Department of Human Biology, Punjabi University, Punjab, India.
Rejuvenation Res. 2010 Apr-Jun;13(2-3):281-4. doi: 10.1089/rej.2009.0931.
The main aim of this study was to evaluate genotoxic effects of pesticides in association with glutathione S-transferase (GST) polymorphism. To achieve this aim, DNA damage and the genotypes of the GSTM1 and GSTT1 genes were studied from blood lymphocytes of pesticide-exposed and unexposed (control) agricultural workers of the Punjab region of northwestern India. The blood samples were collected from 40 exposed and 27 unexposed subjects from the Kakrala and Sanour villages of Patiala district. DNA damage was evaluated by using an alkaline comet assay. The analysis of the comets was done through visual scoring and image analysis software (Tritek's CometScore). Damage Index (DI), Damage Frequency (DF) (calculated by visual scoring method), and % DNA in tail (measured by image analysis software) were considered for assessing DNA damage. The DNA extraction from blood cells was done using proteinase K and the phenol-chloroform method, and genotyping of GSTM1 and GSTT1 was done using multiplex PCR. It was found that all the pesticide-exposed subjects showed higher DI, DF, and % DNA in tail in comparison to the controls. The statistical comparison of DNA damage between the exposed group and unexposed group revealed highly significant differences (p < 0.05; Mann-Whitney U-test). In addition, the GSTT1 gene deletion and simultaneous deletions of GSTM1 and GSTT1 genes in increasing DNA damage were observed in the exposed group.
本研究的主要目的是评估农药与谷胱甘肽 S-转移酶 (GST) 多态性联合的遗传毒性作用。为了实现这一目标，研究了来自印度西北部旁遮普地区接触和未接触（对照）农药的农业工人的血淋巴细胞中的 DNA 损伤和 GSTM1 和 GSTT1 基因的基因型。从 Patiala 区 Kakrala 和 Sanour 村的 40 名暴露组和 27 名未暴露组采集血样。使用碱性彗星试验评估 DNA 损伤。通过肉眼评分和图像分析软件（Tritek 的 CometScore）对彗星进行分析。采用视觉评分法计算损伤指数（DI）、损伤频率（DF）（通过肉眼评分法计算）和尾部的 %DNA（通过图像分析软件测量），以评估 DNA 损伤。使用蛋白酶 K 和酚-氯仿法从血细胞中提取 DNA，并使用多重 PCR 进行 GSTM1 和 GSTT1 基因分型。结果发现，与对照组相比，所有接触农药的受试者的 DI、DF 和尾部的 %DNA 均升高。暴露组和未暴露组之间的 DNA 损伤的统计学比较显示差异具有高度显著性（p < 0.05；Mann-Whitney U 检验）。此外，在暴露组中观察到 GSTT1 基因缺失以及 GSTM1 和 GSTT1 基因同时缺失与 DNA 损伤增加有关。
