| Objectives:Though the observation of the expression of LanYinFeng YangHuaZhuoJie Du Decoction effects on Ulcerative Colitis(UC) model rats and its impact on the rats'general situation,DAI scores,serum tumor necrosis factor-α(TNF-α) levels, colon tissue Toll-like receptor 4 (TLR4) receptor and Nuclear transcription factor-Kappa Bp65(NF-kBp65) protein, and compared with sulfasalazine (SASP) group, try to further confirm that the Decoction's effect and mechanism of UC from an experimental point of view, explore and research new treatment methods and effective drugs, in order to provide an experimental basis for clinical and broaden the therapeutic areas of UC.Methods: 60 eight-week-old healthy male wistar rats were randomly divided into 5 groups,each group 12 rats,they were respectively the normal group(NG),model group(MG),high dose group of traditional Chinese medicine(HDG),low dose group of traditional Chinese medicine(LDG) and the SASP group(SPAP).Using trini-trobenzenesul-fonic acid/ethanol (TNBs/ethanol)in rats induced by modeling the joint UC animal models (MK).On the 3th day,after the success of MK,and than respectively give the HDG,the LDG and the SASP gavage treatment,give the NG the same water. Record the disease activity index (DAI) score of Rats in each group daily.Rats in each group were sacrificed after 14th day of gavage treatment, each rat 4ml of blood were taken to do the TNF-αlevels detection ,collect the colon specimen immediately and fix them in the 10% formalin for more than 24 hours, Gradient ethanol dehydration,paraffin-embedded, conventional slices (thickness 3um),HE staining, observe the colon specimen under the microscope,some sections are used for the Immunohistochemical analysis of the expression of TLR4 and NF-kBp65. Results:1. The general situation and DAI scores of each group rats:Compared with the NG, the general situation of rats lacked of energy, activity decreased extrados, hair loose, dull yellowish fur, decreased appetite, bloody diarrhea, or mucus after modeling,in which the MG rats decreased the most. At the end of the experiment,the general situation of rats in each group all had different levels of improvement in general condition, in which the MG rats recovered the worst.The general condition of the HDG rats recovered the most ,which were close to the NG, while the state and recovery of the SPAP and the LGD of rats was between the HDG and the NG, there was no significant difference between the two groups.In the DAI scores: compared with the NG ,the MG (2.56±0.23), the HDG (0.68±0.13), the LDG (1.08±0.11) and the SASP (1.09±0.10) p <0.05,statistically significant;the HDG (0.68±0.13) compared with the LDG (1.08±0.11) and the SASP (1.09±0.10) p <0.05, statistically significant;the LDG (1.08±0.11) compared with the SPAP (1.09±0.10) p>0.05,no statistical significance.2. the general observation of colonic mucosa and the histopathological concept:The MG was the worst by the visual observation of rat's colon mucosa,while the HDG was much better, closely to the NG.The LDG and The SPAP was between the HDG and the NG, there was no significant difference between the two groups; With the histopathologic aspects: compared with the NG, the intestinal epithelial cell junction integrity of degrees, the gland structure, and goblet cells of rats in each group after modeling, had all changed in different levels, we could see different levels of congestion, edema, erosion and ulceration of lesion formation and the MG was most apparent group; at the end of the experiment, the rats'colon mucosa of each treatment group all recovered in different levels ,in which the degree of the HDG recovered well, closely to the NG.There was still visible edema and congestion model group, inflammatory cell infiltration and ulceration of the existence in the MG,which recovered poorly. The LDG and The SPAP was between the HDG and the NG, there was no significant difference between the two groups.3. Effect of LanYinFengYangHuaZhuoJieDu Decoction on UC Model Rats'serum TNF-αcontentCompared with the MG (1.10±0.29), the serum TNF-αlevels of each treatment group was lower p <0.05, statistically significant; the HDG (0.75±0.10) compare with the SPAP (0.90±0.07)and the LDG (0.88±0.09) p <0.05, statistically significant; Comparison between the LDG (0.88±0.09) and the SPAP (0.90±0.07) p> 0.05, no statistical significance.4. Expression of LanYinFengYangHuaZhuoJieDu Decoction on UC Model Rats'NF-kBp65 and TLR4Compared with the MG(194.2±12.4,81.6±8.6) , the expression of NF-kBp65 and TLR4 in other groups p <0.05, statistically significant; the HDG (89.0±10.7,7.6±2.4) compare with the SPAP (103.5±9.47,36.4±7.7)and the LDG (105.0±10.2,46.0±8.9) p <0.05, statistically significant; Comparison between the LDG (105.0±10.2,46.0±8.9)and the SPAP(103.5±9.47,36.4±7.7) p> 0.05,no statistical significance.Conclusions:1. The method of TNBs / ethanol can be successfully induced rat model of UC, the clinical manifestations and pathological performance with human UC have a high degree of similarity. The model method is simple, affordable and repeatable.2. LanYinFengYangHuaZhuoJieDu Decoction can not only rapidly alleviate the UC rats'diarrhea, mucous, blood stool and other clinical manifestations but also well control the organization congestion, edema, erosion and ulcers to promote healing of the organization.A dose-dependent efficacy of traditional Chinese medicine is related.3 Mechanism of LanYinFengYangHuaZhuoJieDu Decoction on UC Model Rats might be: 1) Using TNF-αand other pro-inflammatory cytokines to regulate TLRs / NF-kB pathway, inhibit NF-kB'activation and release, than reduce the inflammation and adjust the body's immunity. |
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