Experimental Study Of Qingdai On The Treatment Of DSS-induced Ulcerative Colitis In Mice

Purpose:Through studying the expression of Inflammatory Factors TNF-?,IL-6,TGF-?1,IL-10 and apoptosis factors Bcl-2,Bax and Caspase-3 protein and mRNA expression in mice with ulcerative colitis induced by dextran sodium sulfate(DSS),to explore the therapeutic effect of Indigo Naturalis on DSS-induced ulcerative colitis in mice.Material and method:Sixty healthy male C57BL/6 mice(weight 20-24g)were randomly divided into 20 control groups and 40 model mice.Model mice were free to drink 5%DSS solution for 7 days to induce UC model.The next day after the successful model building,the model mice were randomly divided into a model group and an Indigo Naturalis group.The control group and the model group were given 0.5%sodium hydroxymethyl cellulose by gavage every day,and the Indigo Naturalis suspension made of 0.5%sodium hydroxymethylcellulose was given by gavage.The gavage volume was both 10mL/kg,continuous 10 days.After the last administration,the mice were fasted and water for 24 hours,the eyeballs were removed and blood was taken,and all the mice were sacrificed.Observe the general state of the mouse;record the disease activity index of the mouse;observe the changes in the morphology of the mouse colon under a light microscope;observe the mucosal tissue of the mouse colon by hematoxylin-eosin staining Changes in the study;enzyme-linked immunosorbent assay method to detect the expression of TNF-?,IL-6,TGF-?1 and IL-10;TdT-mediated dUTP Nick-End Labeling method to observe cell apoptosis in mouse colon tissue Changes in the number of deaths;Western blot detection of Bcl-2,Bax and Caspase-3 protein expression;Real-time polymerase chain reaction detection of Bcl-2,Bax and Caspase-3 mRNA expression.Results:1.General conditions of mice and DAI score:The mice in the control group have white coat color with brilliance,quick response,normal exercise and diet,basically no change in urine output,and no blood in the stool.The mice in the model group have dry and dull coat,mental atrophy,reduced activity,indifference to food,diarrhea,mucus pus and blood in the stool.The hair color,exercise and diet of the mice in the Qingdai group were better than those in the model group.The DAI score results showed that compared with the control group,the DAI score of the model group was significantly higher than that of the control group,and the difference was statistically significant(P<0.05);compared with the model group,the DAI score of the Indigo Naturalis group was significantly lower,and the difference was statistically significant Scientific significance(P<0.05);There was no statistically significant difference between the Qingdai group and the control group(P>0.05).There was no significant difference between the Qingdai group and the control group(P>0.05).2.Histopathological changes:The mucosal surface of the mice in the control group is smooth,without hyperemia,edema,and the glands are arranged regularly.The mouse colonic mucosal epithelial cells in the model group had degeneration,necrosis,hyperemia,edema,ulcers,and inflammatory cell infiltration.In the Indigo Naturalis group,mild degeneration and necrosis of epithelial cells and a small amount of inflammatory cells were seen.The histological score results showed that compared with the control group,the histological score of the model group was significantly higher than that of the control group,and the difference was statistically significant(P<0.05);compared with the model group,the histological score of the Indigo Naturalis group was significantly lower.The difference was statistically significant(P<0.05);there was no statistically significant difference between the Qingdai group and the control group(P>0.05).There was no significant difference between the Qingdai group and the control group(P>0.05).3.Index detection:The expression levels of TNF-?,IL-6,and TGF-?1 in the serum and colon of the model group were significantly higher than those in the control group,and the IL-10 expression levels were lower than those in the control group.The difference was statistically significant(P<0.05).The expression of Bcl-2 gene and protein in the model group was lower than that of the control group,while the protein and mRNA expression of Bax and Caspase-3 was significantly higher than that of the control group,and the difference was statistically significant(P<0.05).After treatment with Indigo Naturalis,compared with the model group,the expression of TNF-?,IL-6 and TGF-?1 in the serum of mice was significantly reduced,and the expression of IL-10 was significantly increased,and the difference was statistically significant(P<0.05).The expression of Bcl-2 protein and mRNA was significantly increased,while the expression of Bax and Caspase-3 protein and mRNA was significantly decreased,and the difference was statistically significant(P<0.05);compared with the control group,the Qingdai group had no statistically significant difference(P>0.05).There was no significant difference between the Qingdai group and the control group(P>0.05).Conclusion:1.Indigo Naturalis can relieve the symptoms of mice and relieve the damage of the intestinal mucosa.2.Indigo Naturalis can significantly affect the expression of inflammatory mediators TNF-?,IL-6,TGF-?1,IL-10 and apoptosis factors Bcl-2,Bax,Caspase-3 protein and mRNA in mice with ulcerative colitis induced by DSS.Regulates the body's immune function,inhibits excessive cell death,and has a therapeutic effect on UC.