应用哺乳动物抗体检测阿维菌素抗性果蝇P糖蛋白的表达

【目的】随着杀虫剂阿维菌素的广泛应用,靶标生物对其抗性问题日益严重。以往的研究显示,细胞膜转运蛋白P糖蛋白可能与抗药性有关。但在昆虫对阿维菌素的抗性研究中,由于目前市场上缺少专门针对昆虫的商业化抗体,使得这一研究受限。为此,本研究尝试应用其他物种的抗体开展P糖蛋白的检测。【方法】以果蝇为测试昆虫,以阿维菌素作为测试药物,采用免疫印迹方法用鼠抗人P糖蛋白单克隆抗体检测阿维菌素敏感品系与阿维菌素抗性品系果蝇中的P糖蛋白的表达水平。【结果】检测出果蝇体内P糖蛋白的特异性表达,且无明显非特异性条带;与敏感品系果蝇相比,阿维菌素抗性品系果蝇P糖蛋白的表达水平明显升高。【结论】用针对人及其他脊椎动物的P糖蛋白单克隆抗体检测果蝇P糖蛋白的表达可行,且果蝇对阿维菌素的抗性可能与P糖蛋白的表达升高有关。

Expression level of P-glycoprotein in avermectin-resistant Drosophila melanogaster by using the anti-human antibody

Aim] With the widespread application of avermectin, the resistance of target organisms is becoming a more and more serious problem. Previous studies suggest that the membrane transporter P-glycoprotein (P-gp) overexpression is essential for resistance of cancer cells to chemotherapeutic drugs. However, insecticide resistance study was limited, partly because suitable commercial insect antibodies were not available.Method] In this study, we used anti-human antibody for the detection of P-gp in insects. The expression of P-gp was detected by immunoblotting in the avermectin-sensitive and avemectin-resistant strains of Drosophila melanogaster, using mouse anti-human P-gp monoclonal antibody (C219) as the primary antibody.Result] The expression of P-gp in the avermectin-resistant strain of Drosophila was higher than that in the avermectin-sensitive strain with no obvious nonspecfic bands detected.Conclusion] The expression level of P-gp in Drosophila may be related to the avermectin resistance. In addition, it is feasible to use anti-human antibody to detect the P-gp level in insects.