Probiotics prevent intestinal barrier dysfunction in acute pancreatitis in rats via induction of ileal mucosal glutathione biosynthesis.
BACKGROUND
During acute pancreatitis (AP), oxidative stress contributes to intestinal barrier failure. We studied actions of multispecies probiotics on barrier dysfunction and oxidative stress in experimental AP.


RESULTS
Fifty-three male Spraque-Dawley rats were randomly allocated into five groups: 1) controls, non-operated, 2) sham-operated, 3) AP, 4) AP and probiotics and 5) AP and placebo. AP was induced by intraductal glycodeoxycholate infusion and intravenous cerulein (6 h). Daily probiotics or placebo were administered intragastrically, starting five days prior to AP. After cerulein infusion, ileal mucosa was collected for measurements of E. coli K12 and (51)Cr-EDTA passage in Ussing chambers. Tight junction proteins were investigated by confocal immunofluorescence imaging. Ileal mucosal apoptosis, lipid peroxidation, and glutathione levels were determined and glutamate-cysteine-ligase activity and expression were quantified. AP-induced barrier dysfunction was characterized by epithelial cell apoptosis and alterations of tight junction proteins (i.e. disruption of occludin and claudin-1 and up-regulation of claudin-2) and correlated with lipid peroxidation (r>0.8). Probiotic pre-treatment diminished the AP-induced increase in E. coli passage (probiotics 57.4+/-33.5 vs. placebo 223.7+/-93.7 a.u.; P<0.001), (51)Cr-EDTA flux (16.7+/-10.1 vs. 32.1+/-10.0 cm/s10(-6); P<0.005), apoptosis, lipid peroxidation (0.42+/-0.13 vs. 1.62+/-0.53 pmol MDA/mg protein; P<0.001), and prevented tight junction protein disruption. AP-induced decline in glutathione was not only prevented (14.33+/-1.47 vs. 8.82+/-1.30 nmol/mg protein, P<0.001), but probiotics even increased mucosal glutathione compared with sham rats (14.33+/-1.47 vs. 10.70+/-1.74 nmol/mg protein, P<0.001). Glutamate-cysteine-ligase activity, which is rate-limiting in glutathione biosynthesis, was enhanced in probiotic pre-treated animals (probiotics 2.88+/-1.21 vs. placebo 1.94+/-0.55 nmol/min/mg protein; P<0.05) coinciding with an increase in mRNA expression of glutamate-cysteine-ligase catalytic (GCLc) and modifier (GCLm) subunits.


CONCLUSIONS
Probiotic pre-treatment diminished AP-induced intestinal barrier dysfunction and prevented oxidative stress via mechanisms mainly involving mucosal glutathione biosynthesis.
Lutgendorff
Femke
F
Department of Clinical and Experimental Medicine, Division of Surgery, Linköping University, Linköping, Sweden.
Nijmeijer
Rian M
RM
Sandström
Per A
PA
Trulsson
Lena M
LM
Magnusson
Karl-Eric
KE
Timmerman
Harro M
HM
van Minnen
L Paul
LP
Rijkers
Ger T
GT
Gooszen
Hein G
HG
Akkermans
Louis M A
LM
Söderholm
Johan D
JD
D000818:Animals; D017209:Apoptosis; D004847:Epithelial Cells / Q000473:pathology; D005978:Glutathione / Q000096:biosynthesis*; D007082:Ileum; D007408:Intestinal Absorption; D007413:Intestinal Mucosa / Q000378:metabolism*; D015227:Lipid Peroxidation; D008297:Male; D018384:Oxidative Stress; D010195:Pancreatitis / Q000139:chemically induced / Q000628:therapy*; D019936:Probiotics / Q000494:pharmacology / Q000627:therapeutic use*; D051381:Rats; D017207:Rats, Sprague-Dawley; D019108:Tight Junctions / Q000737:chemistry; D015533:Transcriptional Activation; D016896:Treatment Outcome
D005978:Glutathione