Upregulation of P-glycoprotein by probiotics in intestinal epithelial cells and in the dextran sulfate sodium model of colitis in mice

Abstract

. Upregulation of P-glycoprotein by probiotics in intestinal epithelial cells and in the dextran sulfate sodium model of colitis in mice. Am J Physiol Gastrointest Liver Physiol 300: G1115-G1123, 2011. First published February 24, 2011 doi:10.1152/ajpgi.00027.2011.-P-glycoprotein (P-gp) mediates efflux of xenobiotics and bacterial toxins from the intestinal mucosa into the lumen. Dysregulation of P-gp has been implicated in inflammatory bowel disease. Certain probiotics have been shown to be effective in treating inflammatory bowel disease. However, direct effects of probiotics on P-gp are not known. Current studies examined the effects of Lactobacilli on P-gp function and expression in intestinal epithelial cells. Caco-2 monolayers and a mouse model of dextran sulfate sodium-induced colitis were utilized. P-gp activity was measured as verapamil-sensitive [ 3 H]digoxin transepithelial flux. Multidrug resistant 1 (MDR1)/P-gp expression was measured by real-time quantitative PCR and immunoblotting. Culture supernatant (CS; 1:10 or 1:50, 24 h) of Lactobacillus acidophilus or Lactobacillus rhamnosus treatment of differentiated Caco-2 monolayers (21 days postplating) increased (ϳ3-fold) MDR1/P-gp mRNA and protein levels. L. acidophilus or L. rhamnosus CS stimulated P-gp activity (ϳ2-fold, P Ͻ 0.05) via phosphoinositide 3-kinase and ERK1/2 MAPK pathways. In mice, L. acidophilus or L. rhamnosus treatment (3 ϫ 10 9 colonyforming units) increased mdr1a/P-gp mRNA and protein expression in the ileum and colon (2-to 3-fold). In the dextran sulfate sodium (DSS)-induced colitis model (3% DSS in drinking water for 7 days), the degree of colitis as judged by histological damage and myeloperoxidase activity was reduced by L. acidophilus. L. acidophilus treatment to DSS-treated mice blocked the reduced expression of mdr1a/ P-gp mRNA and protein in the distal colon. These findings suggest that Lactobacilli or their soluble factors stimulate P-gp expression and function under normal and inflammatory conditions. These data provide insights into a novel mechanism involving P-gp upregulation in beneficial effects of probiotics in intestinal inflammatory disorders. multidrug resistance 1; Lactobacillus species; phosphoinositide 3-kinase; ERK1/2 kinase; dextran sulfate sodium-induced colitis; intestinal inflammation

Lactobacillus acidophilusstimulates intestinal P-glycoprotein expression via a c-Fos/c-Jun-dependent mechanism in intestinal epithelial cells

“…LA (strain 4357, American Type Culture Collection) was grown overnight, and CS was obtained as described previously (48). For our studies, LA CS was diluted in a ratio of 1:10 in cell culture medium supplemented with 1% FBS.…”

“…Caco-2 cells were grown routinely in 75-cm 2 plastic flasks in minimum essential medium (pH 7.2) supplemented with high-glucose 20% FBS, 20 mM HEPES, 100 IU/ml penicillin, and 100 g/ml streptomycin in 5% CO 2-95% O2 at 37°C (48,49). Cells at passages 25-45 were used for these studies.…”

“…Our previous studies showed that LA gavage demonstrated an increase in Pgp expression in the ileum and colon and attenuated decreased Pgp expression in the colon of DSS-colitis mice (48), suggesting that the effects of LA CS on intestinal Pgp may have clinical significance. These studies also demonstrated that LA CS-induced upregulation of Pgp in IECs occurred via a transcriptional mechanism.…”

“…However, the exact mechanisms by which an individual probiotic strain exerts its effect on intestinal epithelial functions during intestinal inflammation are not well understood. We previously showed that the culture supernatant (CS) of Lactobacillus acidophilus (LA) significantly stimulated function, expression, and promoter activity of the efflux transporter P-glycoprotein (Pgp) (48).…”

Reduction of MDR1, MRP1 and BCRP expression in Crohn’s disease: a role in disease pathogenesis?

“…REG3G prevents exaggerated inflammation by bactericidal activity and suppression of cytokine production (33). The interplay of MDR1 and REG3G in the balance of intestinal homeostasis is still unknown, but both are regulated by microbial colonization (34). Downregulated MDR1 expression in the ileum is in conflict with increased REG3G expression in CD patients by Gironella et all (33) and assume different regulatory mechanisms.…”