淡色库蚊表皮蛋白对残杀威抗性的蛋白组学研究

doi:10.20199/j.issn.1672-2302.2025.03.009

摘要/Abstract

摘要：

目的探究淡色库蚊对残杀威抗性的表皮蛋白适应机制，明确差异表达蛋白在抗性与敏感株系抗性形成中的作用。方法以实验室淡色库蚊敏感株系为亲本，连续20代残杀威筛选培育抗性株系，收集抗性与敏感株淡色库蚊各发育阶段样本。采用同位素标记相对和绝对定量技术（isobaric tags for relative and absolute quantification, iTRAQ）定量分析蛋白组，利用COG、GO和KEGG数据库进行功能注释及通路富集分析，并通过平行反应监测验证关键蛋白表达。结果共鉴定出156个差异表达蛋白（59个上调，97个下调），淡色库蚊抗性株系中表皮结构成分蛋白（B0WIY4、B0W0L1）、几丁质结合蛋白及幼虫表皮蛋白显著上调；细胞骨架相关蛋白（B0WY76）、能量代谢蛋白（丙酮酸脱氢酶B0XA87）及核糖体蛋白（B0WQU6）显著下调。平行反应监测验证了肌动蛋白（B0WY76）等11类蛋白表达。结论淡色库蚊对残杀威抗性与表皮蛋白差异表达和细胞骨架重构引起的表皮结构改变密切相关，这种结构改变可能延缓杀虫剂表皮渗透，从而增强对残杀威的抗性。

关键词: 淡色库蚊, 残杀威, 抗性机制, 表皮蛋白, 蛋白质组学

Abstract:

Objective To investigate the potential cuticular protein adaptation mechanisms of Culex pipiens pallens resistance to propoxur for characterizing the functional roles of differentially expressed proteins (DEPs) in conferring the resistance between resistant and susceptible strains. Methods A propoxur-resistant strain was generated through 20 generations of selective pressure from a laboratory-maintained susceptible strain. Samples spanning multiple developmental stages from both strains were subjected to quantitative proteomic profiling using isobaric tags for relative and absolute quantification (iTRAQ). Subsequent functional annotation and pathway enrichment analyses were performed by means of COG, GO, and KEGG databases, followed by targeted validation of pivotal proteins via parallel reaction monitoring (PRM). Results A total of 156 DEPs, comprising 59 upregulated and 97 downregulated proteins, were identified. Cuticular structural components (B0WIY4, B0W0L1), chitin-binding proteins, and larval cuticular proteins exhibited marked upregulation in the resistant strain, whereas cytoskeleton-associated proteins (B0WY76), energy metabolism regulators (pyruvate dehydrogenase B0XA87), and ribosomal proteins (B0WQU6) were significantly downregulated. PRM analysis confirmed the expression trends of 11 representative proteins, including actin (B0WY76). Conclusion The resistance of Culex pipiens pallens to propoxur is closely related to the structural alterations of the cuticle caused by differential expression of cuticular proteins and cytoskeletal remodeling. These alterations likely impede insecticide penetration through the cuticle, thereby enhancing propoxur resistance.

Key words: Culex pipiens pallens, Propoxur, Resistant mechanism, Cuticular protein, Proteomics

中图分类号:

R184.31

张辉红, 徐颢源, 李文璇, 郭秀霞, 程鹏, 王海防, 刘丽娟, 张崇星. 淡色库蚊表皮蛋白对残杀威抗性的蛋白组学研究[J]. 热带病与寄生虫学, 2025, 23(3): 176-182.

ZHANG Huihong, XU Haoyuan, LI Wenxuan, GUO Xiuxia, CHENG Peng, WANG Haifang, LIU Lijuan, ZHANG Chongxing. Study on the epidermal adaptability mechanism of Culex pipiens pallens resistance to propoxur by comparative proteomics[J]. Journal of Tropical Diseases and Parasitology, 2025, 23(3): 176-182.

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GO功能分类	调节模式	GO功能类别	差异蛋白数量	代表性通路/功能	富集倍数（Fold charge, FC）
生物过程	上调	氮化合物代谢过程	12	几丁质代谢过程	5.74
		有机氮化合物代谢过程	11	糖基化合物代谢过程	3.91
		碳水化合物衍生物代谢过程	7	核苷代谢过程	3.91
	下调	细胞氮化合物生物合成过程	8	基于微管的运动	0.11
	下调	细胞氮化合物生物合成过程	8	细胞或亚细胞成分的运动	0.12
分子功能	上调	几丁质结合蛋白	3	几丁质结合	6.13
	下调	结构分子活性	18	rRNA结合	0.12
		表皮结构成分	13	脂质转运体活性	0.13
		表皮结构成分	13	表皮结构成分	0.19
细胞构成	上调	细胞外区域	4	细胞外区域	3.36
	下调	大分子复合体	12	蛋白质-DNA复合物	0.11
				核小体	0.11
				DNA包装复合体	0.11

蛋白COG类型	蛋白数据ID	蛋白结构域描述	鉴定到的肽段序列
细胞骨架	B0WY76	肌动蛋白	VAPEEHPILLTEAPLNPK
细胞骨架	B0X3L6	无脊椎动物原肌球蛋白	ALLCEQQAR
能量产生与转换	B0XA87	丙酮酸脱氢酶	VFILGEEVAQYDGAYK
	B0WXM0	泛醌醇-细胞色素c还原酶复合物核心蛋白	ADMVAAINGVSTSDVQSVAR
	B0WUY1	NADH泛醌氧化还原酶B8亚单位	EFVSSQYAALK
	B0WY92	细胞色素c氧化酶-亚单位VIb	FPNTNQTK
	B0XGQ7	线粒体NADH脱氢酶铁硫蛋白8	GAAVTLAHIFK
翻译、核糖体结构和生物发生	B0W289	40S核糖体蛋白S21	DHASIQINIVDVHPQTGR
	B0WQU6	核糖体蛋白 L37	HTCSQCGYPSAK
	B0WU22	酸性核糖体蛋白P1	AANVDIEPYWPGLFAK
翻译后修饰，蛋白质周转，伴侣蛋白	B0WTL4	蛋白酶体内肽酶复合物	EAINLALSTLK
翻译后修饰，蛋白质周转，伴侣蛋白	B0WGF9	硫氧还蛋白还原酶1	ENVEVYHAYYKPTEFFVPQR
氨基酸转运与代谢	B0WZS2	吡咯啉-5-羧酸还原酶	LAAQTVMGAGK
	B0X5I3	水解酶	LVVNETSVFTR
	B0WJC4	d-氨基酸氧化酶	LTTDPAGYPDPAWR
碳水化合物运输和代谢	B0WEB5	甘油醛-3-磷酸脱氢酶	LISWYDNEFGYSNR
碳水化合物运输和代谢	B0W5W4	丙糖磷酸异构酶	ASITELCK
信号转导机制	B0WEM1	GTP结合蛋白128up	GGGGASEQGFEVSK
	B0WCT7	磷酸酶2Cβ	NLLQSIIR
	B0W6W0	肌钙蛋白C	TGSISSETVAEILR
细胞内运输、分泌和囊泡运输	B0WUR5	分泌载体相关膜蛋白	ATVQSQFNQNR
核苷酸转运和代谢	B0X890	尿苷磷酸化酶	LPAGTQLQDISAFSYR
辅酶转运与代谢	B0W244	腺苷甲硫氨酸合酶	DTVQHIGYDDSSK
脂质运输和代谢	B0WY25	3-羟基异丁酰辅酶a水解酶	SLVEAGPVPESR