[Objective] This study aimed to investigate the effect of zearalenone (ZEN) on DNA damage of porcine leydig cells. [Method] Porcine leydig cells cultured in vitro were collected to determine the median lethal dose (LD...[Objective] This study aimed to investigate the effect of zearalenone (ZEN) on DNA damage of porcine leydig cells. [Method] Porcine leydig cells cultured in vitro were collected to determine the median lethal dose (LD50) of ZEN with tetrazolium-based colorimetric assay (MTT assay). Comet assay was carried out to detect the DNA damage of porcine leydig cells exposed to at 0 (negative group), 1, 5, 10, 20, 40 μmol/L of ZEN. [Result] The percentage of cell tail was 16.67%, 34.00%, 40.67%, 52.00% and 64.67% under 0, 1, 5, 10 and 20 μmol/L of ZEN, respectively; the differences between the percentages of cell tail in various experimental groups had extremely significant statistical significance compared with the negative group (P<0.01), showing a significant dose-effect relationship; Tail length in various groups was 57.60±4.78, 57.75±6.25, 78.97±5.83, 100.50±6.94 and 146.83±12.31 μm, respectively; Tail DNA % in various groups was 21.29±2.25%, 22.24±2.43%, 31.21±6.27%, 37.45±4.33% and 60.68±9.83%, respectively; Tail length and Tail DNA % in experimental groups with ZEN concentration above 5 μmol/L showed significant differences (P<0.05) compared with the negative group, which showed an upward trend with the increase of ZEN concentration. [Conclusion] ZEN has genotoxic effect on porcine leydig cells, which can cause DNA damage, with a significant dose-effect relationship.展开更多
Objective To assess the effect of benzene on sperm DNA damage ;Methods Twenty-seven benzene-exposed workers were selected as exposed group and 35 normal sperm donors as control group. Air concentration of benzene seri...Objective To assess the effect of benzene on sperm DNA damage ;Methods Twenty-seven benzene-exposed workers were selected as exposed group and 35 normal sperm donors as control group. Air concentration of benzene series in workshop was determined by gas chromatography. As an internal exposure dose of benzene, the concentration of trans, trans-muconic acid （ttMA） was determined by high performance liquid chromatography. DNA was detected by modified single cell gel electrophoresis （SCGE）. Results The air concentrations of benzene, toluene and xylene at the workplace were 86.49±2.83 mg/m^3, 97.20±3.52 mg/m^3 and 97.45± 2.10 mg/m^3, respectively. Urinary ttMA in exposed group （1.040 ± 0.617 mg/L） was significantly higher than that of control group （0.819 ± 0.157 mg/L）. The percentage of head DNA, determined by modified SCGE method, significantly decreased in the exposed group （n=13, 70.18% ± 7.36%） compared with the control （n=16, 90.62% ± 2.94%）（P〈0.001）. Conclusion The modified SCGE method can be used to investigate the damage of sperm DNA. As genotoxin and reprotoxins, benzene had direct effect on the germ cells during the spermatogenesiss.展开更多
[Objective] The like-rocket immunoelectrophoresis was used to explore a new feasible electrophoresis method for single cell gel electrophoresis assay （comet assay）.[Method] The like-rocket immunoelectrophoresis was ...[Objective] The like-rocket immunoelectrophoresis was used to explore a new feasible electrophoresis method for single cell gel electrophoresis assay （comet assay）.[Method] The like-rocket immunoelectrophoresis was used for single cell gel electrophoresis assay to detect DNA damage at single cell level,then it was compared with traditional electrophoresis method to analyze its advantage and disadvantages.[Result] Under cell DNA undamaged state,the results of two electrophoresis methods were consistent.When cell DNA was damaged,the comet tail divergence of some cells under traditional electrophoresis method were drifted,however,the comet tail image of like-rocket immunoelectrophoresis was concentrated and not shifted.[Conclusion] The like-rocket immunoelectrophoresis had some advantages.展开更多
