ABSTRACT
Objective:To observe the protective effects of Clostridium butyricum and butyrate on pancreas, liver and intestinal mucosa in rats with acute necrotizing pancreatitis (ANP), and to explore the possible mechanism.Methods:Forty Sprage-dawley rats were randomly divided into control group, ANP group，Clostridium butyricum treated group(CB group) and butyrate treated group(SB group), with 10 rats in each group by random number method. The ANP rat models were prepared by retrograde injection of sodium taurocholate into the biliopancreatic duct. Rats in CB and SB group were given intragastic administration of Clostridium butyricum 1×10 9 CFU or sodium butyrate (100 mg/kg) in 10 days once a day before modeling. Serum amylase (SAMY), lypase, ALT, AST, TBil, tumor necrosis factor alpha(TNF-α), IL-6 and HMGB1were measured after 24 h. Protein from intestinal mucosa was extracted and Western Blotting was used to measure expression of tight-junction proteins ZO-1, claudin-1 and occludin. Pancreas and liver tissues were stained with hematoxylin-eosin and scored by pathology. Results:The levels of amylase [(9365.1±716.5), (5947.3±512.0), (6517.7±269.6)U/L], lipase[(8343.7±1041.4), (6600.4±899.7), (6754.4±1046.4)U/L], AST[(560.5±72.7), (432.0±76.2), (429.8±40.5)U/L], ALT[(499.9±65.2), (385.7±46.0), (395.8±45.8)U/L], TBil[(134.2±56.2), (74.3±65.2), (81.3±35.3)U/L], TNF-α[(162.0±14.4), (100.4±6.3), (119.2±12.5)ng/L], IL-6[(161.4±26.0), (104.8±15.2), (105.5±12.7)ng/L], HMGB1[(100.1±6.7), (58.0±7.7), (63.4±7.2)ng/L] in ANP group, CB group and SB group were detected; and the pathological scores of pancreas[(11.2±1.08)，(9.45±1.06), (9.04±0.89)] and liver[(2.89±0.73), (2.09±0.49), (2.12±0.52)] in ANP group，CB group and SB group were higher than those in control group[(100.6±5.20)U/L, (966.5±301.9)U/L，(30.2±6.3)U/L, (27.6±5.9)U/L, (2.4±0.6)U/L, (29.5±4.8)ng/L，(36.9±7.6)ng/L，(35.5±5.7)ng/L，(1.18±0.05)，(0.56±0.09)]. However, those indexes in CB group and SB group were lower than those in ANP group, and the difference was statistically significant (all P<0.05). The expression of ZO-1 in control group, ANP group, CB group and SB group was 1.83, 0.79, 1.25 and 1.16. The expression of claudin-1 in control group, ANP group, CB group and SB group was 0.58, 0.13, 0.43 and 0.37. The expression of occludin in control group, ANP group, CB group and SB group was 1.06, 0.38, 0.82 and 0.79. The expression of TJ proteins in ANP group was significantly lower than that in other groups and the difference was statistically significant (all P<0.05). Conclusions:Clostridium butyricum and metabolites butyrate can alleviate the inflammatory response in ANP rats with liver injury, maintain the function of intestinal mucosal barrier and prevent the liver injury.
ABSTRACT
Objective To explore the effect of Clostridium butyricum ( C. butyricum ) and its metabolite butyrate on the function of intestinal mucosal barrier and intestinal flora in acute necrotizing pancreatitis ( ANP) rats with intra-abdominal hypertension ( IAH) . Methods Eighty SD rats were randomly divided into normal control group (A group, n=20), ANP with IAH group(B group, n=20), ANP with IAH and C. butyricum treated group ( C group, n=20 ) , ANP with IAH and sodium butyrate treated group ( D group, n=20). Rats of C and D group were given intragastric administration of C. butyricum 1 × 109 CFU once a day or 100 mg/kg sodium butyrate once a day from 10 days before modeling. Sodium taurocholate injection method via pancreatobiliary ducts was used to establish ANP with IAH rat model, and the intra-abdominal pressure was measured by direct puncture of left lower belly 24 h after modeling. Blood samples were collected for detecting serum amylase(AMY), tumor necrosis factor alpha (TNF-α), diamine oxidase( DAO ) , lipopolysaccharide ( LPS ) and D-Lactate, and the pathological changes of terminal ileum was observed. Real-time quantitative PCR was used to detect the populations of 6 bacteria in ileum mucosa. Results The levels of AMY, TNF-α, LPS,DAO, D-Lactate and ileum mucosa score were obviously higher in B, C and D group than those in A group, but the number of piobiotic flora in ileum mucosa was lower than that in A group, while the number of pathogenic bacteria was higher than that in A group. The levels of LPS, DAO, D-Lactate and ileum mucosa pathological score were lower in C group and D group than those in B group, but the number of piobiotic flora in ileum mucosa was lower than that in B group, while the number of pathogenic bacteria was higher than that in B group. All the differences above were statistically different (P<0.05). Conclusions C. butyricum and butyrate can maintain the function of intestinal mucosal barrier in ANP rats with IAH, and also readjust the imbalance of intestinal flora.