Prevention And Cure Effects Of Lactobacillus On Hyperuricemia Induced By High Fructose Diet And Its Mechanism

Objective:To explore the effects of Lactobacillus brevis DM9505 on high fructose(HF)-induced hyperuricemia model mice,and lay a foundation for the development and utilization of probiotics.Methods:1 Grouping and modelingIn this experiment,Balb/c male mice were randomly divided into normal Control group(Control),high fructose model group(HF)and Probiotic intervention group(Probiotic),15 rats in each group.The mice in the model group and the probiotic group were given free drinking of 15%(w/v)high fructose water.The probiotic group was given Lactobacillus brevis DM9505 orally(1×10~9cfu/kg,0.2ml/day)treatment at the same time,the normal control group given tap water for free drinking.2 To investigate the effect of DM9505 on the fructose metabolism mechanism of uric acid production in high fructose fed miceThe diet and activity status of each group of mice were observed daily.The body weight and blood glucose of each group were measured every week.All the mice were killed after 8 weeks.Serum uric acid(UA)and lipids were measured by enzyme kit in all groups.The liver tissues of mice were biopsy and HE stained to observe the pathological changes.The liver tissues were homogenized,and the changes of AdenosineTriphosphate(ATP),AdenosineMonophosphate(AMP),Inosine,Transforming growth factorβ1(TGF-β1)and Reactive oxygen species(ROS)levels in liver tissues were detected by ELISA,and the level of fecal Inosine in mice was detected.The expression levels of NOX4mRNA in the liver tissues of mice were detected by RT-qPCR.3 To investigate the effect of DM9505 on the regulation of uric acid production in high fructose fed miceThe liver tissues were homogenized and the changes of Lipopolysaccharide(LPS),Interferonα(IFN-α),Xanthine oxidase(XOD)concentration and XOD activity in liver tissues of each group were detected by ELISA.The changes of XOD mRNA expression in liver tissues of each group were detected by RT-qPCR.4 To investigate the regulatory effect of DM9505 on the intestinal flora of hyperuricemia mice induced by high fructoseThe colonic contents DNA of each group was extracted,and the characteristics of intestinal microflora in mice were identified by PCR-DGGE and gel-cutting sequencing.The Illuminate Hiseq PE250 16S rDNA sequencing analysis was used to analyze the species diversity,taxonomic diversity,Beta diversity,Alpha diversity and CCA among the different groups.Results:1 Body weight and blood glucose in each group continued to rise one week after the experiment.After 8 weeks of experiment,the body weight and blood glucose were significantly increased between the model group and the normal control group(P<0.05),meanwhile the basic indexes of UA,TG and TC were also significantly increased(P<0.05).After the intervention of probiotics,the UA level of mice was significantly reduced(P<0.05),while other indicators were not significantly improved.Compared with normal control group,the expression of AMP,Inosine,TGF-β1,NOX4 mRNA,ROS and the fecal-Inosine in high fructose model group were significantly increased(P<0.05)),while ATP levels were significantly decreased(P<0.05);After the intervention of probiotics,the expression levels of all other substances in mice were decreased with different levels(P<0.05),except ATP and AMP contents did not change significantly.The HE staining of liver tissue in each group showed that compared with the normal control group,the liver cells in the high-fructose model group swelled and swollen,while the morphological changes of the liver cells in the probiotic group were improved.2 Compared with the normal control group,the expression of LPS,IFN-α,XOD concentration,XOD activity and XOD mRNA in the liver of high-fructose model group were significantly increased(P<0.05).After the intervention of probiotics,the levels of LPS,IFN-α,XOD concentration,XOD activity and XOD mRNA expression were significantly decreased(P<0.05).3 PCR-DGGE gel imaging maps showed that high fructose intestinal flora and the normal control group of mice model of mice intestinal flora there is obvious difference between the number and structure:there are many differences between the two groups of banding,and stripe number of bacteria in the intestine of mice model mice decreased significantly compared with normal group.The results showed that the pathogenic bacteria increased in the model group and the beneficial bacteria were increased.Gel-imaging of PCR-DGGE showed that there was a significant difference in the number and structure of intestinal microflora between the high-fructose model group and the normal control group:there were many differences between the two groups of banding,and the model group mice gut flora number significantly reduced compared with the normal group of mice.Banding gel sequencing results showed that the model group of mice increased pathogenic bacteria,while probiotic bacteria increased in the probiotics group.The 16S rDNA high-throughput sequencing results of the colon contents of mice in each group were shown.Compared with the normal control group,the number of species and the flora diversity of the model group were significantly lower.The UPGMA clustering tree showed that the probiotics intervention group was closer to the normal control group and the similarity was greater.The model group was far from the normal group and the probiotics intervention group,and the similarity was lower.Compared with the normal control group,the abundance of Bacteroidetes was decreased while the Proteobacteria,Eubacterium,Desulfovibrio and Enterococcus were increased in the model group.The above results showed that the structure of gut microbiota in high fructose model mice were changed,the normal flora decreased and the number of pathogenic bacteria increased.Compared with the model group,the abundance of Bacteroidetes,Anaerotruncus were significantly increased in the probiotic intervention group,while the Firmicutes,Saccharibacteria(Proteobacteria)significantly reduced.The above results showed that in probiotics intervention mice intestinal normal flora has increased,while the conditions of pathogenic bacteria were decreased.Conclusion:1 Lactobacillus brevis DM9505 not only can adjust the level of fructose metabolism intermediate-Inosine but also can affect the concentration of LPS and IFN-αto stimulate the expression of XOD and its concentration and activity,which can improve the hyperuricemia of mice induced by high-fructose diet.2 Lactobacillus DM9505 can modify the intestinal flora disorders in high-fructose induced mice to a certain extent,in particular,can make the normal flora increased,reduce the number of opportunistic pathogens.