›› 2011, Vol. 54 ›› Issue (2): 117-126.doi:

• 研究论文 •    下一篇

亚洲玉米螟幼虫应对大肠杆菌注射的血淋巴免疫应激反应

冯从经, 董秋安, 翟会峰, 陈根宝, 杨俊明, 苗俊玲   

  • 出版日期:2011-02-20 发布日期:2011-03-10
  • 通讯作者: 冯从经

Immunological and stress response of the hemolymph of Ostrinia furnacalis Guenée (Lepidoptera: Pyralidae) larvae to the injection of Escherichia coli

FENG Cong-Jing, DONG Qiu-An, ZHAI Hui-Feng, CHEN Gen-Bao, YANG Jun-Ming, MIAO Jun-Ling   

  • Online:2011-02-20 Published:2011-03-10

摘要: 为研究大肠杆菌Escherichia coli侵染引发亚洲玉米螟Ostrinia furnacalis Guenée幼虫免疫应激反应的机理,本实验测定了分别注射生理盐水以及3×1033×104,3×1053×106个细胞/mL大肠杆菌后亚洲玉米螟5龄幼虫血淋巴中血细胞总数(THC)、颗粒细胞和浆血细胞数量,血清中酚氧化酶(PO)、谷胱甘肽过氧化物酶(GSH-px)、谷胱甘肽还原酶(GR)和谷胱甘肽-S-转移酶(GST)的活性,通过流式细胞仪分析了血细胞活性氧自由基(ROS)水平的动态变化。结果表明:与对照组相比,注射3×105和3×106个细胞/mL大肠杆菌细胞后12 h,可引起亚洲玉米螟5龄幼虫THC及浆血细胞、颗粒细胞数量明显上升(P<0.01),同时应激产生大量ROS。3×104,3×105和3×106个细胞/mL大肠杆菌3个不同浓度处理组均引起幼虫体内PO活性显著升高(P<0.01),诱导幼虫血清中GSH-px,GST及GR的活性上升(P<0.01)。这些结果表明, 亚洲玉米螟幼虫受到大肠杆菌侵染后,其血淋巴细胞免疫和体液免疫能力受到显著影响, 可诱导血清中GSH-px,GST和GR活性升高以清除过多的ROS, 防止其毒害。

关键词: 亚洲玉米螟, 大肠杆菌, 血细胞, 酚氧化酶, 抗氧化酶, 活性氧自由基

Abstract:  In order to explore the mechanism of immunological and stress response of Ostrinia furnacalis Guenée larvae invaded by Escherichia coli, the numbers of total hemocytes, granular hemocytes and plasmatocytes in the 5th instar larvae of O. furnacalis injected with physiological saline, 3×103, 3×104, 3×105 and 3×106 cells/mL E. coli, respectively, were counted, and the activities of phenoloxidase (PO), glutathione peroxidase (GSH-px), glutathione-S-transferase (GST) and glutathione reductase (GR) were determined. The content of reactive oxygen species (ROS) of the hemocytes was also analyzed by flow cytometry (FACS). The results showed that the numbers of total hemocytes, granular hemocytes and plasmatocytes in the 5th instar larvae of O. furnacalis increased significantly in 12 h post injection with 3×105 cells/mL and 3×106 cells/mL E. coli (P<0.01), compared with the control group. The content of ROS in hemocytes also increased at the same time. Injections with 3×104, 3×105 and 3×106 cells/mL E. coli triggered the enhancement of PO activity, and also induced the increase of the activities of GSH-px, GST, and GR in the serum. The results suggest that the abilities of cellular and humoral immunity of O. furnacalis larvae are significantly affected by the injection of E. coli, and the activities of GSH-px, GST and GR are also induced to scavenge the excessive ROS to prevent from the toxic effect of ROS.

Key words: Ostrinia furnacalisEscherichia coli;hemocyte, phenoloxidase, antioxidant enzymes, reactive oxygen species