title: Anti-proliferative and pro-apoptotic activities of Alpinia oxyphylla on HepG2 cells through ROS-mediated signaling pathway.
abstract: BACKGROUND
  Fructus Alpiniae oxyphyllae (A. oxyphylla) is a traditional herb which is widely used in East Asian for the treatment of dyspepsia, diarrhea, abdominal pain, poor memory, inflammatory conditions and cancer.
  
  
  METHODS
  The cytotoxic activities of ethanol extract (EE) and five extract layers including petroleum ether (PE), dichloromethane (DCLM), acetoacetate (EtOAc), n-Butanol (n-Bu) and water fractions (WF) of A. oxyphylla were tested on HepG2, SW480, MCF-7, K562 and HUVEC cell lines using MTT assay and LDH release assay. The component analysis was performed on HPLC with gradient elution. Hoechst 33342 staining, DCFH-DA fluorescence microscopy, flow cytometry analysis, western blot and migration assays were carried out to determine the anti-cancer mechanisms of PE.
  
  
  RESULTS
  MTT analysis showed that EE, PE and DCLM could inhibit cell proliferation on HepG2, SW480, MCF-7, K562 and HUVEC cell lines, especially PE fraction. HPLC analysis pointed out five main components which may contribute to the anti-proliferative activity of PE. Further study showed that PE increased LDH release, induced apoptosis, disrupted mitochondrial membrane potential and elevated intracellular reactive oxygen species (ROS) in HepG2 cells, whereas the antioxidant N-acetyl-l-cysteine (NAC) prevented PE-induced ROS generation. The results of western blot revealed that PE induced apoptosis in HepG2 cells by enhancing Bax/Bcl-2 ratio, increasing cytochrome c in cytosol and activating caspase-3/9. Meanwhile, high levels of ROS could induce DNA damage-mediated protein expression, AKT, ERK inactivation and SAPKs activation. Furthermore, PE conspicuously blocked the migration of HUVEC cells.
  
  
  CONCLUSIONS
  The present results demonstrated that PE induced apoptosis in HepG2 cells may be via a ROS-mediated signaling pathway.
authors.affiliation: Department of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China; Department of Pharmacy, General Hospital of Shenyang Military Area Command, Shenyang 110840, China.
authors.forename: Qiao
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: Department of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China.
authors.forename: Can
authors.initials: C
authors.lastname: Cui
authors.affiliation: Department of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China.
authors.forename: Cong-Qin
authors.initials: CQ
authors.lastname: Chen
authors.affiliation: Department of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China.
authors.forename: Xiao-Long
authors.initials: XL
authors.lastname: Hu
authors.affiliation: Department of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China.
authors.forename: Ya-Hui
authors.initials: YH
authors.lastname: Liu
authors.affiliation: Department of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China.
authors.forename: Yan-Hua
authors.initials: YH
authors.lastname: Fan
authors.affiliation: Department of Pharmacy, General Hospital of Shenyang Military Area Command, Shenyang 110840, China.
authors.forename: Wei-Hong
authors.initials: WH
authors.lastname: Meng
authors.affiliation: Department of Pharmacy, General Hospital of Shenyang Military Area Command, Shenyang 110840, China. Electronic address: zhaoqingchun1967@163.com.
authors.forename: Qing-Chun
authors.initials: QC
authors.lastname: Zhao
mesh_terms: D032429:Alpinia; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D049109:Cell Proliferation; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D056945:Hep G2 Cells; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D020014:K562 Cells; D061986:MCF-7 Cells; D010936:Plant Extracts / Q000302:isolation & purification / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects* / Q000502:physiology
chemical_list: D010936:Plant Extracts; D017382:Reactive Oxygen Species
keywords: Alpinia oxyplylla; Apoptosis; Chrysin (PubChem CID:5281607); Cytotoxicity; HepG2; ROS; Tectochrysin (PubChem CID:5281954); Yakuchinone A (PubChem CID:133145)