Effect of genistein on basal jejunal chloride secretion in R117H CF mice is sex and route specific

Abstract

Esa Rayyan,1 Sarah Polito,1 Lana Leung,1 Ashesh Bhakta,1 Jonathan Kang,1 Justin Willey,1 Wasim Mansour,1 Mitchell L Drumm,2 Layla Al-Nakkash11Department of Physiology, Arizona College of Osteopathic Medicine, Midwestern University, Glendale, AZ, USA; 2Pediatric Pulmonology Division, Case Western Reserve University, Cleveland, OH, USAAbstract: Cystic fibrosis (CF) results from the loss or reduction in function of the CFTR (cystic fibrosis transmembrane conductance regulatory protein) chloride channel. The third most common CFTR mutation seen clinically is R117H. Genistein, a naturally occurring phytoestrogen, is known to stimulate CFTR function in vitro. We aimed to determine whether route of administration of genistein could mediate differential effects in R117H male and female CF mice. Mice were fed (4 weeks) or injected subcutaneously (1 week) with the following: genistein 600 mg/kg diet (600Gd); genistein-free diet (0Gd); genistein injection 600 mg/kg body weight (600Gi); dimethyl sulfoxide control (0Gi). In male R117H mice fed 600Gd, basal short circuit current (Isc) was unchanged. In 600Gd-fed female mice, there was a subgroup that demonstrated a significant increase in basal Isc (53.14±7.92 µA/cm2, n=6, P<0.05) and a subgroup of nonresponders (12.05±6.59 µA/cm2, n=4), compared to 0Gd controls (29.3±6.5 µA/cm2, n=7). In R117H mice injected with 600Gi, basal Isc was unchanged in both male and female mice compared to 0Gi controls. Isc was measured in response to the following: the adenylate cyclase activator forskolin (10 µM, bilateral), bumetanide (100 µM, basolateral) to indicate the Cl- secretory component, and acetazolamide (100 µM, bilateral) to indicate the HCO3- secretory component; however, there was no effect of genistein (diet or injection) on any of these parameters. Jejunal morphology (ie, villi length, number of goblet cells per villus, crypt depth, and number of goblet cells per crypt) in R117H mice suggested no genistein-mediated difference among the groups. Serum levels of genistein were significantly elevated, compared to respective controls, by either 600Gd (equally elevated in males and females) or 600Gi (elevated more in females versus males). These data suggest a sex-dependent increase in basal Isc of R117H mice and that the increase is also specific for route of administration.Keywords: genistein, intestine, secretion, CFTR, R117H