title: The inhibition of activated hepatic stellate cells proliferation by arctigenin through G0/G1 phase cell cycle arrest: persistent p27(Kip1) induction by interfering with PI3K/Akt/FOXO3a signaling pathway.
abstract: Proliferation of hepatic stellate cells (HSCs) is vital for the development of fibrosis during liver injury. In this study, we describe that arctigenin (ATG), a major bioactive component of Fructus Arctii, exhibited selective cytotoxic activity via inhibiting platelet-derived growth factor-BB (PDGF-BB)-activated HSCs proliferation and arrested cell cycle at G0/G1 phase, which could not be observed in normal human hepatocytes in vitro. The cyclin-dependent kinase (CDK) 4/6 activities could be strongly inhibited by ATG through down-regulation of cyclin D1 and CDK4/6 expression in early G1 phase arrest. In the ATG-treated HSCs, the expression level of p27(Kip1) and the formation of CDK2-p27(Kip1) complex were also increased. p27(Kip1) silencing significantly attenuated the effect of ATG, including cell cycle arrest and suppression of proliferation in activated HSCs. We also found that ATG suppressed PDGF-BB-induced phosphorylation of Akt and its downstream transcription factor Forkhead box O 3a (FOXO3a), decreased binding of FOXO3a to 14-3-3 protein, and stimulated nuclear translocation of FOXO3a in activated HSCs. Furthermore, knockdown of FOXO3a expression by FOXO3a siRNA attenuated ATG-induced up-regulation of p27(Kip1) in activated HSCs. All the above findings suggested that ATG could increase the levels of p27(Kip1) protein through inhibition of Akt and improvement of FOXO3a activity, in turn inhibited the CDK2 kinase activity, and eventually caused an overall inhibition of HSCs proliferation.
authors.affiliation: College of Pharmacy and Bioengineering, Chongqing University of Technology, 69 Hong Guang Avenue, Chongqing 400054, PR China. Electronic address: ao_li@cqut.edu.cn.
authors.forename: Ao
authors.initials: A
authors.lastname: Li
authors.affiliation: College of Pharmacy and Bioengineering, Chongqing University of Technology, 69 Hong Guang Avenue, Chongqing 400054, PR China.
authors.forename: Jun
authors.initials: J
authors.lastname: Wang
authors.affiliation: Institute of Life Sciences, Chongqing Medical University, 1 Yi Xue Yuan Road, Chongqing 400016, PR China.
authors.forename: Mingjun
authors.initials: M
authors.lastname: Wu
authors.affiliation: College of Pharmacy and Bioengineering, Chongqing University of Technology, 69 Hong Guang Avenue, Chongqing 400054, PR China.
authors.forename: Xiaoxun
authors.initials: X
authors.lastname: Zhang
authors.affiliation: College of Pharmacy and Bioengineering, Chongqing University of Technology, 69 Hong Guang Avenue, Chongqing 400054, PR China.
authors.forename: Hongzhi
authors.initials: H
authors.lastname: Zhang
mesh_terms: D048948:14-3-3 Proteins / Q000378:metabolism; D021581:Active Transport, Cell Nucleus / Q000187:drug effects; D000077214:Becaplermin; D002460:Cell Line; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D019938:Cyclin D1 / Q000378:metabolism; D051358:Cyclin-Dependent Kinase 4 / Q000378:metabolism; D051361:Cyclin-Dependent Kinase 6 / Q000378:metabolism; D050760:Cyclin-Dependent Kinase Inhibitor p27 / Q000378:metabolism*; D015536:Down-Regulation / Q000187:drug effects; D000071316:Forkhead Box Protein O3; D051858:Forkhead Transcription Factors / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D005663:Furans / Q000494:pharmacology*; D059585:G1 Phase Cell Cycle Checkpoints / Q000187:drug effects*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D055785:Gene Knockdown Techniques; D055166:Hepatic Stellate Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D017705:Lignans / Q000494:pharmacology*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D020574:Proto-Oncogene Proteins c-sis / Q000494:pharmacology; D034741:RNA, Small Interfering / Q000235:genetics; D016192:Resting Phase, Cell Cycle / Q000187:drug effects*; D015398:Signal Transduction / Q000187:drug effects*
chemical_list: D048948:14-3-3 Proteins; C499517:FOXO3 protein, human; D000071316:Forkhead Box Protein O3; D051858:Forkhead Transcription Factors; D005663:Furans; D017705:Lignans; D020574:Proto-Oncogene Proteins c-sis; D034741:RNA, Small Interfering; D019938:Cyclin D1; D050760:Cyclin-Dependent Kinase Inhibitor p27; D000077214:Becaplermin; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt; D051358:Cyclin-Dependent Kinase 4; D051361:Cyclin-Dependent Kinase 6; C071942:arctigenin
keywords: 5-Bromo-2-deoxyuridine (PubChem CID: 6035); Arctigenin; Arctigenin (PubChem CID: 64981); Dimethyl sulfoxide (PubChem CID: 679); Forkhead box O (FOXO); Hepatic stellate cell; LY294002 (PubChem CID: 3973); Liver fibrosis; PHT-427 (PubChem CID: 44240850); Phenylmethanesulfonyl fluoride (PubChem CID: 4784); Proliferation; Propidium iodide (PubChem CID: 104981); SB203580 (PubChem CID: 176155); Sodium dodecyl sulfate (PubChem CID: 3423265); U0126 (PubChem CID: 3006531); p27(Kip1)