Effect Of Xiezhuojiedu Decoction On IL-8,IκB-α And TLR4 On Ulcerative Colitis Model Rats

Objective:According to the pathogenesis of Ulcerative Colitis(UC),we composed Xiezhuojiedu Decoction,the effect of which is purging turbidity, ascending Lucidity, detoxifying and removing blood stasis. We carried out the animal experiment to explore the effect of Xiezhuojiedu Decoction on IL-8,IκB-α,TLR4 on Ulcerative Colitis Model Rats,and compare the therap -eutic effects with sulfasalazine tablets(SASP), elucidate the possible mech- anism of this decoction on Ulcerative Colitis ,so as to provide new ideas for the treatment of Ulcerative Colitis.Methods:2,4,6-trinitrobenzene-sulfonic acid ,TNBS/ethanol combinative method of establishing model used to Induce animal models in Rat Ulcerative Colitis in this study, 45 healthy and clean wistar Rats were taken ,which were male, weighting 220--250g.8 rats were randomly chosen to be the control group, the other rats were given 30% Ethanol Solution which contains 30mg TNBS, to make Ulcerative Colitis animal models. Three days later,5 rats were randomly chosen to be killed to take their colon models. If there are hype -remias, edemas and ulcers, the UC rats model making can be thought to be successful. The other rats were randomly divided into 4 groups with 8 rats in each group, and they are separately named modeling group,SASP group, Xiezhuojiedu Decoction high dose group,Xiezhuojiedu Decoction low dose group. The control group,modeling group rats were given normal saline by gavage.The rats of SASP group,Xiezhuojiedu Decoction low dose group, Xiezhuojiedu Decoction high dose group were separately given SASP at 0.3g/kg, water decoction of Xiezhuojiedu Decoction at 10.0g/kg and 20.0g/kg by gavage ,Once a day for 14 days and observed the effect.I studied and discussed the experiment from three aspects: Experiment 1, to observe the effect of Xiezhuojiedu Decoction on general condition, pathological effect and IL-8 in serum on Ulcerative Colitis Model Rats; Experiment 2, to observe the effect of Xiezhuojiedu Decoction on the expression of IκB-αon Ulcerative Colitis Model Rats'colon; Experiment 3, to observe the effect of Xiezhuo jiedu Decoction on TLR4mRNA and TLR4 protein expression of the colonic mucosa on Ulcerative Colitis Model Rats separately by RT-PCR and immuno- histochemistry. Using SPSS 13.0 for statistical analysis.The main object of this study is to elucidate the possible mechanism of Xiezhuojiedu Decoction on Ulcerative Colitis through the animal experiment.Results:1 Experment 1 showed :Compared with the modeling group, the rats'general condition changed differently in modeling group and all the treatment groups comparing with the control group. All those mental status, activity condition, hair glossiness, appetite, weight defecate character,ect. were better in Xiezhuojiedu Decoction high dose group than the other groups,model group was the worst.Pathological damage:Observed under the microscope,the modeling group rats'mucous layer deleted, congestion and edema,inflammatory cell infil- tration were found in inflammatory cell infiltration,ulcers and fibrosis could be found in some of them; ulcers were superficial, congestion ,edema and less inflammatory cell infiltration in SASP group and Xiezhuojiedu Decoction low dose group; the mucous layer was complete and there were almost no congestion and edema,inflammatory cell infiltration in Xiezhuojiedu Decoc- tion high dose group.The scores of histopathological damage : there were not damages on the control group rats'colon , The scores of histopathological damage increased in modeling group,compared with the modeling group(6.375±1.188),The scores of histopathological damage decreased significantly in SASP group (4.125±1.356), Xiezhuojiedu Decoction high dose group(1.625±1.060) and Xiezhuojiedu Decoction low dose group(4.250±1.282),all had significant differrence (P<0.01). compared with SASP group , Xiezhuojiedu Decoction low dose group didn't have significant difference(P>0.05); the scores of histopathological damage in Xiezhuojiedu Decoction high dose group were better than SASP group and Xiezhuojiedu Decoction low dose group, had significant difference (P<0.01).IL-8 in serum on Ulcerative Colitis Model Rats: Compared with control group(0.310±0.161), IL-8 in serum in modeling group (0.523±0.698) rose obviously ,had significant difference (P<0.01); SASP group(0.389±0.118), Xiezhuojiedu Decoction low dose group(0.397±0.082) and Xiezhuo- jiedu Decoction