Department of Physical Chemistry, Vinca Institute of Nuclear Sciences, Belgrade, Serbia.
Toxicol Lett. 2010 Mar 1;193(1):9-18. doi: 10.1016/j.toxlet.2009.11.022. Epub 2009 Dec 3.
The toxic effects of diazinon and its irradiated solutions were investigated using cultivated human blood cells (lymphocytes and erythrocytes) and skin fibroblasts. Ultra Performance Liquid Chromatography (UPLC)-UV/VIS system was used to monitor the disappearance of starting diazinon during 115-min photodegradation and formation of its by-products (diazoxon and 2-isopropyl-6-methyl-4-pyrimidinol (IMP)) as a function of time. Dose-dependent AChE and Na(+)/K(+)-ATPase inhibition by diazinon was obtained for all investigated cells. Calculated IC(50) (72 h) values, in M, were: 7.5x10(-6)/3.4x10(-5), 8.7x10(-5)/6.6x10(-5), and 3.0x10(-5)/4.6x10(-5) for fibroblast, erythrocyte and lymphocyte AChE/Na(+)/K(+)-ATPase, respectively. Results obtained for reference commercially purified target enzymes indicate similar sensitivity of AChE towards diazinon (IC(50) (20 min)-7.8x10(-5)M), while diazinon concentrations below 10mM did not noticeably affect Na(+)/K(+)-ATPase activity. Besides, diazinon and IMP induced increasing incidence of micronuclei (via clastogenic mode of action) in a dose-dependent manner up to 2x10(-6)M and significant inhibition of cell proliferation and increased level of malondialdehyde at all investigated concentrations. Although after 15-min diazinon irradiation formed products do not affect purified commercial enzymes activities, inhibitory effect of irradiated solutions on cell enzymes increased as a function of time exposure to UV light and resulted in significant reduction of AChE (up to 28-45%) and Na(+)/K(+)-ATPase (up to 35-40%) at the end of irradiation period. Moreover, photodegradation treatment strengthened prooxidative properties of diazinon as well as its potency to induce cytogenetic damage.
采用超高效液相色谱-紫外可见分光光度法（UPLC-UV/VIS）系统监测了 115 分钟光解过程中起始敌百虫的消失以及其副产物（敌敌畏和 2-异丙基-6-甲基-4-嘧啶醇（IMP））的形成情况。研究了敌百虫及其辐照溶液对培养的人血细胞（淋巴细胞和红细胞）和成纤维细胞的毒性作用。结果表明，辐照溶液对所有被研究的细胞中的 AChE 和 Na(+)/K(+)-ATPase 均有剂量依赖性抑制作用。计算得到的 IC(50)（72 h）值（M）分别为：7.5x10(-6)/3.4x10(-5)、8.7x10(-5)/6.6x10(-5)和 3.0x10(-5)/4.6x10(-5)，分别为成纤维细胞、红细胞和淋巴细胞 AChE/Na(+)/K(+)-ATPase 的 IC(50)。对于参考的商业纯化靶酶，得到的结果表明 AChE 对敌百虫的敏感性相似（IC(50)（20 min）-7.8x10(-5)M），而低于 10mM 的敌百虫浓度不会明显影响 Na(+)/K(+)-ATPase 活性。此外，敌百虫和 IMP 以剂量依赖的方式诱导微核形成的发生率增加（通过断裂剂作用模式），直至 2x10(-6)M，并显著抑制细胞增殖和增加所有研究浓度下的丙二醛水平。虽然 15 分钟敌百虫辐照后形成的产物不会影响商业纯化酶的活性，但辐照溶液对细胞酶的抑制作用随暴露于紫外光的时间而增加，导致 AChE（高达 28-45%）和 Na(+)/K(+)-ATPase（高达 35-40%）的活性在辐照结束时显著降低。此外，光降解处理增强了敌百虫的促氧化性质及其诱导细胞遗传毒性的能力。
