title: Effects of Viola yedoensis Makino anti-itching compound on degranulation and cytokine generation in RBL-2H3 mast cells.
abstract: BACKGROUND
  The Chinese herb compound prescription Viola yedoensis Makino Anti-itching Compound (VYAC), which consists of Viola yedoensis Makino, herb, Sophora flavescens Aiton, root, and Dictamnus dasycarpus Turcz, root and rhizome, has been traditionally used to treat various skin allergic inflammatory diseases in clinic.
  
  
  OBJECTIVE
  The aim of this study is to investigate the effects of VYAC on degranulation and to determine its anti-inflammatory mechanism in RBL-2H3 mast cells.
  
  
  METHODS
  VYAC was extracted with water-coction extraction (Shufen et al., 2012). The aqueous extracts were concentrated in vacuum under reduced pressure and lyophilized using a freeze dryer, and lyophilized powder was obtained. MTT was used to evaluate the cytotoxic of VYAC on RBL-2H3 cells. Degranulation was carried out with RBL-2H3 cell model, which was stimulated with A23187 plus PMA. β-Hexosaminidase and histamine were measured to evaluate degranulation. The mRNA levels of inflammation cytokines (IL-1β, TNF-α, IL-6, and iNOS) were investigated by RT-PCR to explain the anti-inflammatory mechanism of VYAC.
  
  
  RESULTS
  VYAC did not show cytotoxic effect on RBL-2H3 cells in the range of 25-400μg/mL. A higher dose of VYAC (800μg/mL) showed significant cytotoxicity (P<0.05). VYAC could significantly inhibit β-hexosaminidase and histamine release when treated with 100, 200, and 400μg/mL (P<0.05), but could not significantly inhibit β-Hexosaminidase and histamine release when treated with 25 and 50μg/mL (p>0.05). The mRNA levels of inflammatory cytokines (TNF-α, IL-1β, IL-6, and iNOS) could significantly decrease when treated with 200 and 400μg/mL (P<0.05) of VYAC, which were associated with the development of inflammation.
  
  
  CONCLUSIONS
  Results showed that VYAC inhibited β-hexosaminidase and histamine release, which was inhibit A23187 plus PMA stimulated RBL-2H3 cell degranulation and downregulated inflammatory cytokines (IL-1β, TNF-α, IL-6, and iNOS) expression to block inflammatory development.
authors.affiliation: Experiment Center for Teaching and Learning, Shanghai University of Traditional Chinese Medicine, 1200 Cailun Rood, Pudong New District, Shanghai 201203, China; Murad Research Center for Moderniszed Chinese Medicine, Shanghai University of Traditional Chinese Medicine, 1200 Cailun Rood, Pudong New District, Shanghai 201203, China.
authors.forename: Hai-Rong
authors.initials: HR
authors.lastname: Zeng
authors.affiliation: Experiment Center for Teaching and Learning, Shanghai University of Traditional Chinese Medicine, 1200 Cailun Rood, Pudong New District, Shanghai 201203, China; School of Pharmacy, Shanghai University of Traditional Chinese Medicine, 1200, Cailun Road, Pudong New District, Shanghai 201203, China. Electronic address: annabel_cn@163.com.
authors.forename: Bing
authors.initials: B
authors.lastname: Wang
authors.affiliation: Experiment Center for Teaching and Learning, Shanghai University of Traditional Chinese Medicine, 1200 Cailun Rood, Pudong New District, Shanghai 201203, China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Zhao
authors.affiliation: Experiment Center for Teaching and Learning, Shanghai University of Traditional Chinese Medicine, 1200 Cailun Rood, Pudong New District, Shanghai 201203, China; School of Pharmacy, Shanghai University of Traditional Chinese Medicine, 1200, Cailun Road, Pudong New District, Shanghai 201203, China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: Experiment Center for Teaching and Learning, Shanghai University of Traditional Chinese Medicine, 1200 Cailun Rood, Pudong New District, Shanghai 201203, China.
authors.forename: Mei-Yun
authors.initials: MY
authors.lastname: Liang
authors.affiliation: Experiment Center for Teaching and Learning, Shanghai University of Traditional Chinese Medicine, 1200 Cailun Rood, Pudong New District, Shanghai 201203, China.
authors.forename: Ya-Qi
authors.initials: YQ
authors.lastname: Yao
authors.affiliation: Murad Research Center for Moderniszed Chinese Medicine, Shanghai University of Traditional Chinese Medicine, 1200 Cailun Rood, Pudong New District, Shanghai 201203, China; Department of Biochemistry and Molecular Biology, George Washington University of USA, WA 20052, USA.
authors.forename: Ka
authors.initials: K
authors.lastname: Bian
authors.affiliation: Experiment Center for Teaching and Learning, Shanghai University of Traditional Chinese Medicine, 1200 Cailun Rood, Pudong New District, Shanghai 201203, China. Electronic address: zwr885@hotmail.com.
authors.forename: Wei-Rong
authors.initials: WR
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D000982:Antipruritics / Q000494:pharmacology*; D015550:Cell Degranulation / Q000187:drug effects*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D016207:Cytokines / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D004365:Drugs, Chinese Herbal / Q000737:chemistry / Q000493:pharmacokinetics / Q000494:pharmacology*; D006632:Histamine / Q000276:immunology / Q000378:metabolism*; D018836:Inflammation Mediators / Q000276:immunology / Q000378:metabolism*; D008407:Mast Cells / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D052247:Nitric Oxide Synthase Type II / Q000235:genetics / Q000378:metabolism; D008517:Phytotherapy; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D032413:Viola / Q000737:chemistry*; D001619:beta-N-Acetylhexosaminidases / Q000378:metabolism
chemical_list: D000893:Anti-Inflammatory Agents; D000982:Antipruritics; D016207:Cytokines; D004365:Drugs, Chinese Herbal; D018836:Inflammation Mediators; D012333:RNA, Messenger; C000617254:qizhiweitong; D006632:Histamine; D052247:Nitric Oxide Synthase Type II; C496318:Nos2 protein, rat; D001619:beta-N-Acetylhexosaminidases
keywords: Anti-allergic; Anti-inflammation; Degranulation; Iola Yedoensis Makino anti-itching compound; RBL-2H3