high dose group(0.259±0.149) did not have significant difference (P>0.05). Compared with modeling group,IL-8 in serum in SASP group, Xiezhuojiedu Decoction high dose group and Xiezhuojiedu Decoction low dose group deceased significantly, had significant difference (P<0.01). Compared with SASP group, IL-8 in serum in Xiezhuojiedu Decoction high dose group deceased significantly, had significant difference (P<0.05); Xiezhuojiedu Decoction low dose group did not have significant difference (P>0.05).Compared with Xiezhuojiedu Decoction low dose group,IL-8 in serum in Xiezhuojiedu Decoction high dose group deceased significantly, had significant difference (P<0.05).2 Experiment 2 showed :The expression of IκB-αprotein on Ulcerative Colitis Model Rats'colon:Compared with control group(12.500±3.251), the expression of IκB-αprotein in modeling group (49.750±8.137) rose obviously,had significant difference (P<0.01); Compared with modeling group, the expression of IκB-αprotein in SASP group ,Xiezhuojiedu Decoction high dose group(17.125±5.718) and Xiezhuojiedu Decoction low dose group decreased significantly, had significant difference (P<0.01). Compared with SASP group(23.875±7.415), the expression of IκB-αprotein in Xiezhuojiedu Decoction high dose group(17.125±5.718) was lower than SASP group,had significant difference (P<0.01); Xiezhuojiedu Decoction low dose group(28.000±4.567) did not have significant difference (P>0.05). Compared with Xiezhuojiedu Decoction low dose group, the expression of IκB-αprotein in Xiezhuojiedu Decoction high dose group was lower than SASP group ,had significant difference (P< 0.01).3 Experiment 3 showed :TLR4 protein expression in the colonic mucosa:Compared with control group(13.500±2.449), TLR4 protein expression in modeling group(57.250±4.166) rose obviously ,had significant difference (P<0.01). Compared with modeling group, TLR4 protein expression in SASP group(37.125±3.227), Xiezhuojiedu Decoction low dose group (35.375±3.623) and Xiezhuojiedu Decoction high dose group(17.625±2.066)deceased significantly,had signi- ficant difference (P<0.01). Compared with SASP group, TLR4 protein expressionXiezhuojiedu Decoction high dose group decreased significantly, had significant difference (P<0.01). Xiezhuojiedu Decoction low dose group did not have significant difference (P>0.05). the expression of TLR4 protein was lower than Xiezhuojiedu Decoction low dose group, had significant difference (P<0.01).The expression of TLR4mRNA in the colonic mucosa : Compared with control group(0.181±0.016),the expression of TLR4mRNA in modeling group(0.519±0.043)rose obviously ,had significant difference (P<0.01). Compared with modeling group, the expression of TLR4mRNA in SASP group(0.316±0.016),Xiezhuojiedu Decoction low dose group(0.343±0.028) and Xiezhuojiedu Decoction high dose group(0.149±0.037) deceased signi- ficantly, had significant difference (P<0.01). Compared with SASP group, the expression of TLR4mRNA in Xiezhuojiedu Decoction high dose group deceased significantly, had significant difference (P<0.01); Xiezhuojiedu Decoction low dose group did not have significant difference (P>0.05). The expression of TLR4mRNA in Xiezhuojiedu Decoction high dose group was lower than Xiezhuojiedu Decoction low dose group, had significant difference (P<0.01).Conclusion:During the long clinical observation and study, we proposed turbidity toxin theory: turbidity toxin stasis , lipid membranes and su -perficial venules injury, blood and meat rot , block and suppurate;after a long time , turbidity toxin is cohesive , intestinal conduction loses control and then this disease occurres . And compoed Xiezhuojiedu Decoction the function of which is purging turbidity and ascending lucidity, detoxification and removing stasis,it has reliable effects.Now we made Ulcer- ative Colitis Models to obse rve the change of pre and post treatment. The research results showed that: Xi- ezhuojiedu Decoction can improve the rats'general condition and the scores of histopathological damage, decrease IL-8 in serum, the expression of IκB-αpro -tein and TLR4 cell expression rate, the expression of TLR4mRNA in the co- lonic mucosa,consequently inhibit infla- mmatory cells,repair the damaged in- testinal mucosa, relieve the infla-mmation, accelerate the ulcer healing. The re -sults proved the effect of Chinese medicine Xiezhuojiedu Decoction on Ulcer ative Colitis Model Rats , the therapeutic mechanism is possibaly related to its influence on some inflam -matory factors.