The aryl hydrocarbon receptor(AhR)plays a crucial role in regulating many physiological processes.Activating the AhReCYP1A1 axis has emerged as a novel therapeutic strategy against various inflammatory diseases.Here,a...The aryl hydrocarbon receptor(AhR)plays a crucial role in regulating many physiological processes.Activating the AhReCYP1A1 axis has emerged as a novel therapeutic strategy against various inflammatory diseases.Here,a practical in situ cell-based fluorometric assay was constructed to screen AhR-CYP1A1 axis modulators,via functional sensing of CYP1A1 activities in live cells.Firstly,a cell-permeable,isoform-specific enzyme-activable fluorogenic substrate for CYP1A1 was rationally constructed for in-situ visualizing the dynamic changes of CYP1A1 function in living systems,which was subsequently used for discovering the efficacious modulators of the AhReCYP1A1 axis.Following screening of a compound library,LAC-7 was identified as an efficacious activator of the AhReCYP1A1 axis,which dose-dependently up-regulated the expression levels of both CYP1A1 and AhR in multiple cell lines.LAC-7 also suppressed macrophage M1 polarization and reduced the levels of inflammatory factors in LPS-induced bone marrow-derived macrophages.Animal tests showed that LAC-7 could significantly mitigate DSS-induced ulcerative colitis and LPS-induced acute lung injury in mice,and markedly reduced the levels of multiple inflammatory factors.Collectively,an optimized fluorometric cell-based assay was devised for in situ functional imaging of CYP1A1 activities in living systems,which strongly facilitated the discovery of efficacious modulators of the AhReCYP1A1 axis as novel antiinflammatory agents.展开更多
基金supported by National Natural Science Foundation of China(Nos.U23A20516,81922070,82273897 and 52303191)National Key Research and Development Program of China(No.2022YFC3502000)+5 种基金Shanghai Science and Technology Innovation Action Plans(21S21900600,China)the Organizational Key Research and Development Program of Shanghai University of Traditional Chinese Medicine(No.2023YZZ02,China)Shanghai Municipal Health Commission’s TCM research project(No.2022CX005,China)Innovation Team and Talents Cultivation Program of National Administration of Traditional Chinese Medicine(No.ZYYCXTDD-202004,China)the State Key Laboratory of Fine Chemicals,Dalian University of Technology(No.KF 2202,China)Shanghai University of Traditional Chinese Medicine(No.2021LK023,China).
文摘The aryl hydrocarbon receptor(AhR)plays a crucial role in regulating many physiological processes.Activating the AhReCYP1A1 axis has emerged as a novel therapeutic strategy against various inflammatory diseases.Here,a practical in situ cell-based fluorometric assay was constructed to screen AhR-CYP1A1 axis modulators,via functional sensing of CYP1A1 activities in live cells.Firstly,a cell-permeable,isoform-specific enzyme-activable fluorogenic substrate for CYP1A1 was rationally constructed for in-situ visualizing the dynamic changes of CYP1A1 function in living systems,which was subsequently used for discovering the efficacious modulators of the AhReCYP1A1 axis.Following screening of a compound library,LAC-7 was identified as an efficacious activator of the AhReCYP1A1 axis,which dose-dependently up-regulated the expression levels of both CYP1A1 and AhR in multiple cell lines.LAC-7 also suppressed macrophage M1 polarization and reduced the levels of inflammatory factors in LPS-induced bone marrow-derived macrophages.Animal tests showed that LAC-7 could significantly mitigate DSS-induced ulcerative colitis and LPS-induced acute lung injury in mice,and markedly reduced the levels of multiple inflammatory factors.Collectively,an optimized fluorometric cell-based assay was devised for in situ functional imaging of CYP1A1 activities in living systems,which strongly facilitated the discovery of efficacious modulators of the AhReCYP1A1 axis as novel antiinflammatory agents.
