北部湾沉积物中可提取态重金属的响应

为识别海洋双壳类谷胱甘肽抗氧化系统对滨海沉积物重金属可提取态和总量的响应特征差异,分析了北部湾潮间带沉积物中不同形态Cu、Pb、Zn、Cd、Cr的质量分数,文蛤(Meretrix meretrix)鳃组织中w(GSH)(GSH为还原型谷胱甘肽)、w(GSSG)(GSSG为氧化型谷胱甘肽)、GPx(谷胱甘肽过氧化物酶)和GST(谷胱甘肽硫转移酶)活性,并计算w(GSH)/w(GSSG)、tGSH(总谷胱甘肽)含量和OSI(氧化逆境指数). 结果表明:w(TE-Pb)(TE-Pb为可提取态Pb,下同)、w(TE-Cr)、w(TE-Cu)、w(TE-Zn)和w(TE-Cd)平均值分别为13.40、1.93、13.19、30.09和0.10mg/kg; w(TE-Cd)占w(T-Cd)(T-Cd为总Cd,下同)的60.2%~98.1%,并且主要为弱酸溶解态和有机物及硫化物结合态;部分采样点w(TE-Pb)和w(TE-Zn)较高,二者占w(T-Pb)和w(T-Zn)的平均值分别为44.6%和56.6%;Cr、Cu分别在全部或大多数采样点以残渣态为主,可提取态含量平均仅占总量的17.9%和36.5%. 重金属的可提取态质量分数与总量的空间分布基本一致(Cr除外). 文蛤鳃中w(GSH)、w(GSSG)分别为4.07~6.06、4.01~6.59mg/g. 抗氧化指标对沉积物中重金属总量和可提取态质量分数均有显著响应的为w(GSSG)与Cd,w(GSH)/w(GSSG)与Cr和Cd,OSI与Cr和Cd. 然而,w(GSH)/w(GSSG)和OSI只对w(T-Zn)有良好响应;GST只与w(TE-Cu)、w(TE-Zn)显著相关. 某些抗氧化指标对重金属总量和可提取态质量分数表现出不同程度的响应,因此,不能单独依据双壳类对重金属总量的生化响应特征来筛选用于沉积物综合毒性评价的生物标志物.      

亚致死剂量溴氰菊酯对麦穗鱼谷胱甘肽S-转移酶活性的影响

本文研究了麦穗鱼Pseudorasbora parva谷胱甘肽S-转移酶的组织分布以及亚致死剂量的溴氰菊酯对其比活力和组织分布的影响.结果表明肝胰脏、肠、鳃、卵巢、肾、脾脏、鳔、肌肉谷胱甘肽S-转移酶(GSTs)活性分别占41.73%、14.31%、12.11%、10.10%、8.62%、5.23%、4.33%和3.58%.用溴氰菊酯亚致死剂量活体处理麦穗鱼120 h后,卵巢和肾GSTs的活性与对照组具有显著差异,活性提高1.85倍;肠和鳃的GSTs活性分别是对照组的1.40倍和1.44倍;对肝脏GSTs活性的诱导不显著.这表明溴氰菊酯对麦穗鱼不同组织部位诱导的时间效应和强度是不同的.     

