The Influence Of Oxymatrine On TLR9Pathway And The Improvement Of The Lipid Peroxidation In Ulcerative Colitis Rats Models

Objective:In this experiment we observed the expression of TLR9/Myd88/NF-κB mRNAin rat model of Ulcerative Colitis(UC), and we investigated the influence ofOxymatrine(OMT) and Mesalazine on TLR-9/Myd88/NF-κB-dependentinflammatory responses in UC,and we also observed the protective effects ofoxymatrine and Mesalazine during the lipid peroxidation in UC by detecting SOD andMDA.This study will explore the mechanisms of anti-inflammatory and resisting lipidperoxide effects on OMT in UC.Methods:We used75SPF grade of adult SD rats,male,and then randomly divided into5groups randomly: control group, OMT group, Mesalazine group, OMT+Mesalazinecombined group, and model group. After fasting24hours, the rats were anesthetized,then with the silicone tube inserted in the anal about8cm,clysis with injectingtrinitrobenzene sulfonic acid (TNBS,100mg/kg) and ethanol solution.The followingday began all the rats were fed on oral gavage OMT or Mesalazine, the model groupand the control group were fed on with the same dose of distilled water. One weeklater,three rats in each group were randomly selected and executed for observingcolonic histological changes.On day15, the remaining rats were executed afterfasting24h to detect the expression of TLR9/Myd88/NF-κB mRNA in colon tissueby RT-PCR,and detect the activities of SOD and the content of MDA.Results:1. General shape and pathology test results: the model group had colonic mucosacongestion, edema, erosion, necrosis, bowel wall thickening, ulceration. The results oflight microscope observation shows a large number of neutrophils, plasma cells and other inflammatory cell infiltrated. Colonic epithelial cell line structures aredisorganized,with glands damaged.The treated groups' had a relieved inflammationexpression with colonic mucosa congestion, edema, the ulcers are small andsuperficial,under the light microscope the inflammatory cell infiltration are improvedthan the model group,also we observed a proliferation of colonic glands. Histologicalexamination confirmed the diagnostic results. The control group rat colon tissueneatly arranges on each floor,with few inflammatory cells.2. RT-PCR results: The expression of TLR9, Myd88, and NF-κB mRNA in UCmodel rat colon tissue are higher than in treated groups and normal control group, thedifference was statistically significant (p<0.01).In the treated groups,the OMT+Mesalazine combined group shows a lower expression of TLR9, Myd88and NF-κBmRNA than the OMT group and the Mesalazine group, the difference was statisticallysignificant (p<0.05),but between the OMT group and the Mesalazine group,thedifference has no statistically significant (p>0.05).3.SOD,MDA results:In UC model group,the activity of SOD shows a lower leverthan all the other groups,while the content of MDA shows a higher lever than theother groups, the difference was statistically significant (p<0.05). In the treatedgroups,the combined group shows a higher SOD activity,a lower MDA content thanthe other2groups, the difference was statistically significant (p<0.05), between theOMT group and the Mesalazine group,the difference has no statistically significant (p>0.05).Conclusion:1.The OMT and Mesalazine combined use have a better effect on blocking theTLR9/Myd88/NF-κB pathway to alleviate the inflammatory reaction in UC thanOMT or Mesalazine use alone.2. The OMT can increase the activity of SOD and decrease the content of MDAin UC model rat colon tissues,shows a better resisting lipid peroxide effect by OMTand Mesalazine used together.