›› 2011, Vol. 54 ›› Issue (1): 20-26.doi:

• 研究论文 • 上一篇    下一篇

芸香苷对家蚕谷胱甘肽-S-转移酶部分基因的诱导表达

张婷, 卫正国, 高瑞娜, 王瑞娴, 赵国栋, 李兵, 沈卫德   

  • 出版日期:2011-01-20 发布日期:2011-01-20
  • 通讯作者: 沈卫德

Induction of expression of partial glutathione-S-transferase genes in Bombyx mori by rutin

ZHANG Ting, WEI Zheng-Guo, GAO Rui-Na, WANG Rui-Xian, ZHAO Guo-Dong, LI Bing, SHEN Wei-De   

  • Online:2011-01-20 Published:2011-01-20

摘要: 谷胱甘肽-S-转移酶(glutathione-S-transferases, GSTs) 是对一种机体的解毒代谢起重要作用并可以被诱导的酶系。为了从分子水平上探究家蚕Bombyx mori GSTs 基因与植物次生物质芸香苷代谢的关系, 本实验采用双跟踪标定定量PCR (dual-spike-in qPCR)方法, 用 5×10-1, 5×10-2, 5×10-3 ng/μL 3 个浓度芸香苷溶液处理家蚕5龄幼虫, 并对各组织中 GSTs Epsilon 家族不同基因的转录水平进行检测。结果显示, 浓度为 5×10-2 ng/μL 的芸香苷溶液能诱导GSTs 基因的转录水平发生显著变化。在中肠中, BmGSTe1, BmGSTe2 和 BmGSTe6的诱导转录水平较高, 在诱导后24 h达到最大值; 在脂肪体中BmGSTe1, BmGSTe6和BmGSTe7的转录水平相对较高且分别在诱导后 2 h, 2 h和4 h 达到最大值; 而GSTs基因在马氏管中表达量均很低或者检测不到表达。与 5×10-2 ng/μL 浓度相比, 5×10-1 ng/μL 的芸香苷溶液诱导后各基因的转录水平上升的幅度较小并且达到峰值的时间不同, 而 5×10-3 ng/μL 浓度的芸香苷溶液并不能使GSTs基因的诱导转录水平发生变化。结果提示, 家蚕 GSTs Epsilon 家族基因对芸香苷的代谢具有重要作用。

关键词: 家蚕, 谷胱甘肽-S-转移酶基因, 芸香苷, 双跟踪标定定量PCR, 诱导表达

Abstract: Glutathione-S-transferases (GSTs) play an important role in detoxification and can be induced. In order to study the relationship between the metabolism of the plant secondary metabolite rutin and the GSTs genes of Bombyx mori, we detected the transcription levels of Epsilon-class GSTs genes in different tissues of the 5th instar larvae and those treated with rutin (5×10-1, 5×10-2 and 5×10-3 ng/μL) using dual-spike-in qPCR. The results showed that rutin (5×10-2 ng/μL) can induce a significant change of the transcription levels of GSTs genes. In the midgut, BmGSTe1, BmGSTe2 and BmGSTe6 had higher transcription levels and reached the maximum at 24 h after induction; in fat body, however, BmGSTe1, BmGSTe6 and BmGSTe7 had higher transcription levels and reached the peak at 2 h, 2 h and 4 h after induction, respectively; and the transcription level of the GSTs genes did not change significantly or was undetectable in Malpighian tubules. Compared with the treatment with 5×10-2 ng/μL rutin, 5×10-1 ng/μL rutin could induce a little change of the transcription levels of GSTs genes and the time when the expression peak occurred was not the same. However, 5×10-3 ng/μL rutin could not induce the change of the transcription level of the GSTs genes. The results suggest that the Epsilon-class GSTs genes play an important role in the metabolism of rutin.

Key words: Bombyx mori, GSTs genes, rutin, dual spike-in qPCR, induced expression