title: Anti-allergic potential of Typhonium blumei: Inhibition of degranulation via suppression of PI3K/PLCγ2 phosphorylation and calcium influx.
abstract: BACKGROUND
  Typhonium blumei Nicolson & Sivadasan (Araceae) is a traditional Chinese medicinal herb possessing detumescent, detoxifying, and anti-inflammatory activities. It is used in Taiwan as a folk medicine to treat cancer and inflammatory diseases. Typhonium blumei is usually not distinguished from Typhonium roxburghii Schott and they are commonly used interchangeably.
  
  
  OBJECTIVE
  To evaluate and compare the anti-allergic and anti-inflammatory properties of T. blumei and T. roxburghii, their composition profiles and molecular basis of the anti-allergic effect.
  
  
  METHODS
  The methanolic plant extracts were partitioned with different solvents to obtain the nonpolar fractions. The anti-allergic activity of the nonpolar fractions was assessed by A23187- and antigen-induced degranulation assays using RBL-2H3 mast cells. Several molecular targets were investigated: FcεRI receptor expression by flow cytometry, calcium influx by live cells imaging fluorescent microscopy, cytokines mRNA expression by RT-PCR, and protein expression by Western blotting. The anti-inflammatory activity was evaluated using superoxide anion and elastase release assays in human neutrophils. TLC, NMR and GC-MS analyses were conducted to evaluate the chemical composition of the fractions.
  
  
  RESULTS
  The nonpolar fractions of both Typhonium species showed potent inhibitory activity in A23187-induced degranulation assay in RBL-2H3 cells. They also inhibited superoxide production and elastase release in human neutrophils. T. blumei nonpolar fractions inhibited antigen-induced β-hexosaminidase and histamine release. The nonpolar fractions of T. blumei significantly inhibited calcium influx upon activation with either A23187 or an antigen. The fractions did not affect FcεRI receptor expression, mRNA level of IL-4 and MCP-1 cytokine production or MAPK proteins expression, but did suppress the calcium signaling pathway via PI3K/PLCγ2. The active fractions were rich in fatty acids with palmitic, linoleic and α-linolenic acids identified as the major fatty acids in both plants. The content of omega-3 unsaturated fatty acids was higher in T. roxburghii nonpolar fractions compared to T. blumei.
  