DNA damage is one of the most important consequences of oxidative stress in the cells. If DNA repair is unable to modify these inducible DNA damages, genomic instability may lead to mutation, cancer, aging and many ot...DNA damage is one of the most important consequences of oxidative stress in the cells. If DNA repair is unable to modify these inducible DNA damages, genomic instability may lead to mutation, cancer, aging and many other diseases. Single cell gel electrophoresis or comet assay is a common and versatile method to quantify these types of DNA damages. DNA damages induced by hydrogen peroxide(H_2O_2) are one of the proper models for measurement of protective ability of different compounds. So the main aim of this review is to provide an overview about protection ability of medicinal plants and their potential mechanism against H_2O_2 induced DNA damages. In this review, relevant researches on the effect of medicinal plants on DNA damages induced by H_2O_2 and possible molecular mechanisms are discussed.It seems that, medicinal plants are considered as therapeutic key factors to protect DNA from consequences caused by oxidative stress. Sufficient in vitro evidences introduce them as DNA protective agents through different mechanisms including antioxidant activity and some other cellular mechanisms. Moreover, in order to correlate the antigenotoxicity effects with their potential antioxidant property, most of medicinal plants were evaluated in term of antioxidant activity using standard methods. This review highlights the preventive effects of herbal medicine against oxidative DNA damages as well as provides rational possibility to engage them in animal studies and future clinical investigations.展开更多
The effects of heavy metals Cd^(2+),Pb^(2+)and Zn^(2+)at 0.05,0.5 and 5.0 mg/L level and their interactions at 0.5 mg/L level on DNA damage in hepatopancreas of loach Misgurnus anguillicaudatus for 1-35 days exposure ...The effects of heavy metals Cd^(2+),Pb^(2+)and Zn^(2+)at 0.05,0.5 and 5.0 mg/L level and their interactions at 0.5 mg/L level on DNA damage in hepatopancreas of loach Misgurnus anguillicaudatus for 1-35 days exposure were examined by single cell gel electrophoresis(SCGE).For each test group,20 loaches with similar body size(5.17-7.99 g;11.79-13.21 cm)were selected and kept in aquaria with dechlori-nated water at(22±1)℃and fed a commercial diet every 48 h.According to the percentage of damaged DNA with tail and its TL/D(tail length to diameter of nucleus)value,the relationship between DNA damage degree and heavy metal dose and exposure time was determined.Results showed that the percentage of damaged DNA and the TL/D value were increased with the prolonged exposure time.The highest percentage(84.85%)of damaged DNA was shown in 5.0 mg/L Zn^(2+)group after 28 days exposure and the biggest TL/D value(2.50)in all treated groups after 35 days exposure.During the first treated week,the damnification of DNA was mainly recognized as the first level,after that time,the third damaged level was mostly observed and the percentage of damaged DNA was beyond 80%.The joint toxic effects among Cd^(2+),Pb^(2+)or Zn^(2+)revealed much complexity,but it generally displayed that the presence of Cd^(2+)could enhance the genotoxicity of Pb^(2+)or Zn^(2+).In conclusion,the results suggestedthattherewasasignificanttime-anddose-depended relationship between the heavy metal and DNA damage in hepatopancreas of loach,and SCGE could represent a useful means to evaluate the genotoxicity of environmental contamination on aquatic organisms.展开更多
文摘[Objective] This study aimed to investigate the effect of zearalenone (ZEN) on DNA damage of porcine leydig cells. [Method] Porcine leydig cells cultured in vitro were collected to determine the median lethal dose (LD50) of ZEN with tetrazolium-based colorimetric assay (MTT assay). Comet assay was carried out to detect the DNA damage of porcine leydig cells exposed to at 0 (negative group), 1, 5, 10, 20, 40 μmol/L of ZEN. [Result] The percentage of cell tail was 16.67%, 34.00%, 40.67%, 52.00% and 64.67% under 0, 1, 5, 10 and 20 μmol/L of ZEN, respectively; the differences between the percentages of cell tail in various experimental groups had extremely significant statistical significance compared with the negative group (P<0.01), showing a significant dose-effect relationship; Tail length in various groups was 57.60±4.78, 57.75±6.25, 78.97±5.83, 100.50±6.94 and 146.83±12.31 μm, respectively; Tail DNA % in various groups was 21.29±2.25%, 22.24±2.43%, 31.21±6.27%, 37.45±4.33% and 60.68±9.83%, respectively; Tail length and Tail DNA % in experimental groups with ZEN concentration above 5 μmol/L showed significant differences (P<0.05) compared with the negative group, which showed an upward trend with the increase of ZEN concentration. [Conclusion] ZEN has genotoxic effect on porcine leydig cells, which can cause DNA damage, with a significant dose-effect relationship.
