title: An Ingenol Derived from Euphorbia kansui Induces Hepatocyte Cytotoxicity by Triggering G0/G1 Cell Cycle Arrest and Regulating the Mitochondrial Apoptosis Pathway in Vitro.
abstract: Natural product lingenol, a purified diterpenoid compound derived from the root of Euphorbia kansui, exerts serious hepatotoxicity; however, the molecular mechanisms remain to be defined. In the present study, cell counting Kit-8 (CCK-8), inverted phase contrast microscope and flow cytometry were used to demonstrate that lingenol significantly inhibited L-O2 cells proliferation, and induced cell cycle arrest and apoptosis. Moreover, the results investigated that lingenol markedly disrupted mitochondrial functions by high content screening (HCS). In addition, the up-regulation of cytochrome c, AIF and Apaf-1 and activation of caspases were found in L-O2 cells detected by Western blotting and ELISA assay, which was required for lingenol activation of cytochrome c-mediated caspase cascades and AIF-mediated DNA damage. Mechanistic investigations revealed that lingenol significantly down-regulated the Bcl-2/Bax ratio and enhanced the reactive oxygen species (ROS) in L-O2 cells. These data collectively indicated that lingenol modulation of ROS and Bcl-2/Bax ratio led to cell cycle arrest and mitochondrial-mediated apoptosis in L-O2 cells in vitro. All of these results will be helpful to reveal the hepatotoxicity mechanism of Euphorbia kansui and to effectively guide safer and better clinical application of this herb.
authors.affiliation: Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, Jiangsu Key Laboratory for High Technology Research of TCM Formulae, Nanjing University of Chinese Medicine, Nanjing 210023, China. yanxiaojing963@163.com.|Changzhou Affiliated Hospital of Nanjing University of Chinese Medicine, Changzhou 213003, China. yanxiaojing963@163.com.
authors.forename: Xiaojing
authors.initials: X
authors.lastname: Yan
authors.affiliation: Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, Jiangsu Key Laboratory for High Technology Research of TCM Formulae, Nanjing University of Chinese Medicine, Nanjing 210023, China. zhangli@njutcm.edu.cn.
authors.forename: Li
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, Jiangsu Key Laboratory for High Technology Research of TCM Formulae, Nanjing University of Chinese Medicine, Nanjing 210023, China. raindc@163.com.
authors.forename: Yudan
authors.initials: Y
authors.lastname: Cao
authors.affiliation: Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, Jiangsu Key Laboratory for High Technology Research of TCM Formulae, Nanjing University of Chinese Medicine, Nanjing 210023, China. njweifengyao@163.com.
authors.forename: Weifeng
authors.initials: W
authors.lastname: Yao
authors.affiliation: Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, Jiangsu Key Laboratory for High Technology Research of TCM Formulae, Nanjing University of Chinese Medicine, Nanjing 210023, China. yupingtang@njutcm.edu.cn.
authors.forename: Yuping
authors.initials: Y
authors.lastname: Tang
authors.affiliation: Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, Jiangsu Key Laboratory for High Technology Research of TCM Formulae, Nanjing University of Chinese Medicine, Nanjing 210023, China. awding105@163.com.
authors.forename: Anwei
authors.initials: A
authors.lastname: Ding
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D051033:Apoptosis Inducing Factor / Q000096:biosynthesis; D053477:Apoptotic Protease-Activating Factor 1 / Q000096:biosynthesis; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects*; D045304:Cytochromes c / Q000096:biosynthesis; D004224:Diterpenes / Q000008:administration & dosage / Q000009:adverse effects*; D028481:Euphorbia / Q000737:chemistry; D005786:Gene Expression Regulation / Q000187:drug effects; D022781:Hepatocytes / Q000187:drug effects; D008928:Mitochondria / Q000187:drug effects; D010936:Plant Extracts / Q000008:administration & dosage* / Q000009:adverse effects / Q000737:chemistry; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: C494829:AIFM1 protein, human; C506592:APAF1 protein, human; D051033:Apoptosis Inducing Factor; D053477:Apoptotic Protease-Activating Factor 1; D004224:Diterpenes; D010936:Plant Extracts; D017382:Reactive Oxygen Species; D045304:Cytochromes c; C008125:ingenol
keywords: Bcl-2/Bax; ROS; cell apoptosis; cell cycle arrest; hepatotoxicity; high content screen; lingenol