title: A High Content Screening Assay to Identify Compounds with Anti-Epithelial-Mesenchymal Transition Effects from the Chinese Herbal Medicine Tong-Mai-Yang-Xin-Wan.
abstract: Chronic kidney disease (CKD) is a worldwide health problem with growing prevalence in developing countries. Renal tubular epithelial-mesenchymal transition (EMT) is a critical step and key factor in the development of this condition. Renal tubulointerstitial fibrosis is a basic pathological change at the later stages of the disease. Therefore, blocking the development of EMT could be a critical factor in curing CKD. We have established a cell-based high-content screening (HCS) method to identify inhibitors of EMT in human proximal tubular epithelial (HK-2) cells by automatic acquisition and processing of dual-fluorescent labeled images. With the aid of chromatographic separation and mass spectrometry, we achieved the rapid and reliable screening of active compounds from the Chinese herbal medicine Tong-Mai-Yang-Xin-Wan (TMYX) for treating EMT. Five fractions were found to exert anti-EMT activity and were further identified by liquid chromatography coupled with tandem mass spectrometry. Glycyrrhizic acid, glyasperin A, and licorisoflavan A were found to inhibit EMT. The proposed approach was successfully applied to screen active compounds from TMYX on TGF-β1-stimulated HK-2 cells and may offer a new means for identifying lead compounds for treating EMT from registered Chinese herbal medicines.
authors.affiliation: TCM Research Center, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China. 18867141533@163.com.
authors.forename: Ningning
authors.initials: N
authors.lastname: Liu
authors.affiliation: TCM Research Center, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China. mooching@126.com.
authors.forename: Lailai
authors.initials: L
authors.lastname: Li
authors.affiliation: Zhejiang Cancer Research Institute, Zhejiang Cancer Hospital, Hangzhou 310022, China. zhuxin@zjcc.org.cn.
authors.forename: Xin
authors.initials: X
authors.lastname: Zhu
authors.affiliation: Zhejiang Cancer Research Institute, Zhejiang Cancer Hospital, Hangzhou 310022, China. lingzq@hotmail.com.
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Ling
authors.affiliation: Zhejiang Cancer Research Institute, Zhejiang Cancer Hospital, Hangzhou 310022, China. fengjg@zjcc.org.cn.
authors.forename: Jianguo
authors.initials: J
authors.lastname: Feng
authors.affiliation: Department of Nephrology, The Second Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310029, China. huyinghz@126.com.
authors.forename: Ying
authors.initials: Y
authors.lastname: Hu
authors.affiliation: TCM Research Center, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China. vip_wangyi@126.com.
authors.forename: Yi
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Nephrology, The Second Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310029, China. tiantangmou@126.com.
authors.forename: Lijun
authors.initials: L
authors.lastname: Mou
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China. mysky@zju.edu.cn.
authors.forename: Yi
authors.initials: Y
authors.lastname: Wang
mesh_terms: D015415:Biomarkers; D002460:Cell Line; D002845:Chromatography / Q000379:methods; D055808:Drug Discovery; D004365:Drugs, Chinese Herbal / Q000737:chemistry / Q000494:pharmacology*; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects*; D015870:Gene Expression; D057166:High-Throughput Screening Assays; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods; D015203:Reproducibility of Results
chemical_list: D015415:Biomarkers; D004365:Drugs, Chinese Herbal
keywords: epithelial mesenchymal transition; fluorescence imaging; high-content screening