Establishment Of Mice Model For Dysbiosis Of Intestinal Flora And The Influence To Intestinal Toll-like Receptors

Objective: 1.Utilize antibiotics to effect intestinal microbiology of Balb/c mice, to establish mice model for dysbiosis of intestinal microflora based on intestinal predominant microbial population changes; 2.Toll-like receptor-mediated signaling of intestinal epithelium cells(IECs) influenced by alteration of intestinal flora, to investigate the molecular interaction of commensal bacteria with the intestinal epithelium and the mucosal immune system;to supply infectious diseases therapy with new thinking and strategy.Methods:1.We administer Balb/c mice with different doses of ceftriaxone sodium, establishing mice model for dysbiosis of Intestinal flora. Utilize 5g/kg/d doses, 8 days to establish light micro dysbiosis; meanwhile utilize 8g/kg/d doses, 8 days to establish heavy micro dysbiosis.2.In normal group, light micro dysbiosis and heavy micro dysbiosis animals intestinal epithelium cells were isolated Using an EDTA-viration method; Toll-like recptor 2, 4 and 9 mRNA were determined by RT-PCR; NF-κBp65 was determined by RT-PCR and immunohistochemistry.Results:1.Establish consummate analytical method of fourteen kinds of intestinal dominant microbe including Bacillus bifidus, Lactobacillus, Bateroides, Eubacterium, Enterococus, Streptococcus, Fusobacterium, Veillonella, Escherichia, Staphylococcus, yeast and so on.2.Using 5g/kg/d doses of ceftriaxone sodium to intragastic administrate Balb/c mice, we discovered dominant microbe decreased obviously and establish light micro dysbiosis; meanwhile utilize 8g/kg/d doses, dominant microbe decreased to zero, establishing heavy micro dysbiosis.3.Compared with normal mice, the levels of TLR9 was lower and TLR4 and TLR2 were higher of mRNA in dysbacteria mice, moreover the heavy-dysbacteria mice were higher than light-dysbacteria mice.4.Compared with normal mice, the levels of NF-κBp65 was up-regulation of both mRNA and protein in dysbacteria mice, moreover the heavy-dysbacteria mice were higher than light-dysbacteria mice.Conclusion: Use different doses of ceftriaxone sodium to intragastic administrate Balb/c mice, we establish mice model for heavy and light dysbiosis of intestinal microflora; compared with normal mice, the levels of TLR9 was lower and TLR4 and TLR2 were higher of mRNA in dysbacteria mice; NF-κBp65 was up-regulation of both mRNA and protein.