title: Interaction of AR and iNOS in lens epithelial cell: A new pathogenesis and potential therapeutic targets of diabetic cataract.
abstract: Although there is significant interest in revealing the role of aldose reductase (AR) and inducible nitric oxide synthase (iNOS) in diabetic cataract (DC), the interaction of AR and iNOS remains unknown. The aim of this study is to investigate the pathogenesis mechanisms and explore as a new potential therapeutic targets for DC. This study investigated the interaction of AR-iNOS through the methods of enzyme kinetics, molecular docking and molecular dynamics simulation, co-immunoprecipitation and fluorescence resonance energy transfer (FRET). The IC50 of AR for inhibition of iNOS activity is 0.04 μM, and the IC50 of iNOS for inhibition of AR activity is 0.042 μM through enzyme kinetics; the interface showed that ARG99 on AR and GLU317 on iNOS played the key roles in the interaction of AR-iNOS predicted by molecular docking and molecular dynamics simulation. Co-immunoprecipitation of protein complexes in human lens epithelial cell (HLEC) demonstrated that AR could association with iNOS in cell; and the interaction distance of AR-iNOS was 6.50 ± 0.22 nm detected by FRET. This study exhibited a direct inhibition interaction between AR and iNOS in HLECs. It is the first report of inhibition interaction between AR and iNOS, suggesting a new pathophysiological mechanism and providing a new insight into the therapeutic mechanism of DC.
authors.affiliation: School of Basic Medical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, 510006, China. Electronic address: xiaoxuesnowflak@163.com.
authors.forename: Xue
authors.initials: X
authors.lastname: Li
authors.affiliation: School of Bioscience and Bioengineering, South China University of Technology, Guangzhou 510006, China. Electronic address: 710304675@qq.com.
authors.forename: Wenping
authors.initials: W
authors.lastname: Liu
authors.affiliation: School of Basic Medical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, 510006, China. Electronic address: 504782301@qq.com.
authors.forename: Xinduo
authors.initials: X
authors.lastname: Huang
authors.affiliation: Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA. Electronic address: xiong@helix.mgh.harvard.edu.
authors.forename: Jianping
authors.initials: J
authors.lastname: Xiong
authors.affiliation: School of Basic Medical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, 510006, China; Antioxidants Research Laboratory, Jean Mayer USDA Human Nutrition Research Center on Aging, Tufts University, Boston, MA 02111, USA. Electronic address: jidewowxy2@163.com.
authors.forename: Xiaoyong
authors.initials: X
authors.lastname: Wei
mesh_terms: D002386:Cataract / Q000150:complications / Q000378:metabolism*; D048909:Diabetes Complications / Q000378:metabolism*; D004847:Epithelial Cells / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D020128:Inhibitory Concentration 50; D007700:Kinetics; D007908:Lens, Crystalline / Q000166:cytology; D008856:Microscopy, Fluorescence; D008968:Molecular Conformation; D062105:Molecular Docking Simulation; D056004:Molecular Dynamics Simulation; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism*; D011944:Receptors, Androgen / Q000378:metabolism*
chemical_list: C410798:AR protein, human; D011944:Receptors, Androgen; C496319:NOS2 protein, human; D052247:Nitric Oxide Synthase Type II
keywords: Aldose reductase; Diabetic cataract; Inducible nitric oxide synthase; Interaction