| Objective:In this study,TNBS / ethanol mixture enema copy Chronic ulcerative colitis model SD rats after administration Shishenwan suspensions treatment efficacy Shishenwan Ulcerative Colitis was evaluated.Further verification while Shishenwan treatment of experimental rats,the intended regulation of Rho / ROCK signaling pathway angle,explore the mechanism Shishenwan Ulcerative Colitis colonic mucosa barrier.Methods:1.Animal grouping and model replication and drug delivery treatment: SD rats were randomly divided into four groups,Normal,model group(TNBS),Sishenwan group(TNBS+SSW),mesalazine group(TNBS+ASA),8 per group and each group of rats was numbered at the root of the rat tail.The normal group was given 0.8 ml saline enema.In addition to the normal group,3 mg/kg TNBS/50% ethanol suspension 0.8ml enema was administered.And the above model making method is repeated twice a week at a fixed time.From the second day after the model copy ends,the normal group and the model group were given 2 ml of normal saline,and the Sishenwan group was given.5g/kg Sishenwan suspension was intragastrically administered,and mesalazine enteric-coated tablets were administered with 150 mg/kg mesalazine enteric-coated tablets.Execution after one week of administration,the colon was sacrificed.2.Evaluate the efficacy Shishenwan treatment of ulcerative colitis in rats: weighed daily,and observe its food and water,activity,mental state and symptoms.The colon was isolated,its weight and length were measured,its intestinal weight index was calculated,and the colonic mucosal injury index score was visually observed Histopathological sections were stained with H&E staining,and histopathological changes were observed under the microscope.3.Rho/ROCK signaling,colonic mucosal barrier connexin,and cellular energy metabolism and cell differentiation-related protein analysis: Western blot analysis of Rho/ROCK signals(eg,p Rho A,ROCK 1,p Rac),and colonic mucosa Barrier-associated connexins((Claudin-5,Jam-1),adhesion-linked(VE-cadherin),energy metabolism key protein p AMPK ?,etc.From the regulation of Rho/ROCK signal,explore the mechanism of Sishenwan to maintain the integrity of colonic mucosal barrier in rats with recurrent ulcerative colitis.4.PTEN/PI3K/Akt pathway analysis: Western blot assay was used to detect and analyze the PTEN/PI3K/Akt pathway signal.To explore whether drugs can treat TNBS-induced experimental UC in multiple directions.5.Data analysis: Statistical analysis was used to import the data into SPSS 22.0software for T-test.The data was expressed in MeanąSD.P<0.05 was a significant difference in statistical results.P<0.01 was considered as statistically significant difference.Results:1.Rat ulcerative colitis model and post-treatment performance: Compared with the Normal,the model group decreased the food intake,the activity amount also decreased,the spirit was wilting,the constitution was obviously reduced,the hair dark yellow easily fell off and the unevenness was dull.Stools are rare,and there are cases of blood in the stool.The skin around the anus is sticky and sticky.Compared with the model group,the treatment group tends to improve after administration,the food intake increases,the activity is active,and the constitution increases.In addition,the stool is somewhat different.improved blood in the stool cure,showing the therapeutic effect of Shishenwan.2.Colonic tissue of rats: Compared with the Normal,the colon length of the model group group was significantly decreased(P<0.01),and the weight of the colon was significantly increased(P<0.05).The large amount of edema and congestion in the colon cavity was visible to the naked eye.Compared with the model group group,the length of colon in the SSW AND ASA group increased significantly(P<0.01),the weight of colon decreased significantly(P<0.05),and the expression of IL-1? and TNF-? in colonic mucosa also decreased significantly(P <0.01 or P<0.05).Tip colon Shishenwan have a positive effect on the improvement of colon edema,congestion,erosion,ulceration and inflammation and other conditions.3.Rho/ROCK signal and colonic mucosal barrier-associated protein: Western blotting was used to detect colon protein.Compared with the Normal,the expression of p-Rhoa and ROCK 1 in the model group was significantly increased,while the expression of p-Rac was significantly decreased,and the expression of intestinal mucosal barrier-associated proteins Claudin 5,Jam 1,CX 43 and VE-cad was also significantly increased(P<0.01);Compared with the model group,the Sishenwan group p Rho A and ROCK 1 both decreased significantly,while p Rac increased significantly.The expression of intestinal mucosal barrier-associated proteins Claudin5,Jam 1,CX 43 and VE-cad was also significantly increased(P < 0.01).This result suggests that Sishenwan can regulate the expression of colonic mucosa-associated proteins Claudin 5,Jam 1,CX 43 and VE-cad and Rho/ROCK signals in colitis rats.4.Key proteins for energy metabolism : Western blotting detects total bacterial proteins.Compared with the normal group,the expression of p-AMPK ? was significantly decreased(P<0.01);Compared with the model group,the p AMPK ?increased significantly(P<0.01).This result suggests that Sishenwan can regulate the expression of and p-AMPK ? protein in colonic mucosa of rats with colitis.5.PTEN/PI3K/Akt signaling pathway: Western blotting detects colon protein.Compared with the Normal,PI3 K and Akt were significantly increased in the model group,while PTEN was significantly increased(P<0.01);Compared with the TNBS group,PI3 K and Akt in Sishenwan group decreased significantly,while PTEN increased significantly(P<0.01).This result suggests that Sishenwan can regulate the PI3K/Akt/PTEN signalings pathway in colonic protein of colitis rats.Conclusion:1.Sishenwan is effective in the treatment of experimental ulcerative colitis.2.Sishenwanl can effectively treat experimental ulcerative colitis by inhibiting PI3K/Akt signaling pathway,and finally regulating Rho/ROCK signal to maintain the integrity of colonic mucosal barrier to treat experimental ulcerative colitis. |
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