Effects Of Probiotics Supplementation On Liver And Intestinal Mucosal Injury Induced By Pyrazinamide In Rats

Objective Pyrazinamide is an antituberculous drug,which can cause liver and intestinal mucosal barrier damage in patients with antituberculous therapy.Probiotics are a kind of living microorganisms which are beneficial to our body.The purpose of this study was to investigate the effects of probiotics(Lactobacillus casei)supplementation on hepatic and intestinal mucosal injury induced by pyrazinamide in rats,and to provide a reference basis for the rational application of probiotics to improve the adverse reactions of anti-tuberculosis drugs.Methods Forty adult male Sprague-Dawley rats were randomly divided into four groups: normal control group(C group),pyrazinamide group(L0 group),low dose Lc S group(L1 group)and high dose Lc S group(L2 group).Group Z,L1 and L2 were established model of liver injury induced by pyrazinamide,while L1 group and L2 group were given Lactobacillus casei(Lc S)(108 CFU·m L-1)intervention dose for a day of 10 m L·kg-1·d-1 and 20 m L·kg-1·d-1 by gavage,and the animal experiment last for 10 weeks.HE staining was used to observe the pathological changes of liver and colon in rats,and the histopathological scores of liver were evaluated;Rate method were used to detected the serum levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST);the expression of Occludin and ZO-1 were detected by Western blot;quantitative analysis of bifidobacteria,lactobacillus and escherichia coli 16 Sr DNA V3 variable in rat feces by real-time fluorescence quantitative polymerase chain reaction(RT-PCR).The levels of secretory immunoglobulin A(s Ig A),?-defensin-2(?D-2)in colon,and the levels of lipopolysaccharide(LPS),tumor necrosis factor-?(TNF-?),interleukin-6(IL-6),interleukin-10(IL-10)and interleukin-12(IL-12)in serum and colon were measured by enzyme linked immunosorbent assay.Results After 10 weeks of treatment with pyrazinamide,HE staining of L0 group showed that the liver cells appeared moderate edema,ballooning degeneration,with inflammatory cell infiltration,which pathological score reached 3.20.And the serum ALT and AST levels were significantly higher than those in group C,it's showed that liver injury was successful modeling.The depth of colon recess,the proportion of epithelial goblet cells,the expression level of Occludin and ZO-1 of colonic tight junction protein and the level of s Ig A were significantly decreased,while the level of ?D-2 was increased by 48.1%.The number of Escherichia coli in feces of rats increased significantly while the number of Bifidobacterium and Lactobacillus decreased significantly at the end of the 6th week of intervention,results showed that pyrazinamide caused damage to the intestinal mucosa of rats.The levels of LPS?TNF-??IL-6 and IL-12 in serum and colon of rats were higher than those in the control group(P < 0.05),and the levels of IL-10 in serum and colon were lower than those in the control group.After 10 weeks of the intervention with different doses of probiotic,the hepatic lobular structure of rats in L1 group and L2 group were significantly improved than those in the L0 group,pathological score was reduced to 0.40 points and 0.30 points,the serum of ALT and AST levels were reduced to 73.10 U/L,73.60 U/L and 150.50 U/L,143.80 U/L,significantly lower than that in the L0 group(95.90 U/L and 188.60 U/L),all these results suggest that supplementation of probiotics could improve liver injury.The depth of colonic recess increased by 41.5% and 59.7%,respectively.And the ?D-2 level significantly decreased while the level of s Ig A significantly increased.In the L2 group,the proportion of epithelial goblet cells increased by 7.6%,and the expression of Occludin and ZO-1 increased by 59.7% and 97.9%,respectively.At the end of the 6th week and the 10 th week of intervention,the levels of Bifidobacterium and Lactobacillus increased significantly.The amount of Escherichia coli decreased significantly.These suggest that probiotics can improve the intestinal mucosal barrier injury in rats.The serum levels of TNF-?,IL-6 and IL-12 in L1 group and L2 group were significantly lower than those in the L0 group,and the level of IL-10 reached 1.2 times than that in the L0 group.The levels of LPS,IL-6 and IL-12 in colon were all decreased while the level of IL-10 increased significantly.Compared with the L0 group,the serum LPS level in L2 group was decreased by 26.2%,and the level of TNF-? in colon was 55.4% lower.There was a correlation between LPS and IL-6 in serum.Conclusion Pyrazinamide can cause liver,intestinal flora and intestinal mucosal injury in rats.Probiotics can improve intestinal barrier injury by regulating intestinal microecological balance,promoting tight junction protein expression and enhancing intestinal mucosal immunity.And through the "hepatointestinal axis",the further improvement of liver injury has a certain effect.