ABSTRACT
【Objective】 To compare PC-12 cells’ apoptosis caused by the serotonin herbicides atrazine （ATR）， simazine （SIM） and cyanazine （CYA）. 【Methods】 The rat adrenal medullary pheochromoma PC-12 cell line was selected for routine culture. At the cell logarithmic growth phase， ATR， SIM and CYA were used at a concentration of 200 μmol/L for 24 h， respectively， and the same solvent was added in the control group. The CCK-8 method was used to detect the cell survival rate； the content of reactive oxygen species （ROS） in PC-12 cells was detected； the Real-time PCR and Western blotting methods were used to detect the mRNA and protein expressions of Bax， p53， Bcl-2 and Caspase-3. 【Results】 Compared with that in the control group， the survival rate of the cells in ATR group， SIM group and CYA group was significantly decreased. The intracellular ROS activity of the three groups was significantly increased， and the mRNA and protein expressions of Bax， p53 and Caspase-3 were increased. mRNA and protein expressions of Bcl-2 were significantly reduced （P<0.01）. Compared with that in ATR group， the cell survival rate of SIM group and CYA group was significantly increased， the intracellular ROS activity of the two groups was significantly decreased， the expressions of Bax， p53 and Caspase-3 mRNA and protein were significantly reduced， and the expressions of Bcl-2 mRNA and protein were significantly increased （P<0.01）. Compared with that of SIM group， the cell survival rate of CYA group was significantly increased， while the intracellular ROS activity was significantly decreased； the Bax， p53 and Caspase-3 mRNA and protein expressions were significantly reduced， and Bcl-2 mRNA and protein expressions were significantly increased （P<0.01）. 【Conclusion】 ATR， SIM and CYA can all promote PC-12 cells’ apoptosis； ATR has the strongest effect while CYA has the weakest effect.
ABSTRACT
Aim: To identify the oxidative stress impacts of chloro-s-triazine herbicides on human mammary epithelial cell lines.Study Design:MCF-7 mammary epithelial carcinoma and MCF-10A mammary epithelial cells were treated with levels of three triazine herbicides in concentrations flanking the US FDA safe levels. Place and Duration of Study:Department of Biology, Millikin University, in January 2015 through December 2015 and January 2019 through May 2020.Methodology:We examined the oxidative effects of two triazine herbicides, atrazine and simazine, on estrogen-dependent MCF-7 mammary epithelial carcinoma cells using three different bioluminescent assay techniques. We then utilized real time PCR to analyze gene expression through RT-PCR analysis, in both MCF-7 cells and a non-cancerous cell line, MCF-10A, for both of these triazine herbicides plus the related cyanazine.Results:At all concentrations of atrazine and simazine, no statistical differences were found in the levels of oxidized glutathione or total oxidized and reduced nicotinamide adenine dinucleotides phosphates. In stark contrast, levels of hydrogen peroxide were found tobe statistically different from the control at all concentrations of atrazine and simazine tested. Using an Analysis of Variance (ANOVA) we determined that within the enzymatic portion of the hydrogen peroxide pathway there were statistically significant differences in the expression of Peroxiredoxin 1 (PRDX1), Sulfiredoxin (SRXN1), and Thioredoxin (TXN).Conclusion:Exposure to triazines alters the hydrogen peroxide pathway, which in turn can greatly affect the stability of the cell milieu
ABSTRACT
A biorremediação é uma tecnologia que utiliza o metabolismo de microrganismos para eliminação ou redução, a níveis aceitáveis, de poluentes presentes no ambiente. Os herbicidas triazínicos são usados intensivamente no controle de ervas daninhas, principalmente na cultura de milho. Objetivou-se, neste trabalho, isolar fungos filamentosos de solos contaminados com herbicidas triazínicos (atrazine e simazine) e selecionar os microrganismos isolados quanto à capacidade de crescimento em meio adicionado de atrazine. Os microrganismos foram isolados, cultivados em meio Ágar-Batata-Dextrose (BDA) acidificado com ácido tartárico 10 por cento, adicionado de 50 mg.Kg-1 de atrazine e incubados por 5 dias a 25ºC. Foi realizada a medida diária do crescimento fúngico e calculada a velocidade de crescimento radial através de regressão linear dos raios das colônias utilizando-se a equação r(t) = a + VCR .t (r:raio; t: tempo; VCR: velocidade de crescimento radial). Os resultados de VCR foram analisados através de Anova simples e do teste de Tukey, para comparação de médias. Foram isolados 15 fungos, pertencentes aos gêneros Aspergillus, Penicillium e Trichoderma. As maiores VCRs foram obtidas com fungos Aspergillus (A1) e Penicillium (AS1), isolados de solo contaminado com atrazine e atrazine adicionado de simazine, respectivamente, que apresentaram VCRs de 1,57 mm.d-1 e 1,28 mm.d-1. O crescimento dos fungos em meio contaminado com a atrazine indica a possibilidade de utilização desses fungos em estudos de biorremediação de solos contaminados com herbicidas triazínicos.
Bioremediation is a technology that uses microrganism metabolism to quickly eliminate or reduce pollutants to acceptable levels into the environment. The triazine herbicides are intensively used to control harmful grass in the culture of maize. The aim of this work was to isolate filamentous fungi from soil contaminated with triazine herbicides and screening these fungi due to their ability of growth in a medium added by atrazine. The fungi were isolated, cultivated in potato-dextrose-agar plus 50 mg.Kg-1 of atrazine and incubated for 5 days at 25ºC. The measure of the rays of the colonies was carried out daily and the radial growth rate (RGR) through linear regression of colonies rays using the equation: r(t) = a + RGR .t (r:ray; t: time; RGR: radial growth rate). The RGR results were analyzed through analysis of variance and Tukey test for comparison of averages. Fifteen filamentous fungi from genus Aspergillus, Penicillium and Trichoderma were isolated. The highest RGRs were obtained with the fungi Aspergillus and Penicillium, when isolated from contaminated soil with atrazine and atrazine + simazine, respectively, showing RGR of 1.57 mm.d-1 and 1.28 mm.d-1. The growth of these fungi in atrazine contaminated meas indicates a possible of use of them in bioremediation experiments with contaminated soil containing triazine herbicides.