Hesperidin stimulates cystic fibrosis transmembrane conductance regulator-mediated chloride secretion and ciliary beat frequency in sinonasal epithelium.
OBJECTIVE
Pharmacologic agents designed to promote mucociliary clearance (MCC) in chronic rhinosinusitis (CRS) represent a novel therapeutic strategy. The objectives of the present study were to investigate whether the natural bioflavonoid hesperidin 1) increases transepithelial chloride (Cl(-)) secretion in vitro and in vivo, 2) enhances ciliary beat frequency (CBF), and 3) exerts its mechanistic effects through cAMP/PKA-dependent pathways.


METHODS
In vitro and in vivo study.


METHODS
Laboratory.


METHODS
Transepithelial Cl(-) transport (Ussing chamber) and CBF were investigated in primary murine nasal septal (MNSE) and human sinonasal epithelial (HSNE) cultures. In vivo activity was measured using the murine nasal potential difference (NPD) assay, cystic fibrosis transmembrane conductance regulator (CFTR) R-domain phosphorylation, and cAMP levels were investigated to rule out a cAMP/PKA-dependent mechanism of activation.


RESULTS
Hesperidin significantly increased CFTR-mediated Cl(-) transport (change in short-circuit current, DeltaI(SC)) in both MNSE (13.51 +/- 0.77 vs 4.4 +/- 0.66 [control]; P < 0.05) and HSNE (12.28 +/- 1.08 vs 0.69 +/- 0.32 [control]; P < 0.05). Cl(-) transport across in vivo murine nasal epithelium was also significantly enhanced with hesperidin (-2.3 +/- 1.0 vs -0.8 +/- 0.8 mV [control], P < 0.05). There was no increase in cellular cAMP or phosphorylation of the CFTR R-domain. Hesperidin significantly increased CBF (ratio of pretreatment to post-treatment) with both basal (1.31 +/- 0.07 vs 0.93 +/- 0.06 [control]; P < 0.05), apical (1.72 +/- 0.09 vs 1.40 +/- 0.07 [control]; P < 0.05), and basal + apical delivery (2.26 +/- 0.18 vs 1.60 +/- 0.21, respectively; P < 0.05).


CONCLUSIONS
Our in vitro and in vivo investigations provide strong support for future testing of this robust Cl(-) secretagogue and CBF activator in human clinical trials for CRS.
Azbell
Christopher
C
Department of Surgery/Division of Otolaryngology, University of Alabama at Birmingham, Birmingham, AL, USA.
Zhang
Shaoyan
S
Skinner
Daniel
D
Fortenberry
James
J
Sorscher
Eric J
EJ
Woodworth
Bradford A
BA
D000818:Animals; D018929:Cell Culture Techniques; D002712:Chlorides / Q000378:metabolism*; D002923:Cilia / Q000187:drug effects / Q000502:physiology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000502:physiology; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000187:drug effects* / Q000378:metabolism; D006569:Hesperidin / Q000494:pharmacology*; D006801:Humans; D017136:Ion Transport / Q000187:drug effects; D051379:Mice; D008810:Mice, Inbred C57BL; D009079:Mucociliary Clearance / Q000187:drug effects*; D009297:Nasal Mucosa / Q000187:drug effects* / Q000378:metabolism / Q000503:physiopathology; D010256:Paranasal Sinuses / Q000187:drug effects* / Q000378:metabolism / Q000503:physiopathology
D002712:Chlorides; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D006569:Hesperidin; D017868:Cyclic AMP-Dependent Protein Kinases