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《Science of Sericulture》 2011-03
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Effects of Heavy Metal Lead on Activity of Glutathione S-transferase and Glutathione Peroxidase and on Expression of Related Genes in Tissues of Bombyx mori

LI Yan-Mei1 YUAN Hong-Xia1 XU Xu1 SHU Mei-Ying1 ZHUO Wei-Wei1 SIMA Yang-Hu1,2 XU Shi-Qing1,2*(1Department of Applied Biology,Medical College of Soochow University,Suzhou Jiangsu 215123,China;2National Engineering Laboratory for Modern Silk,Suzhou Jiangsu 215123,China)  
Glutathione S-transferase(GST) and glutathione peroxidase(GSH-Px) are important detoxifying enzymes in vivo.In order to study the influencing mechanism of heavy metal lead on lepidopterous insect,we fed the Bombyx mori with artificial diet added with Pb(NO3)2 for Pb2+ exposure.Enzyme activity assay and real-time PCR were employed to investigate the activity of enzymes and the mRNA transcription level of related genes in tissues of day 6 larvae of the 5th instar.In normal day 6 larvae of the 5th instar,GST enzyme activity in gonad was 20.0 and 34.6 times of that in fat body and midgut respectively,and GSH-Px enzyme activity was 57.7 and 46.6 times of that in fat body and midgut respectively.After the larvae fed on artificial diet containing 10 mg/kg or higher Pb2+,activity of the two enzymes in gonad decreased by many times.When the artificial diet contained 20 ~80 mg/kg Pb2+,mRNA transcriptional levels of Gstd1 gene and Gsh-Px gene were also significantly down-regulated,with more significant down-regulation in males over females.After the larvae fed on artificial diet containing 10 ~160 mg/kg Pb2+,the activity of GST and GSH-Px in fat body increased significantly.Effective Pb2+ concentrations to these two enzymes could be divided into two ranges: 10 ~160 mg/kg and 20 ~80 mg/kg,respectively.However,within the range of 20 ~80 mg/kg Pb2+,mRNA transcriptions of Gsh-Px and Gstd1 were inhibited.Moreover,GSH-Px activity and mRNA level of Gsh-Px in fat body of female larvae,and GST activity and mRNA level of Gstd1 in fat body of male larvae were more sensitive to Pb2+ exposure.After 20 mg/kg or more Pb2+ exposure,GST activity in the midgut increased dramatically,but GSH-Px activity only changed slightly and there was no significant difference between male and female larvae.mRNA transcriptional level of Gstd1 gene had a trend of up-regulation,while that of Gsh-Px was significantly down-regulated.The results suggested that,although silkworm larvae have higher enzyme activity of GST and GSH-Px in gonads than in midgut and fat body,they are sensitive to toxicity of Pb2+.Nevertheless,male larvae are more sensitive than female ones,and the antioxidant defense system in gonads is quite weak in resisting toxicity from Pb2+.
【Fund】: 苏州大学重大应用研究培育项目(No.Q3034850)
【CateGory Index】: S884.97
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