title: Yakammaoto inhibited human coxsackievirus B4 (CVB4)-induced airway and renal tubular injuries by preventing viral attachment, internalization, and replication.
abstract: BACKGROUND
  Yakammaoto is a prescription of traditional Chinese medicine (TCM) containing nine ingredients, including Ephedra sinica, Pinellia ternate, Zingiber officinale, Tussilago farfara, Aster tataricus, Ziziphus jujube, Belamcanda chinensis, Asarum sieboldii, and Schisandra chinensis. Yakammaoto has been used against flu-like symptoms for more than two thousand years in China and Japan. Coxsackievirus B4 (CVB4) causes not only flu-like symptoms but life-threatening diseases, such as pneumonia, acute kidney injury, and so forth with severe morbidity and mortality. There is no effective therapeutic modality against CVB4 infection. It is unknown whether yakammaoto is effective against CVB4 infection. We tested the hypothesis that yakammaoto can effectively inhibit CVB4-induced plaque formation in human airway and renal tubular cell lines by preventing viral attachment, internalization, and replication.
  
  
  METHODS
  The fingerprint of yakammaoto was assessed by HPLC. Effects of yakammaoto on CVB4 infection were tested by plaque reduction assay, reverse transcription polymerase chain reaction (RT-PCR), and enzyme-linked immunosorbent assay (ELISA).
  
  
  RESULTS
  Yakammaoto dose-dependently inhibited CVB4-induced plaque formation in HK-2, A549, and HEp-2 cells (p<0.0001). Yakammaoto was both effective when supplemented prior to and after viral inoculation (p<0.0001) by preventing viral attachment (p<0.0001), internalization (p<0.0001), and replication (p<0.0001). Yakammaoto could decrease NGAL secretion before cytolysis to protect against viral injury.
  
  
  CONCLUSIONS
  Yakammaoto had antiviral activity against CVB4-induced cellular injuries in airway mucosa and renal tubular epithelia by preventing viral attachment, internalization, and replication. The current study provides a basic support of its potential use against CVB4-induced airway and concomitant renal injuries.
authors.affiliation: School of Pharmacy and Graduate Institute of Natural Products, College of Pharmacy, Kaohsiung Medical University, Taiwan.
authors.forename: Ming Hong
authors.initials: MH
authors.lastname: Yen
authors.affiliation: Division of Nephrology, Department of Internal Medicine, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Taiwan.
authors.forename: Jia Jung
authors.initials: JJ
authors.lastname: Lee
authors.affiliation: Department of Renal Care, College of Medicine, Kaohsiung Medical University, Taiwan.
authors.forename: Chia Feng
authors.initials: CF
authors.lastname: Yeh
authors.affiliation: Department of Renal Care, College of Medicine, Kaohsiung Medical University, Taiwan.
authors.forename: Kuo Chih
authors.initials: KC
authors.lastname: Wang
authors.affiliation: Department of Renal Care, College of Medicine, Kaohsiung Medical University, Taiwan.
authors.forename: Ya Wen
authors.initials: YW
authors.lastname: Chiang
authors.affiliation: Department of Microbiology, School of Medicine, College of Medicine, Kaohsiung Medical University, Taiwan.
authors.forename: Lien Chai
authors.initials: LC
authors.lastname: Chiang
authors.affiliation: Department of Renal Care, College of Medicine, Kaohsiung Medical University, Taiwan; Division of Gastroenterology, Department of Internal Medicine, Kaohsiung Medical University Hospital, Kaohsiung Medical University, 100 Shih-Chuan 1st Road, Kaohsiung 807, Taiwan. Electronic address: cjs@kmu.edu.tw.
authors.forename: Jung San
authors.initials: JS
authors.lastname: Chang
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology*; D002460:Cell Line; D045744:Cell Line, Tumor; D003384:Coxsackievirus Infections / Q000188:drug therapy; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D029822:Enterovirus B, Human / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D016899:Interferon-beta / Q000378:metabolism; D007684:Kidney Tubules / Q000166:cytology; D012137:Respiratory System / Q000166:cytology; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D053585:Virus Attachment / Q000187:drug effects; D053586:Virus Internalization / Q000187:drug effects; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000998:Antiviral Agents; D004365:Drugs, Chinese Herbal; D014409:Tumor Necrosis Factor-alpha; C000591854:yakammaoto; D016899:Interferon-beta
keywords: 3,3',5,5'-tetramethylbenzidine; 50% cytotoxic concentration; ATCC; CC(50); CVB4; DMEM; DMSO; Dulbecco's Modified Eagle medium; ELISA; FCS; HEp-2; HK-2; IC(50); IFN; Kidney injury; PBS; Respiratory tract infection; SI; She-Gan-Ma-Huang-Tang; Sheganmahuang decoction; Sheganmahuang decoction (Chinese); TCM; TMB; Yakammaoto; Ye-Gan-Ma-Huang-Tang; Ye-Gan-Ma-Huang-Tang (Taiwan); dimethylsulfoxide; enzyme-linked immunosorbent assay; fetal calf serum; human coxsackievirus B4; human kidney 2 cell; human larynx epidermoid carcinoma cell; interferon; minimal concentration required to inhibit 50% plaque formation; pfu; phosphate-buffered saline; plaque forming unit; selectivity index; the American Type Culture Collection; traditional Chinese medicine