Effects Of Tetramethylpyrazine On Exprimentalul Cerative Colitis In NF-kB/COX-2Signal Pathway

Objective：The purpose of this study was to detect the change of NF-kB/COX-2signaling pathway in experimental colitis before and aftertetramethylpyrazine treatment, to study the effects of Ligustrazine onulcerative colitis and explore its mechanism.Methods:1. To establish the animal model of ulcerative colitisThe mice model of ulcerative colitis was induced by5%dextransulfate sodium (DSS), all mouse with oral feeding.The mouse wererandomly divided into four groups:normal control group (group N), modelgroup (group M),saline treatment control group (group Y), Ligustrazinetreatment group (group Z). To evaluate the UC model by the generalobservation, disease activity index (DAI) and the scores of histologicalchanges.2.The relationship between the change of NF-kB/COX-2signalpathway and ulcerative colitisTo evaluate the changes of NF-kBp65, COX-2expression during UC occurred by Reverse Transcription-Polymerase Chain Reaction（RT-PCR)and immunohistochemical methods。3.The intervention effect of Ligustrazine for the NF-kB/COX-2signaling pathway in ulcerative colitisThe effects of ligustrazine in treating ulcerative colitis was comparedto saline treatment group and UC model group, to observe the expressionchanges of NF-kBp65, COX-2in treatment process, to investigate theeffects and mechanism of ligustrazine in treating ulcerative colitis.Results:1. general situation：During the trial period, normal control groupdrinking water in normal and fur shiny, vibrant, responsive, no stoolchanges, and no fecal occult blood test was positive. In the modelgroup:after using5%DSS solution instead of drinking water twodays,several mouse began to appear positive fecal occult blood,but noobvious changes of stool.The mouse became to appear stool thinning,positive fecal occult blood, even visible brown stool, several mouse couldbe found a little sticky anal, dietary reduction, easy frightened, like to staytogether in the3-4day;5-7day appeared significantly dilute stool, evendilute pure blood stool could be found by the naked eye,diet decreases, furmessy little luster, spirit is dispirited, apparent weight loss. Anatomicalobservation: colon of the control mice was pink, bowel wall wassmooth;colon of model mice was dark red, intestinal mucosa hyperemia edema, or even visible intestinal cavity has a large number of bloodysecretion.2.HE staining: model group mice colon mucosa showed obviousinflammatory response, mainly off the surface epithelium, extensiveinflammatory cell invasion, glandular atrophy or disappearance and othertypical inflammatory response. The histological differences between themodel group and the control group had statistically significant, thehistological change of M group consistented with ulcerative colitisfeatures.Tip:the modelis succeed. After treatment of7days, TMP treatmentgroup compared with model group and treated group,the visible mucosalcongestion and edema reduced, there are still some neutrophil, lymphocyteinfiltration in endoscopic mucosal and mucosal shallow,but the histologicalscores in TMP treatment group was lower than the saline treatment groupand model group(P <0.05).3. The results of RT-PCR and immunohistochemistry: the mRNA ofNF-kBp65and COX-2expression were significantly increased in M groupwhich compared with N group,,(P <0.05), after the applicationtetramethylpyrazine treatment of UC, the inflammation evaluation indexwere significantly decreased than those in M group, the mRNA ofNF-kBp65and COX-2expression were significantly reduced (P<0.05).After saline treatment, inflammation evaluation index has improvedin Y group than those in M group, but the results are not statistically significant. The Immunohistochemistry displayed NF-kBp65of COX-2expression were significantly increased in the M group,aftertetramethylpyrazine treatment, the inflammation evaluation index and theexpression of NF-kBp65and COX-2was significantly reduced,the resultsin Z group compared with the M group group had statistically significant, Ygroup compared with Z group, the inflammation evaluation index and theexpression of NF-kBp65, COX-2was significantly increased, the resultshad no statistically significant (P <0.05).Conclusion:The expression of NF-kBp65and COX-2had increased in the UCmodel mice, the overexpression of them may cause intestinal inflammationexacerbation, ulcerative colitis deteriorated. Indicates that the NF-kB/COX-2signaling pathways involved in the development of ulcerativecolitis.Tetramethylpyrazine can change the expression of NF-kBp65andCOX-2, reverse blocking the deterioration of ulcerative colitis. Promptedthat the Ligustrazine in treating ulcerative colitis through the signalingpathway of NF-kB/COX-2, but it may not be completely dependent onthis signal pathway.