title: Cordyceps militaris extract stimulates Cl(-) secretion across human bronchial epithelia by both Ca(2+)(-) and cAMP-dependent pathways.
abstract: BACKGROUND
  The caterpillar fungus Cordyceps militaris (CM; Clavicipitaceae) is a well-known traditional Chinese medicine that can be artificially cultivated on a large scale. We have previously demonstrated that its stimulatory action on ion transport in human airway epithelia is similar to Cordyceps sinensis (Clavicipitaceae), which has been traditionally used to treat respiratory diseases.
  
  
  OBJECTIVE
  To investigate the signal transduction mechanism(s) underlying CM-induced ion transport activity in cultured human bronchial epithelia.
  
  
  METHODS
  16HBE14o-, a human bronchial epithelial cell line, was used to study the regulation of ion transport by the water extract of CM. CM extract was added to the apical or basolateral aspect of the epithelia. In subsequent experiments, different Cl(-) channel and K(+) channel blockers, adenylate cyclase and protein kinase A (PKA) inhibitors, and an intracellular Ca(2+) chelator were used to examine the involvement of apical Cl(-) and basolateral K(+) channels in mediating CM-induced Cl(-) secretion and the underlying signal transduction mechanism(s). PKA activity was also measured in 16HBE14o- cells.
  
  
  RESULTS
  CM stimulated Cl(-) secretion across 16HBE14o- monolayers in a dose-dependent manner. Cl(-) secretion could be inhibited by apical application of the cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-)channel blocker and the calcium-activated Cl(-) channel (CaCC) blocker. Cl(-) secretion was sensitive to basolateral application of different K(+) channel blockers. Similar inhibitory patterns were obtained in nystatin-permeabilized epithelia. The CM-induced Cl(-) secretion could be inhibited by adenylate cyclase and PKA inhibitors as well as an intracellular Ca(2+) chelator. Data from the PKA assay suggested that CM extract caused a significant increase in PKA activity compared with untreated control epithelia.
  
  
  CONCLUSIONS
  These results suggest that CM extract stimulated Cl(-) secretion across human bronchial epithelia, possibly via apical CFTR and CaCC, and the basolateral K(+) channels are involved in driving apical Cl(-) exit. The underlying signal transduction mechanisms involve both cAMP- and Ca(2+)-dependent pathways.
authors.affiliation: School of Biomedical Sciences, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong.
authors.forename: Jacky Chun-Kit
authors.initials: JC
authors.lastname: Fung
authors.forename: Grace Gar-Lee
authors.initials: GG
authors.lastname: Yue
authors.forename: Kwok-Pui
authors.initials: KP
authors.lastname: Fung
authors.forename: Xin
authors.initials: X
authors.lastname: Ma
authors.forename: Xiao-Qiang
authors.initials: XQ
authors.lastname: Yao
authors.forename: Wing-Hung
authors.initials: WH
authors.lastname: Ko
mesh_terms: D000262:Adenylyl Cyclases / Q000378:metabolism; D001688:Biological Products / Q000494:pharmacology; D001980:Bronchi / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D002118:Calcium / Q000378:metabolism*; D002478:Cells, Cultured; D018118:Chloride Channels / Q000187:drug effects* / Q000378:metabolism; D002712:Chlorides / Q000378:metabolism*; D032661:Cordyceps; D000242:Cyclic AMP / Q000378:metabolism*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D004848:Epithelium / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D017136:Ion Transport / Q000187:drug effects*; D026902:Potassium Channel Blockers / Q000494:pharmacology; D020545:Respiratory Mucosa / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D001688:Biological Products; D018118:Chloride Channels; D002712:Chlorides; D026902:Potassium Channel Blockers; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; D000262:Adenylyl Cyclases; D002118:Calcium