基于谷胱甘肽结合作用定量研究微囊藻毒素的解毒代谢机制

基于谷胱甘肽(GSH)解毒作用探讨了微囊藻毒素-RR(MCRR)在不同动物肝脏和肾脏合作下的代谢机制。通过人工合成MCRR的谷胱甘肽代谢物(MCRR-GSH),腹腔注射至鲫鱼和大鼠体内,利用液相色谱串联质谱技术(LC-MS/MS)定量检测MCRR-GSH及其下游半胱氨酸代谢物(MCRR-Cys)在组织内的代谢动力学变化。在72 h的暴露实验中,实验组鲫鱼和大鼠体内均定量检测到MCRR-GSH和MCRR-Cys。MCRR-GSH在肾脏中的浓度显著高于其他组织(P0.05),鲫鱼和大鼠体内累积浓度分别是(0.161±0.001)和(0.116±0.005)μg·g~(-1)DW。同样的,MCRR-Cys主要分布于鲫鱼和大鼠的肾脏组织。鲫鱼肾脏中MCRR-Cys的浓度出现明显的波动,而肝脏和胆汁内的MCRR-Cys浓度却呈现出上升的趋势;大鼠肾脏内MCRR-Cys的浓度呈缓慢下降的趋势,浓度范围为(8.899±0.817)μg·g~(-1)DW至(3.336±0.263)μg·g~(-1)DW。基于以上结果推测,微囊藻毒素在肝脏和肾脏合作下的解毒过程为:MC在肝脏内经GSH结合作用生成的代谢物MC-GSH随血液循环转运至肾脏,在肾脏内MCGSH快速地转化为下游代谢物MC-Cys以促进排泄。     

沉水植物对阿特拉津胁迫的毒理响应

为揭示在阿特拉津胁迫下沉水植物生长及其与谷胱甘肽代谢途径的关系,通过培养实验研究了沉水植物菹草(Potamogeton crispus)和穗花狐尾藻(Myriophyllum spicatum)对0、0.5、1.0和2.0 mg·kg~(-1)阿特拉津的吸收特性,并对不同培养时期沉水植物的鲜重、总谷胱甘肽含量(T-GSH,即还原型谷胱甘肽GSH和氧化型谷胱甘肽GSSG之和)、GSH/GSSG比值及其形态变化、谷胱甘肽还原酶(GR)活性和谷胱甘肽-S-转移酶(GST)活性进行了测定。结果表明:沉积物阿特拉津初始浓度越高,植物体内阿特拉津浓度也越高。在培养60 d内,添加的阿特拉津对2种沉水植物的生长均产生显著抑制作用(P0.05)。在阿特拉津胁迫60 d后,各处理植物体内GSH/GSSG比值有所回升,其GR和GST活性均高于空白对照组。处理组植物体内GR和GST活性在30 d时达到最高值。与此同时,GSH脱去谷氨酸后能与阿特拉津形成共轭物。以上结果提示,≤2 mg·kg~(-1)阿特拉津在培养前60 d内会对植物生长产生抑制,但在90 d时植物会从伤害中恢复过来。沉水植物体内的谷胱甘肽在GR和GST作用下,对阿特拉津及其产生的活性氧具有一定去除作用,并通过控制酶活使植物体保持一定的GSH含量;另一方面,GSH可以与阿特拉津结合形成新的共轭物,以此缓解阿特拉津对沉水植物的毒害。     

增塑剂邻苯二甲酸二异壬酯对雄性小鼠生殖毒性的氧化损伤机制

研究增塑剂邻苯二甲酸二异壬酯(diisononyl phthalate,DINP)对雄性小鼠生殖毒性的氧化损伤机制,以昆明雄性小鼠为受试动物,随机分为7组,包括空白对照组(生理盐水),4个DINP暴露组(0.2、2、20和200 mg·kg~(-1))和褪黑素(melatonin,Mel)对照组50 mg·kg~(-1)、Mel处理组(200 mg·kg~(-1)DINP+50 mg·kg~(-1)Mel),灌胃14 d。以睾丸组织匀浆测定活性氧(ROS)、还原型谷胱甘肽(GSH)和8-羟基脱氧鸟苷(8-OHdG)含量;以睾丸组织细胞测定DNA-蛋白交联(DPC)系数。随着DINP暴露剂量的增加,DINP暴露组睾丸组织ROS、8-OHdG含量和DPC系数逐渐上升,GSH含量逐渐降低,差异有统计学意义(P0.05,P0.01); Mel处理组ROS、8-OHdG含量和DPC系数相应降低,GSH含量逐渐上升。雄性小鼠睾丸组织形态观察结果表明,随着DINP暴露剂量的增加,小鼠睾丸组织的病理损伤程度呈上升趋势。实验结果表明,较高剂量(≥20 mg·kg~(-1))的DINP能造成小鼠睾丸组织的病理损伤和氧化损伤,50 mg·kg~(-1)Mel的抗氧化能力可以有效对小鼠睾丸组织起保护作用,使组织损伤减轻,即生殖毒性减弱。     

