Effects Of Qingre Huashi Quyu Recipe On Oxidative Stress And Apoptosis Of Colonic Mucosa In UC Mice

Ulcerative colitis(UC)is a disease characterized by recurrent abdominal pain,diarrhea,mucus,pus and bloody stool.The disease seriously affecting the quality of life of patients at a high incidence,for a long course and difficult to heal.TCM treatment of UC has significant advantages.Qingre Huashi Quyu Decoction has shown good therapeutic effect in clinical practice by playing the role of clearing heat and removing dampness,promoting blood circulation and removing blood stasis,but its mechanism is still unclear.Therefore,this project intends to explore the curative effect and mechanism of Qingre Huashi Quyu Decoction on UC model mice through animal experimental research,so as to provide a new idea for the treatment of UC.Experiment 1:Effect of Qingre Huashi Quyu Decoction on ulcerative colitis in miceObjective:To observe the therapeutic effects of Qingre Huashi Quyu Decoction on ulcerative colitis(UC)using the mice ulcerative colitis model which was induced by dextran sodium sulfate(DSS).Methods:Thirty two C57BL/6 mice were divided into normal group(n=8)and operation group(n=24)according to the ratio of 1:3.The operation mice were divided into model group,mesalazine group and traditional Chinese medicine group according to the ratio of 1:1:1,with 8 mice in each group.Except for the normal group,the model mice were free to drink drinking water containing 3%DSS for 6 days to prepare UC mice model.After that,all mice were changed to drink distilled water.The mesalazine group was given mesalazine suspension 100mg/kg/d,the traditional Chinese medicine group was given Qingre Huashi Quyu Decoction 15.925g/kg/d,the normal group and the model group were given equal volume of 0.9%sodium chloride solution,once a day,for 7 consecutive days.The body weight,stool texture and fecal occult blood of each mouse were observed and recorded daily,and the disease activity index(DAI)of each mouse was calculated.After the treatment,the mice were killed,the colon segment from anus to end of cecum was cut,and the colon length was measured.Then the morphological changes of colon tissue were observed by HE staining,and the histopathological score was made.Results:1.DAI score:after DSS induced UC mice model,there were different degrees of weight loss,stool texture changes and fecal occult blood.Compared with the normal group,DAI score of the model group was significantly increased(P<0.05).Compared with the model group,the DAI scores of mesalazine group and Chinese medicine group were significantly lower on the 8th and 9th day(P<0.01).2.Colon length:compared with the normal group,the colon length of the model group was significantly shorter(P<0.05);Compared with the model group,the colon length of mesalazine group and traditional Chinese medicine group was significantly longer(P<0.05).3.HE staining results and pathological score of colon tissue:histological analysis showed that the mice in the model group showed obvious colonic epithelial damage,including mucosal erosion,crypt loss and extensive lymphocyte infiltration;Mesalazine and traditional Chinese medicine treatment significantly improved these pathological changes.Compared with the normal group,the pathological score of the model group was significantly higher(P<0.05);Compared with the model group,the pathological scores of mesalazine group and traditional Chinese medicine group were significantly lower(P<0.05).Conclusion:Qingre Huashi Quyu Decoction has a certain protective effect on DSS induced UC mice,and the effect is similar to that of mesalazine.Experiment 2:Effect of Qingre Huashi Quyu Decoction on oxidative stress and apoptosis of colonic mucosa in mice with ulcerative colitisObjective:To explore the possible mechanism of qingrehuashi Quyu Decoction in the treatment of UC by detecting the oxidative stress and apoptosis in colon tissue of UC mice model induced by dextran sulfate sodium(DSS).Methods:Thirty two C57BL/6 mice were divided into normal group(n=8)and operation group(n=24)according to the ratio of 1:3.The operation mice were divided into model group,mesalazine group and traditional Chinese medicine group according to the ratio of 1:1:1,with 8 mice in each group.Except for the normal group,the model mice were free to drink drinking water containing 3%DSS for 6 days to prepare UC mice model.After that,all mice were changed to drink distilled water.The mesalazine group was given mesalazine suspension 100mg/kg/d,the traditional Chinese medicine group was given Qingre Huashi Quyu Decoction 15.925g/kg/d,the normal group and the model group were given equal volume of 0.9%sodium chloride solution,once a day,for 7 consecutive days.After the treatment,the mice were killed,and the colon from the anus to the end of cecum was cut and divided into four equal parts,according to the requirements of different experiments were treated differently.For the observation of oxidative stress in colonic tissue,the content of ROS in colonic tissue of mice was detected by DHE fluorescence staining.The content of MDA and the activity of SOD in colonic tissue of mice were detected by malondialdehyde(MDA)test kit and superoxide dismutase(SOD)test kit.For the observation of apoptosis in colon tissue,TUNEL fluorescence staining was used to detect the level of apoptosis in colon tissue.Results:1.Oxidative stress:Compared with normal group,ROS and MDA in colon tissue of model group were significantly increased(P<0.05),SOD activity was significantly decreased(P<0.05);Compared with the model group,the expression of ROS and MDA in mesalazine group and traditional Chinese medicine group were significantly decreased(P<0.05),and the activity of SOD was significantly increased(P<0.05).2.Apoptosis:Compared with the normal group,the level of apoptosis in the model group was significantly increased(P<0.05);Compared with the model group,the level of apoptosis in mesalazine group and traditional Chinese medicine group was significantly lower(P<0.05).Conclusion:There are oxidative stress and excessive apoptosis of intestinal epithelial cells in the colonic mucosa of UC mice.The protective effects of Qingre Huashi Quyu Decoction on UC may be related to regulating oxidative stress and improving apoptosis of colonic epithelial cells.