  
  CONCLUSIONS
  Both species possess potent anti-allergic and anti-inflammatory activities. The inhibition of degranulation in mast cells was attributed to calcium influx modulation. The obtained results support the traditional use of T. blumei in the treatment of inflammatory diseases as well as its substitution with T. roxburghii.
authors.affiliation: Graduate Institute of Natural Products, College of Pharmacy, Kaohsiung Medical University, Kaohsiung 807, Taiwan; Department of Biotechnology, College of Life Science, Kaohsiung Medical University, Kaohsiung 807, Taiwan. Electronic address: mickorinek@hotmail.com.
authors.forename: Michal
authors.initials: M
authors.lastname: Korinek
authors.affiliation: Department of Biotechnology, College of Life Science, Kaohsiung Medical University, Kaohsiung 807, Taiwan. Electronic address: m0313ing@hotmail.com.
authors.forename: Kuan-Ming
authors.initials: KM
authors.lastname: Chen
authors.affiliation: Department of Biotechnology, College of Life Science, Kaohsiung Medical University, Kaohsiung 807, Taiwan. Electronic address: yhjiang0508@gmail.com.
authors.forename: Yu-Han
authors.initials: YH
authors.lastname: Jiang
authors.affiliation: Graduate Institute of Natural Products, College of Pharmacy, Kaohsiung Medical University, Kaohsiung 807, Taiwan; Department of Pharmacognosy and Natural Products Chemistry, Faculty of Pharmacy, Ain-Shams University, Cairo 11566, Egypt. Electronic address: elshazly444@googlemail.com.
authors.forename: Mohamed
authors.initials: M
authors.lastname: El-Shazly
authors.affiliation: Education Center for Humanities and Social Sciences, National Yang Ming University, Taipei 112, Taiwan. Electronic address: inyime@gmail.com.
authors.forename: Joel
authors.initials: J
authors.lastname: Stocker
authors.affiliation: Graduate Institute of Natural Products, College of Pharmacy, Kaohsiung Medical University, Kaohsiung 807, Taiwan; Department of Biotechnology, College of Life Science, Kaohsiung Medical University, Kaohsiung 807, Taiwan. Electronic address: fatchou1988@hotmail.com.
authors.forename: Chon-Kit
authors.initials: CK
authors.lastname: Chou
authors.affiliation: Graduate Institute of Natural Products, College of Medicine, Chang Gung University, Taoyuan 333, Taiwan; Research Center for Industry of Human Ecology and Graduate Institute of Health Industry Technology, Chang Gung University of Science and Technology, Taoyuan 333, Taiwan; Department of Anesthesiology, Chang Gung Memorial Hospital, Taoyuan 333, Taiwan. Electronic address: htl@mail.cgu.edu.tw.
authors.forename: Tsong-Long
authors.initials: TL
authors.lastname: Hwang
authors.affiliation: Graduate Institute of Natural Products, College of Pharmacy, Kaohsiung Medical University, Kaohsiung 807, Taiwan; School of Pharmacy, College of Pharmacy, China Medical University, Taichung 404, Taiwan; Chinese Medicine Research and Development Center, China Medical University Hospital, Taichung 404, Taiwan; Center for Molecular Medicine, China Medical University Hospital, Taichung 404, Taiwan. Electronic address: yachwu@mail.cmu.edu.tw.
authors.forename: Yang-Chang
authors.initials: YC
authors.lastname: Wu
authors.affiliation: Department of Biotechnology, College of Life Science, Kaohsiung Medical University, Kaohsiung 807, Taiwan; The Institute of Biomedical Sciences, National Sun Yat-Sen University, Kaohsiung 804, Taiwan. Electronic address: bhchen@kmu.edu.tw.
authors.forename: Bing-Hung
authors.initials: BH
authors.lastname: Chen
authors.affiliation: Graduate Institute of Natural Products, College of Pharmacy, Kaohsiung Medical University, Kaohsiung 807, Taiwan; Research Center for Environmental Medicine, Kaohsiung Medical University, Kaohsiung 807, Taiwan; Cancer Center, Kaohsiung Medical University Hospital, Kaohsiung 807, Taiwan; Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung 804, Taiwan. Electronic address: aaronfrc@kmu.edu.tw.
authors.forename: Fang-Rong
authors.initials: FR
authors.lastname: Chang
mesh_terms: D000818:Animals; D018926:Anti-Allergic Agents / Q000494:pharmacology* / Q000627:therapeutic use; D000893:Anti-Inflammatory Agents / Q000494:pharmacology* / Q000627:therapeutic use; D029064:Araceae / Q000737:chemistry*; D002118:Calcium / Q000378:metabolism*; D015550:Cell Degranulation / Q000187:drug effects*; D002460:Cell Line; D016207:Cytokines / Q000378:metabolism; D004365:Drugs, Chinese Herbal / Q000494:pharmacology* / Q000627:therapeutic use; D005227:Fatty Acids / Q000032:analysis / Q000494:pharmacology* / Q000627:therapeutic use; D006636:Histamine Release / Q000187:drug effects; D006801:Humans; D015847:Interleukin-4 / Q000378:metabolism; D008407:Mast Cells / Q000187:drug effects / Q000378:metabolism; D009504:Neutrophils / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D051966:Phospholipase C gamma / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D008517:Phytotherapy; D051381:Rats; D017455:Receptors, IgE / Q000378:metabolism; D015398:Signal Transduction; D001619:beta-N-Acetylhexosaminidases / Q000378:metabolism
chemical_list: D018926:Anti-Allergic Agents; D000893:Anti-Inflammatory Agents; D016207:Cytokines; D004365:Drugs, Chinese Herbal; D005227:Fatty Acids; D017455:Receptors, IgE; D015847:Interleukin-4; D019869:Phosphatidylinositol 3-Kinases; D051966:Phospholipase C gamma; D001619:beta-N-Acetylhexosaminidases; D002118:Calcium
keywords: Anti-allergic; Anti-inflammatory; Calcium influx; Fatty acids; Typhonium blumei; Typhonium roxburghii