| Objective:This subject studied treatment of effect and mechanism of Dongyao Wuwei Zhixie decoction by making rat model of UC. In particular, we use Dongyao Wuwei Zhixie decoction to treat UC rats and then we observe the content is Ultrastructure of colonic mucosa and content of cytokines (IL-1, IL-6, IL-4, IL-10) which come from serum of the UC rats. We also observe other indicators such as:the expression of nuclear factor kappa B p65and Gene Expression of TLR2and TLR4in colon in mucosa of rats with ulcerative colitis.To deeply research Dongyao Wuwei Zhixie decoction to immunity action mechanism of UC rats. in order to reach the level of gene expression for the national medicine treatment of UC mechanism. To provide the experimental basis and theoretical basis for anti UC Dong drug treatment of UC with high quality.Methods:In the experiment, the60SD rats (SPF, half male and half female) were randomly divided into normal group, model group, beautiful salad diazoxide (0.5g.kg-1) group and Dong five drugs Zhixie Decoction low, middle and high dose group,(5.5,11,22g. kg-1) were divided into6groups, each group of10only. Except the normal group, the other5groups were prepared the model of UC rat through the5%2,4,6-three TNBS and Ethanol Compound enema. On the third day after modeling, The normal group and the Pathologic control group was administered with equal volume of distilled water, at the same time, the Dongyao Wuwei Zhixie Decoction low, medium and high dose group, Mesalazine group were given medicine by respectively according to the corresponding dose,1times a day, for10consecutive days. At the end of the treatment, the changes of pathology and ultrastructure of colonic mucosa of colon were observed by the light microscope and the electron microscope respectively,such as the content serum of IL-1, IL-6and IL-4, IL-10in every group of rats were detected by ELISA.The expression of NF-κ B p65in colon tissue were detected by Immunohistochemical staining(Envision method). The gene expression of TLR2, TLR4in colon tissue were also detected by Application of real-time fluorescent quantitative PCR (real-time PCR). The results of observation will be given statistics and analysis.Results:The results showed:①The changes in symptoms of rats in the Pathologic group can be observed very poor. The rats in the model group can been observed the changes of the symptoms such as intermittent diarrhea with mucus and blood at stool, emaciation, laziness, no luster in hair, poor appetite, loss of weight. The weight of rats in the Pathologic group was significantly lower than that in the normal group(P<0.01). The Pathologic group was observed different size of the ulcer in the intestinal mucosa, under the light microscope, inflammation, edema and ultrastructure damage is very serious. The colon tissue damaged score in pathologic group is higher than that in normal group(P<0.01). After treatment, their symptoms and sighs obviously improved. The colon tissue damaged score in pathologic group was higher than that in the high, medium, low dose group of Dongyao Wuwei Zhixie decoction and Mesalazine group. The high, medium dose group of Dongyao Wuwei Zhixie decoction were significantly better than the Pathologic group in the degree of colonic mucosal inflammation, hyperemia and ultrastructure.②The level of IL-1, IL-6in pathologic group was significantly higher than that in normal group, and The level of IL-4、 IL-10was significantly lower than that of the normal group, tow groups have very significant difference(P<0.01). After treatment, the level of IL-1、IL-6in the high, medium, low dose group of Dongyao Wuwei Zhixie decoction and Mesalazine group was lower than in pathologic group (P<0.01or P<0.05), and the level of IL-4、IL-10was significantly higher than that in pathologic group(P<0.01or P<0.05).③The expression of NF-κB p65in pathologic group were significantly higher than that in normal group, tow groups have very significant difference(P<0.01). After treatment, the expression of NF-κB p65in Dongyao Wuwei Zhixie Decoction high, medium dose group and Mesalazine group was significantly lower than that in pathologic group, every groups have very significant difference to that in pathologic group (P<0.01).④The gene expression of TLR2mRNA、TLR4mRNA in pathologic group were significantly higher than that in normal group, tow groups have very significant difference(P<0.01). After treatment, The gene expression of TLR2mRNA、TLR4mRNA in high, medium dose group of Dongyao Wuwei Zhixie Decoction and Mesalazine group was significantly lower than that in pathologic group, every groups have very significant difference(P<0.01).The gene expression of TLR2mRNA、TLR4mRNA in low dose group of Dongyao Wuwei Zhixie Decoction have not significant difference to that in pathologic group(P>0.05).Conclusion:The above result indicated that this experiment UC rat model duplicates successfully.Every groups of Dongyao Wuwei Zhixie Decoction Have a good therapeutic effect on TNBS/alcohol UC rat. In particular, it is better effect in high dose group of Dongyao Wuwei Zhixie Decoction, the treatment effect is related to the size of dose. There is a certain relationship between dose-effect. Dongyao Wuwei Zhixie Decoction could reduce the content of pro-inflammatory cytokines IL-1、IL-6, and improve the level of pro-inflammatory cytokines IL-4、IL-10, Control the inflammatory response, in general can regulate the balance of cytokine network. Nuclear factor-kappa B play an important control role in the pathogenesis of the UC. The high expression of TLR2, TLR4in colon tissues was closely related with with the pathogenesis of the UC, Dongyao Wuwei Zhixie Decoction could inhibit the activation of NF-κB, and decrease the expression of TLR2mRNA、TLR4mRNA, then it could inhibit the activation of TLRs/NF-κB.Regulation of inflammatory mediator release, and thus play a role in the regulation of cell factor, it could effectively inhibit the inflammatory reaction of UC, promote the recovery of immune function and play a role in the treatment of UC. This may be an important mechanism of immune regulation which Dongyao Wuwei Zhixie Decoction could treat the FNBS type of UC rat. |
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