文摘Objective To assess the effect of benzene on sperm DNA damage ;Methods Twenty-seven benzene-exposed workers were selected as exposed group and 35 normal sperm donors as control group. Air concentration of benzene series in workshop was determined by gas chromatography. As an internal exposure dose of benzene, the concentration of trans, trans-muconic acid （ttMA） was determined by high performance liquid chromatography. DNA was detected by modified single cell gel electrophoresis （SCGE）. Results The air concentrations of benzene, toluene and xylene at the workplace were 86.49±2.83 mg/m^3, 97.20±3.52 mg/m^3 and 97.45± 2.10 mg/m^3, respectively. Urinary ttMA in exposed group （1.040 ± 0.617 mg/L） was significantly higher than that of control group （0.819 ± 0.157 mg/L）. The percentage of head DNA, determined by modified SCGE method, significantly decreased in the exposed group （n=13, 70.18% ± 7.36%） compared with the control （n=16, 90.62% ± 2.94%）（P〈0.001）. Conclusion The modified SCGE method can be used to investigate the damage of sperm DNA. As genotoxin and reprotoxins, benzene had direct effect on the germ cells during the spermatogenesiss.
基金Supported by Natural Science Foundation of Hebei Province(C2008000591)~~
文摘[Objective] The like-rocket immunoelectrophoresis was used to explore a new feasible electrophoresis method for single cell gel electrophoresis assay （comet assay）.[Method] The like-rocket immunoelectrophoresis was used for single cell gel electrophoresis assay to detect DNA damage at single cell level,then it was compared with traditional electrophoresis method to analyze its advantage and disadvantages.[Result] Under cell DNA undamaged state,the results of two electrophoresis methods were consistent.When cell DNA was damaged,the comet tail divergence of some cells under traditional electrophoresis method were drifted,however,the comet tail image of like-rocket immunoelectrophoresis was concentrated and not shifted.[Conclusion] The like-rocket immunoelectrophoresis had some advantages.
文摘DNA damage is one of the most important consequences of oxidative stress in the cells. If DNA repair is unable to modify these inducible DNA damages, genomic instability may lead to mutation, cancer, aging and many other diseases. Single cell gel electrophoresis or comet assay is a common and versatile method to quantify these types of DNA damages. DNA damages induced by hydrogen peroxide(H_2O_2) are one of the proper models for measurement of protective ability of different compounds. So the main aim of this review is to provide an overview about protection ability of medicinal plants and their potential mechanism against H_2O_2 induced DNA damages. In this review, relevant researches on the effect of medicinal plants on DNA damages induced by H_2O_2 and possible molecular mechanisms are discussed.It seems that, medicinal plants are considered as therapeutic key factors to protect DNA from consequences caused by oxidative stress. Sufficient in vitro evidences introduce them as DNA protective agents through different mechanisms including antioxidant activity and some other cellular mechanisms. Moreover, in order to correlate the antigenotoxicity effects with their potential antioxidant property, most of medicinal plants were evaluated in term of antioxidant activity using standard methods. This review highlights the preventive effects of herbal medicine against oxidative DNA damages as well as provides rational possibility to engage them in animal studies and future clinical investigations.
基金This work was supported by the Key Project of Chinese Ministry of Education(02080)the Open Fund of State Key Laboratory of Freshwater Ecology and Biotechnology,Institute of Hydrobiology,The Chinese Academy of Science(2002FB08).
文摘The effects of heavy metals Cd^(2+),Pb^(2+)and Zn^(2+)at 0.05,0.5 and 5.0 mg/L level and their interactions at 0.5 mg/L level on DNA damage in hepatopancreas of loach Misgurnus anguillicaudatus for 1-35 days exposure were examined by single cell gel electrophoresis(SCGE).For each test group,20 loaches with similar body size(5.17-7.99 g;11.79-13.21 cm)were selected and kept in aquaria with dechlori-nated water at(22±1)℃and fed a commercial diet every 48 h.According to the percentage of damaged DNA with tail and its TL/D(tail length to diameter of nucleus)value,the relationship between DNA damage degree and heavy metal dose and exposure time was determined.Results showed that the percentage of damaged DNA and the TL/D value were increased with the prolonged exposure time.The highest percentage(84.85%)of damaged DNA was shown in 5.0 mg/L Zn^(2+)group after 28 days exposure and the biggest TL/D value(2.50)in all treated groups after 35 days exposure.During the first treated week,the damnification of DNA was mainly recognized as the first level,after that time,the third damaged level was mostly observed and the percentage of damaged DNA was beyond 80%.The joint toxic effects among Cd^(2+),Pb^(2+)or Zn^(2+)revealed much complexity,but it generally displayed that the presence of Cd^(2+)could enhance the genotoxicity of Pb^(2+)or Zn^(2+).In conclusion,the results suggestedthattherewasasignificanttime-anddose-depended relationship between the heavy metal and DNA damage in hepatopancreas of loach,and SCGE could represent a useful means to evaluate the genotoxicity of environmental contamination on aquatic organisms.