环境激素甲草胺对摇蚊谷胱甘肽硫转移酶的抑制作用

谷胱甘肽硫转移酶(GST)是生物体内重要的解毒酶。为研究环境激素甲草胺对溪流摇蚊GST的抑制作用及机理,采用酶标仪微量法测定甲草胺对溪流摇蚊4龄幼虫GST的抑制活性及抑制类型。结果表明甲草胺在72 h活体染毒和离体状态下均能显著抑制溪流摇蚊GST的活性,且抑制程度随着药物浓度的增大而增大。甲草胺抑制GST活性的反应为可逆性抑制。进一步的酶动力学分析揭示甲草胺对GST的特异底物1-氯-2,4-二硝基苯(CDNB)和还原型谷胱甘肽(GSH)均表现为竞争性抑制,即甲草胺仅影响米氏常数(Km),不改变酶促反应的最大反应速度(Vmax)。以上结果表明农药残留污染物甲草胺在胁迫下对摇蚊幼虫GST酶活性会受到显著影响,甲草胺对GST的抑制机制为可逆性和竞争性抑制。     

邻苯二甲酸二异壬酯致小鼠肾组织氧化损伤的研究

邻苯二甲酸二异壬酯是一种新型替代增塑剂,对其毒性研究已受到广泛关注。为探究邻苯二甲酸二异壬酯对肾组织的氧化损伤作用,将昆明小鼠随机分为5组,包括1个阴性对照组、4个邻苯二甲酸二异壬酯染毒组,按0.5、5、50和500 mg·kg-1四个剂量水平灌胃染毒14天。制备小鼠肾组织切片进行病理学观察,以肾组织匀浆测定活性氧(reactive oxygen species,ROS)、还原型谷胱甘肽(glutathione,GSH)、丙二醛(malondialdehyde,MDA)和8-羟基脱氧鸟苷(8-hydroxy-2-deoxyguanosine,8-OHd G)的含量,以肾组织细胞悬液测定DNA-蛋白质交联(DNA-protein Crosslink,DPC)系数。随着邻苯二甲酸二异壬酯染毒剂量的升高,小鼠肾组织的损伤程度加重,ROS、MDA、8-OHd G含量和DPC系数逐渐上升,GSH含量逐渐降低,各指标均呈一定剂量-效应关系。染毒剂量为5 mg·kg-1时,ROS、DPC系数差异有统计学意义(P0.05,P0.01);染毒剂量为50 mg·kg-1和500 mg·kg-1时,ROS、GSH、8-OHd G、MDA含量和DPC系数差异均有统计学意义(P0.05,P0.01)。结果表明较高剂量(≥50 mg·kg-1)邻苯二甲酸二异壬酯可造成小鼠肾组织氧化损伤。     

Characterization and phylogenetic analysis of a phenanthrene-degrading strain isolated from oil-contaminated soil

Bacterium strain EVA17 was isolated from an oil-contaminated soil, and identified as Sphingomonas sp.based on analysis of 16S rDNA sequence, cellular fatty acid composition and physiological-chemical tests. The salicylate hydroxylase and catechol 2, 3-dioxygenase (C23O) were detected in cell-free lysates, suggesting a pathway for phenanthrene catabolism via salicylate and catechol. Alignment showed that both of the C23O and GST genes of the strain EVA17 had high similarity with homologues of strains from genus Sphingomonas. The phylogenetic analysis based on 16S rDNA and C23O gene sequence indicated that EVA17 should be classified into genus Sphingomonas, although the two phylogenetic trees were slightly different from each other. The results of co-amplification and sequence determination indicated that GST gene should be located upstream of the C230 gene.