| Objective:To establish a rat model of Ulcerative Colitis(UC)caused by spleen de ficiency and dampness and liver stagnation syndrome,and to study the intervention eff ect of Jiuxiening Granule on UC model rats with dampness and liver stagnation syndr ome caused by spleen deficiency and reveal the possible mechanism of its treatment o f Ulcerative Colitis.Methods:1.SPF grade SD rats aged 7 weeks were divided into Dextran sulfate sodium salt(DSS)group,TCM syndrome group and combined disease group.The rats were given quantitative gavage of 10%DSS solution with emotional,dietary and environmental intervention,and quantitative gavage of 10%DSS solution + emotional,dietary and environmental intervention,respectively.General conditions(body weight,mental state,movement behavior,hair brightness,fecal traits,etc.)of rats in each group were observed during modeling,and Disease Activity Index(DAI)score was performed.After building with enzyme-linked immunosorbent assay(Elisa)to detect each group rats serum interleukin 2(IL-2)and tumor necrosis factor alpha(TNF-α)content,macroscopic observation of Colon tissue morphological changes,and colonic mucosa damage Index(Colon Mucosal D-amage Index,CMDI)score,the microscopic observation rat Colon tissue pathological evaluation model,to determine the best method of building.2.Duplicate the model and randomly divide the SD rats into model control group,positive control group,Jiuxiening granules group,and normal control group.After modeling,drug intervention was performed.Normal control group and model control group were given normal saline intragastrically.The positive control group was gavage with equivalent salazosulfopyridine solution.According to the relevant pharmacodynamics experimental study in the early stage,the optimal pharmacodynamic dose group was selected,and the corresponding dose of Jiu Xieting Granules water decoction was given daily by gavage;14days in a row.Groups of rats at the end of the dosing with immunohistochemical staining method,protein imprinting method to detect each group rats colon tissue inner adenosine phosphate/protein kinase A(PKA/c AMP)signaling pathway,water channel protein 4(AQP4)and water channel protein 8(AQP8)distribution,expression,and relative content,using reverse transcription polymerase chain reaction method(RT-PCR)detection of each group rats colon tissue AQP4 and AQP8 m RNA expression levels.Results:1.After modeling,the body mass of the disease-syndrome combination group decreased significantly,including listlessness,huddling,drowsiness,lazy movement,weak cry during capture,sparse hair and easy to fall off,sparse stools,occasional bloody stools,perianal filth,and fecal occulting blood(+).The DAI and CMDI scores were significantly higher than those of the normal control group(P < 0.01).Serum IL-2 and TNF-α content,compared with normal control group with significant difference(P < 0.01),then to observe the pathological colon mucosa shows marked hyperemia and edema,visible to the naked eye sores,mucosa canker,glandular structures destroyed,goblet cells disappeared,a large number of inflammatory cells infiltration,interstitial edema,congestion and inflammatory cells infiltration.Building 14 days after the completion of the group of rats colon tissue combined disease,macroscopic observation colonic mucosal surface is still congestion,edema,scattered in the ulcer point visible tip samples,the mucosa is still have defects,arrange irregular glands,part of the structure damage,goblet cells decreased,a large number of inflammatory cells infiltration,interstitial edema,congestion,a small amount of inflammatory cells infiltration.2.After the intervention of Jiuxiening granules,the body weight of SD rats was significantly increased,the spirit was good,the diet amount was improved,the feces were oval,soft,occulted blood(-),and the hair was hae.Compared with the model group,the CMDI score of Jiuxiening Granules group was significantly decreased(P < 0.01).Some colonic mucosal tissues were in a state of repair,the glands were neatly arranged,the goblet cells increased,and the interstitial edema and congestion were significantly reduced.The contents of IL-2 and TNF-α in serum of rats were significantly different from those of model group(P < 0.01).The average optical density of AQP4,AQP8,c AMP and PKA in the colon of SD rats was significantly lower than that in the normal control group by immunohistochemical detection(P < 0.01);Compared with the model control group,the positive control group and Jiuxiening granules group were significantly increased(P < 0.01).Compared with the normal control group,the protein expressions of AQP4,AQP8,c AMP and PKA in the colon of SD rats were significantly decreased in the model control group(P <0.01).Compared with the model control group,the positive control group and Jiuxiening granules group were significantly increased(P < 0.01).RT-PCR showed that the m RNA expressions of AQP4 and AQP8 in colon tissue of SD rats were significantly decreased compared with the normal control group(P < 0.01).Compared with the model control group,the positive control group and Jiuxiening granules group were significantly increased(P <0.01).Conclusion:1.Quantitative gavage of 10%DSS solution combined with emotion,diet and environment intervention was used to simulate the ulcerative colitis rat model of spleen deficiency,dampness and liver stagnation syndrome combined with disease and syndrome.The success rate was high,and the model was stable with both pathological characteristics of UC and characteristics of TCM syndrome type.2.Jiuxieting Granule is effective in the treatment of ulcerative colitis caused by spleen deficiency and dampness and liver depression syndrome.It can repair damaged colonic mucosal tissue and restore its physiological function and its mechanism may be related to the regulation of aquaporin 4 and 8 expression by regulating c AMP/PKA signaling pathway in colon tissue. |
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