| Objectives: Though the observation of the influence of Xiang Lian Hua Zhuo Jie Du Decoction on the general conditions, disease activity index score,pathology change and the expression of NF-κB、COX-2 mRNA of Ulcerative Colitis model rats,try to explore the mechanism of action of the Decoction in the treatment of UC in molecular level, Aims to provide experimental foundation and theoretical basis for the clinical application of the Decoction.Methods: 60 healthy male wistar rats(weighing 200g-250g) were fed for 7days for adaptive,then divided into 6 groups randomly: the normal group(NG), the model group(MG), the high-dose(HDG), the middle-dose(MDG) and the low-dose(LDG) of Xiang Lian Hua Zhuo Jie Du Decoction group, the Srlfasalazine group(SASP), each group included ten rats. Besides NG, the other groups were adopted 2, 4, 6-trinitrobenzene sulfonic acid/ethanol(TNBS/ethanol) to prepare the UC rat models. After the success of the building, each Chinese medicine treatment group used the corresponding drug for gavage. SASP rats used the mixed suspension of Srlfasalazine for gavage, MG rats used saline for gavage, NG rats were given by the normal feeding. Observed the condition of mental state, drinking and diet, excrement characters, weight, and so on. Record and calculate the DAI score of rats in each group. After 2 weeks of continuous gavage,killed the rats,and observed the general view and histopathological changes of the colonic mucosa. Measured the expression of NF-κB、COX-2 mRNA through the method of RT-PCR(Reverse transcriotion- polymerase chain reaction,RT-PCR).Results:1 The general conditions of ratsRats in NG had normal granular stool, responsive fast, normal eating and drinking, fur luster, and weight increased gradually.Compared with NG,the other groups of rats gradually appeared loose stool, or even stool with pus and blood, filthy pad, depression, burnout and crowed state, disheveled and matt fur, activity and diet reduced obviously, slim down.The general conditions of MG rats were worst.When experiment ended,the rats in NG were in good condition. The rats in MG restored the worst. The general conditions of the other 4 groups of rats were improved in different degrees,such as stool, mental condition, fur luster, eating and drinking and weight. The general conditions of HDG and SASP rats restored the most obvious. These of MDG were worse than HDG,but better than LDG. And LDG rats were slightly better than MG.2 DAI scoreThe DAI score of NG was 0.There was statistically significant difference(P<0.05) between the treatment group HDG(0.37±0.35),MDG(0.90±0.32),LDG(1.50±0.62),SASP(0.63±0.35) and MG(2.50±0.47). There was statistically significant difference(P<0.05) between HDG and MDG,LDG. There was no statistically significant difference(P>0.05) between SASP and HDG. 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. There was statistically significant difference(P<0.05) between MASP and LDG.3 Observation of colonic mucosaThe naked eye observation of NG in colonic mucosa did not see hyperemia and edema, flexible smooth.Fold are clear and complete. Microscope observation after HE staining, did not see inflammatory cells, erosion and ulcer.The naked eye observation of MG in colonic mucosa could see diffuse hyperemia edema, erosion and ulcer. Microscope observation after HE staining, the normal colonic mucosa structure was damaged.Besides, a lot of inflammatory cells and red blood cells could be seen. The colonic mucosa of the other 4 treatment groups were better than MG. The condition of colonic mucosa in HDG and SASP were restored the most obvious,closed to NG. The condition of colonic mucosa in MDG were restored lower than HDG and SASP.LDG were restored lower than MDG,but higher than MG. The microscope observation after HE staining was almost the same to the naked eye observation.4 Comparison of colon tissue NF-κB、COX-2 mRNA expression.The expression of NF-κB mRNA in each treatment groups were higher than NG(0.21±0.03),and lower than MG(0.58±0.04),the difference was statistically significant(P<0.05). HDG(0.29± 0.05) was lower than that of SASP(0.35±0.04), MDG(0.41±0.05) and LDG(0.50±0.06), the difference was statistically significant(P<0.05). The expression of NF-κB mRNA in SASP was lower than MDG and LDG, the difference was statistically significant(P<0.05). The expression of NF-κB mRNA in MDG was lower than LDG, the difference was statistically significant(P<0.05).All the data indicated that each treatment drugs could reduce the expression of NF-κB mRNA,and HDG was better than SASP,MDG and LDG.The expression of COX-2 mRNA in each treatment groups were higher than NG(0.33±0.06),and lower than MG(0.76±0.06),the difference was statistically significant(P<0.05). There was no statistically significant difference(P>0.05) between HDG(0.48±0.06) and SASP(0.46±0.06),and they were all lower than MDG(0.55±0.06) and LDG(0.64±0.06), the difference was statistically significant(P<0.05). The expression of COX-2 mRNA in MDG was lower than LDG, the difference was statistically significant(P<0.05). All the data indicated that each treatment drugs could reduce the expression of COX-2 mRNA.And there was no significant difference between HDG and SASP,they were all better than MDG and LDG.Conclusion:1 Xiang Lian Hua Zhuo Jie Du Decoction can improve the general conditions of UC rats, such as diarrhea and hematochezia. The Decoction also can restore hyperemia and edema of colonic mucosa and promote the restoration of erosion and ulcer. And there is a dose-effect relationship between its curative effect and dosage.2 The mechanism of Xiang Lian Hua Zhuo Jie Du Decoction to treat UC is probably likely to reduce the expression of NF-κB、COX-2 mRNA,so that lower the net of the complicated proinflammatory factor, inhibit the chain reaction of inflammatory, reduce the inflammatory reaction and the immune response of colonic mucosa, so as to achieve the purpose of treatment of UC. |
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