Besides structural modification of natural bioactive products to afford promising agrochemical candidates, investigation of their mechanisms of action against pests is also an important strategy to obtain novel potentially botanical pesticides. N-(p-Ethyl)phenylsulfonylmatrinic acid (2), derived from an natural alkaloid matrine (1), exhibited about 5.9-fold more pronounced acaricidal activity than 1 against the adult females of Tetranychus cinnabarinus Boisduval, and good control efficiency in the greenhouse. By comparison of nAChR, AChE and VGSC of treated and untreated T. cinnabarinus via RT-PCR and qRT-PCR analysis, it was found that compound 2 could activate nAChR and VGSC via up-regulation of nAChR alpha1, alpha4 and alpha5 subunits and VGSC expressions; compound 2 may be the AChE and AChE enzyme inhibitor. Importantly, a scheme of compound 2 interaction with nAChR, AChE and VGSC of T. cinnabarinus was proposed. It will lay the foundation for future optimization and application of matrine derivatives as agrochemicals. CI - Copyright (c) 2020 Elsevier Ltd. All rights reserved.