| Objectives: By observing the effect of Lan Yin Feng Yang Hua Zhuo Jie Du Decoction for Ulcerative Colitis(Ulcerative Colitis, UC) model rats on general conditions, disease activity index(DAI) score and pathological histology and Intestinal Trefoil Factor(ITF),Epidermal Growth Factor(EGF), Extracellular Regulated Protein Kinase(ERK),the key proten of Mitogen-Activated Protein Kinase(MAPK),in the colon tissue,the purpose of the study was to explore the mechanism for the treatment of UC rats,and provided theoretical and experimental basis in the clinical application.Methods:60 healthy male rats(weighing200g-250g)were randomly divided into 6 groups after adaptive feeding for 7days:the normal group(NG), the model group(MG), the high-dose, the middle-dose and the low-dose Lan Yin Feng Yang Hua Zhuo Jie Du Decoction group( HDG,MDG and LDG), the Srlfasalazine group(SASP).Each group included 10 rats. Besides NG, the other five groups adopted 2, 4, 6-trinitrobenzene sulfonic acid/ethanol(TNBS/ethanol) to prepare UC rats model. 3 days after the building, NG rats in the normal feeding, saline lavage to MG rats, SASP rats using Srlfasalazine Mixed suspension lavage,the Chinese medicine treatment groups used the corresponding drug lavage.After 14 days of continuous treatment, observed the state of mind,drinking and diet, activity, weight, excrement characters, and so on and so forth of rats in each group every day.After the treatment, each group of rats were giwen fasting and water for 24 hours,and recorded their weight.Observed the general view of the colonic mucosa and histopathological changes after all rats were sacrificed. Measured the expression of ITF,EGF,and MAPK/ERK by RT-PCR method.Results:1 The general conditions of ratsRats in NG had good mental state, glossy fur, normal eating,drinking and activity.their faeces was granular without mucus,pus and blood.Body weight increased gradually. The rest of the 5 groups of rats gradually appeared listlessness, disheveled and matt fur,burnout and crowed state,reduced activity and diet, loose stool or even stool with pus and blood and filthy pad after modeling. The symptoms of NG rats worsened gradually with the increase of experimental days. Even perianal appeared visible to the naked eye bloody stool and body weight of rats alleviated in different degrees. Mental situation of rats in the other four treatment groups,fur colour and lustre, activity, weight and so on were improved after drug intervention. Rats in HDG improved the most significantly and general situation was close to NG. There was no evident difference between SASP and MDG in general condition, General condition in SASP and MDG were lower than HDG and higher than LDG. General condition in LDG was slightly higher than MG.2 DAI scoreIn terms of disease activity index score, there was statistically significant difference(P<0.05) between the treatment group HDG(0.37±0.35),MDG(0.81±0.29),LDG(1.56±0.41),SASP(0.78±0.29)and MG(2.46±0.43). There was statistically significant difference(P<0.05) between HDG and MDG,SASP,LDG. There was no statistically significant difference(P>0.05) between SASP and MDG. There was statistically significant difference(P<0.05) between SASP and LDG.3 Macroscopic histopathological observation of colonic mucosaThe naked eye observation of MG in colonic mucosa was the worst case.There was much ulcer.HDG was approach to NG.MDG and SASP were slightly better, and MDG was better than SASP. LDG was inferior to the other 3 treatment groups with interspersed ulcer, but better than MG.Histopathological observation: Mucosa in NG was smooth and lustrous. Mucosal glands in MG,which were infiltrated by a large number of inflammatory cells, along with mucosal hyperemia and edema, were damaged.Mucosal destruction situation in LDG was inferior to MG. There were basically complete mucomembranous structure in LDG,which was closed to HDG.The situation of MDG was inferior to HDG. The outcome of SASP was between HDG and MDG,but better than that of LDG.4 Comparison of colon tissue ITF expressionThe relative expression value of ITFm RNA was calculated from the gray seale ratio of ITF and GAPDH. ITFm RNA in MG(0.24±0.02) below NG(0.27±0.01), the difference was statistically significant(P<0.05); The treatment group were higher than that of MG, the difference was statistically significant(P<0.05); HDG(0.58± 0.03) higher than that of SASP(0.55±0.02), MDG(0.50±0.03) and LDG(0.43±0.03), the difference was statistically significant(P<0.05); MDG higher than that of LDG, the difference was statistically significant(P<0.05); SASP was higher than MDG, LDG with statistically significant difference(P<0.05).5 Comparison of colon tissue EGF expressionThe relative expression value of EGFm RNA was calculated from the gray seale ratio of EGF and GAPDH. EGFm RNA in MG(0.26±0.04) compared with that in NG(0.24±0.03), the difference was not statistically significant(P>0.05); There was no statistically significant difference between HDG(0.42±0.42) and SASP(0.41±0.02)(P>0.05).MDG(0.36±0.02),LDG(0.33±0.03) were less than HDG with statistically significant difference(P<0.05); LDG was lower than MDG wit statistically significant difference(P<0.05); MDG and LDG were lower than SASP with statistically significant difference(P<0.05).6 Comparison of colon tissue MAPK/ERK expressionThe relative expression value of MAPK/ERKm RNA was calculated from the gray seale ratio of MAPK/ERK and GAPDH. MAPK/ERKm RNA of MG(0.27±0.01) was higher than that of NG(0.24±0.01) with statistically significant difference(P<0.05); The treatment group compared with MG, the difference was statistically significant(P<0.05); HDG(0.51±0.02) was higher than that of SASP(0.49±0.01), MDG(0.48±0.11), LDG(0.41±0.01) with statistically significant difference(P<0.05); SASP compared with MDG, the difference was not statistically significant(P>0.05); LDG was lower than MDG with statistically significant difference(P<0.05).Conclusion:1 Lan Yin Feng Yang Hua Zhuo Jie Du Decoction can improve the general condition, alleviate the pathological damage and reduce the DAI score of UC rats effectively. And there is a dose-effect relationship between its curative effect and dosage: The effect of high doses group is much better than the medium doses group and the low doses group of treatment.2 The mechanism of action of Lan Yin Feng Yang Hua Zhuo Jie Du Decoction to treat UC is probably likely to improve expression of ITF,EGF, and further activate the MAPK/ERK in colon tissue, inhibit the infiltration of inflammatory cell effectively, repair the damage of intestinal mucosa, so as to achieve the purpose of treatment of UC. |
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