| Chapter 1 The effects of B. adolescentis BF0624 and L. acidophilus LT0637 on murine acute and convalescent ulceraitve colitisObject:To establish DSS-induced acute and convalescent ulcerative colitis(UC) models and then investigate the effect of B. adolescentis BF0624 and L. acidophilus LT0637 on UC,which provides experimentMethods:50 specific pathogen free,20±2g,6-8 week-old BALB/c female mice(HuNan, certificate:SCXK2002-003), were randomly allocated in 10 groups:acute normal control group(NS-7), L. acidophilus LT0637-treated group(LT0637-7), B.adolescentis BF0624-treated group(BF0624-7),SASP-treated group(SASP-7), NS control group(NS-7); convalescent normal control group(NS-17), L. acidophilus LT0637-treated group(LT0637-17), B. adolescentis BF0624-treated group(BF0624-17), SASP-treated group(SASP-17) and NS control group(NS-17). With the exception of normal control mice,40 BABL/c mice were induced by 2.5% DSS(MW:36000-50000, MP Biomedicals Inc) water for 7 days free drinking. BF0624-7/-17 mice were given 0.2ml BF0624 via gavage once a day for 7 days(acute) or 17 days(convalescent). The mice of LT0637-7/-17, SASP-7/-17, NS-7/-17 group were respectively received instead 0.2ml LT0637/SASP/NS for 7/17 day via the same procedure, while normal controls let untreated. The animal were recorded daily both weight,fecal trait and bleeding(o-toluidine). The animals were killed on 8th day(acute) or 18th day(convalescent) morning. The whole colon(from the colo-cecal junction to the anus)was isolated for examination of length and pieces for hematoxylin and eosin stain, and histology scores were measured. According to these data, to evaluate the effects of two strains of probiotics on murine acute and convalescent ulcerative colitis.Result:1. The DSS-drinking mice were hair-withered, depressed,sluggish, stool-thinned even diarrheic,OB positive and weight-loss.After LT0637, BF0624 and SASP treatment, body weight loss,diarrhea and haemic feces was descented. LT0637 was found more effective on protecting weight loss at the acute stage than BF0624 or SASP,while BF0624 was more effective at the recovery stage.2. The DAI scores of DSS-mice increased significantly,and decreased by LT0637,BF0624 and SASP treatment.No difference in scores was found among groups at the acute stage,but the scores of LT0637-treated group were lower than that of BF0624-or SASP-treated group at the recovery stage.3,At the acute stage of DSS-mice ulcerative colitis, the thickness of colon wall reduced, feces in the enteric cavity was unformed and haemic, the length of colon reduced obviously than that of normal mice (8.10±0.52cm). In group LT0637-7, BF0624-7 and SASP-7, the decrease of colon length was prevented. At the recovery stage, the feces status improved, the length of colon increased than that of the active stage. In group LT0637-17, BF0624-17 and SASP-17, the colon length were all longer than that in group NS-17, but still shorter than that in group N-17. 4,The colonic tissue of DSS-micel under microscope showed incompleteness of mucosa, damage of recess, hemorrhage of mucosa and slight infiltration of inflammatory cells. At the active stage, BF0624 and SASP didn't show obvious beneficial effects of colonic tissue, but LT0637 showed decrease of recess damage and mucosa hemorrhage. The histology scores of each group was:8.8±1.1 of NS-7 group,7.4±1.9 of SASP-7 group,7.6±2.5 of BF0624-7 group and 6.0±1.0 of LT0637-7 group, all of these were higher than the scores at 0 of N-7 group (P<0.05), the score of LT0637-7 group was under NS group (P<0.05). At the recovery stage, the histology scores of LT0637-17 group, BF0624-17 group and SASP-17 group were higher than that of N-17 group (P<0.05), but under NS-17 group (P<0.05).Conclusion:1,The acute and convalescent UC induced by DSS showed features of active stage and recovery stage of mild to moderate UC with low mortality, which could be used in research of mild and moderate UC.2,The B. adolescentis BF0624 and L. acidophilus LT0637 can reduce the inflammation of active UC and maintain remission, the effect of the two stains are different at the two stages. L. acidophilus LT0637 had benefits to the release of active UC while B. adolescentis BF0624 had benefits to maintain remission. Chapter 2 The effects of B. adolescentis BF0624 and L. acidophilus LT0637 on proportion of Thl/Th2 in acute and convalescent UC-miceObject:To observe the proportion of Thl/Th2 in peripheral blood of UC-mice in acute stage and recovery stage and analyze the effect of B. adolescentis BF0624 and L.acidophilus LT0637 on the proportion of Th cell at two stage.Method:Experimental animal,groups, modeling and dosage regimen were same as chapter 1. The blood of mice were collected by removing their eyes in the end of expetiment, and then the mononuclear cells were isolated.The proportion of Thl/Th2 was analyze through flow cytometry,by labeling the specific antibody of Th cellular membrane with the CD4 antibody, and the cytoplastic antigen of Th1 or Th2 with IL4 antibody or IFN-γantibody.Result:1,The proportion of Thl/Th2 in normal mouse is 0.84-0.94,and it raises up in DSS-mice at both acute stage and recovery stage.2,the proportion of Th1/Th2 decreased unequally after 7 or 17 days' B. adolescentis BF0624, L. acidophilus LT0637 and SASP treatment, but that of all three groups were lower than NS group (P<0.05).And the effects of two probiotics-treatedt groups on Thl/Th2 were better than that of SASP group (P<0.05).Conclusion:The proportion of Th1/Th2 increased at the acute stage and recovery stage of DSS-mice. Both B. adolescentis BF0624 and L. acidophilus LT0637 had more effective than SASP on decreasing the proportion of Th1/Th2 at two stages.Chapter 3 The immunologic mechanism of B. adolescentis BF0624 and L. acidophilus LT0637 in treating acute and convalescent UC-miceObject:To analyse the immunologic regulation of the B. adolescence BF0624 and L. acidophilus LT0637 in treatment of acute convalescent ulceratives colitis mice.Method:Experimental animals,grouping,modeling and dosage regimen were same as chapter 1.Sent the mice to death when the experiments at the acute and recovery stage done,then kept the colon of every group. The level of TLR2,TLR4,PPARy,NF-κB and T-bet in the colon of experimental mice were determined by Western blotting and semiquantitative RT-PCR methods.Result:1,The level of TLR2,TLR4,NF-κB and T-bet on colonic mucosa increased obviously (P<0.05) when the level of PPARy decreased obviously(P<0.05) both at the acute stage and recovery stage.2,The level of TLR2,TLR4,NF-κB and T-bet decreased while the level of PPARy increased(P<0.05) after 7/17 days'B. adolescence BF0624, L. acidophilus LT0637 or SASP treatment.Conclusion:1,B. adolescence BF0624 and L. acidophilus LT0637 could inhibit the expression of TLR2,TLR4 at the acute stage and recovery stage, then reduced the expression of NF-κB and T-bet.2,B. adolescence BF0624 and L. acidophilus LT0637 could activate the expression of PPARy to alleviate intestinal tract inflammtion of UC and maintain remission.
|
PDF Full Text Request