Effect Of Xiezhuojiedu Decoction On IL-1β、IL-10、CD14 On Ulcerative Colitis Model Rats

Objective: Ulcerative colitis is a kind of etiology and pathogenesis not very clear, to inflammation of the mucosa and submucosa of rectum and colon and extensive ulcer formation become features of chronic nonspecific inflammatory disease. A long course of the disease, lesions range, can be repeated attack, cure is difficult, seriously affect the patient’s quality of life and health of body and mind, it has been recommended to be one of the refractory diseases by World Health Organization.With the method of purging turbidity and ascending lucidity, detoxification and removing stasis, we screened an experienced Prescription of Xiezhuojiedu Decotion, which has reliable effects from many years of clinical experience. On the basis of previous studies, we established the UC rat model to study the mechanism and therapeutic effect of Xiezhuo Jiedu Decoction on UC, especially observed the effect on UC rat serum IL-1β, IL-10 and CD 14 in colonic mucosa, to provide a theoretical basis for the further clinical studies and drug use.Methods: In this experiment, 40 healthy and clean wistar Rats were taken weighing 220~250g,half male and half female,and were randomly divided into 5 groups with 8 rats in each group.They are separately named control group(BK), modeling group(MK), SASP group(LK), Xiezhuojiedu Decoction high dose group(GK), Xiezhuojiedu Decoction low dose group(DK). 2,4,6-trinitrobenzene-sulfonic acid, TNBS/ethanol combinative method of establishing model was used to Induce animal models in Rat Ulcerative Colitis 3 days after model making, BK rats were fed normally normally, MK rats were given normal saline by gavage, MK rats were given SASP by gavage, GK, DK rats were separatelygiven Xiezhuojiedu Decoction high dose and Xiezhuojiedu Decoction low dose by gavage for 14 days, observaed the curative effect and studied the mechanism of action. I studied and discussed the experiment from two aspects: Experiment 1, to observe the effect of Xiezhuojiedu Decoction on general condition and pathological effect on Ulcerative Colitis Model Rats, and determinate serum IL-1β, IL-10 content of Ulcerative Colitis Model Rats; Experiment 2, to observe the effect of Xiezhuojiedu Decoction on CD14 expression of the colonic mucosa on Ulcerative Colitis Model Rats.Results: 1 E x p e r i m e n t 1 : E f f e c t o f X i e z h u o j i e d u D e c o c t i o n o n g e n e r a l condition,pathological effect and serum IL-1, IL-10 on Ulcerative Colitis Model Rats. 1.1 The rats’ general condition and the change of weightCompared with BK, MK and other the treatment groups’ mental status, hair glossiness, appetite, weight, defecate character, ect.of the rats changed differently. The rats in GK Spirit gradually returned to normal, h a i r c o l o r a n d l u s t e r, r e s p o n d e d r a p i d l y, d i e t i n c r e a s e d, responded rapidly, the weight grew quickly, the weight、stool and other general similared to the BK group.The wight of BK(340.37±6.56)rats normal, MK(258.62±6.63) rats decreased obviously, the two groups were compared had significant difference, there was statistical significance(P<0.05). Compared with M K, the weight of GK(313.00±4.69), DK(290.75±7.61), LK(292.87± 7.58)rats increased significantly, there was statistical significance(P< 0.05). Compared with LK, the weight of GK rats increased significantly, there was statistical significance(P<0.05), there wasn’t statistical significance o n D K r a t s( P > 0. 0 5). C o m p a r e d w i t h D K, t h e w e i g h t o f G K rats increased significantly, there was statistical significance(P<0.05). 1.2 The general observation on colon and the gross score of colonic lesionsThe BK rats’ colonic mucosa were complete and continuous, light red color, smooth surface, there was no pathological changes of hyperemia, edema, ulceration. There were hyperemia, edema, erosion in the MK rats, and scattered in the ulceration, around of ulcer lesions had black necrotic tissue. There weren’t significant expansion, thickened and hyperemia, edema in the colon of GK rats, but there were scar ulcer healing, the mucosa recovery were better than DK and LK rats.Compared with the score of BK(0.362±0.164), the MK(2.965±0.372) increased significantly, there was statistical significance(P<0.05). Comp ared with MK, the score of GK(0.885±0.128), DK(1.647± 0.137), LK(1.589±0.146)rats decreased significantly, there was statistical significance(P<0.05). Compared with LK, the score of GK rats decreased significantly, there was statistical significance(P<0.05), there wasn’t statistical significance on DK rats(P>0.05). Compared with DK, the score of GK rats decreased significantly, there was statistical significance(P<0.05). 1.3 The histomorphology of rats’ colon under the light microscopeThere were different levels of congestion, edema, formed erosion and ulcer, the gland were destruction, structure were disorder, partial deletion of the epithelium, and a large number of inflammatory cells in the neutrophil infiltration. After treatment, the GK rats colonic mucosa integrity, the glands in a regular arrangement, Interstitial hyperemia and edema wasn’t obvious, local with a few inflammatory cells infiltration, abundant goblet cells,the shape was close to BK, and better than DK and LK rats. 1.4 The IL-1β of the serum on UC was detected by RIACompared with IL-1β of the serum on BK(0.108±0.010), the MK(0.205±0.009)increased significantly, there was statistical significance(P<0.05).Compared with MK, the IL-1β of the serum on GK(0.133± 0.010), DK(0.167±0.016), LK(0.169±0.009)rats increased significantly, there was statistical significance(P<0.05).Compared with LK, the IL-1β of the serum on GK rats decreased significantly, there was statistical sihnificance(P<0.05), there wasn’t statistical significance on DK rats(P>0.05) Compared with DK, the IL-1β of the serum on GK rats decreased signifyicantly, there was statistical significance(P<0.05). 1.5 The IL-10 of the serum on UC was detected by ELISACompared with IL-10 of the serum on BK(39.24±6.31),the MK(22.03±2.82) decreased significantly, there was statistical significance(P<0.05).Compared with MK, the IL-10 of the serum on GK(37.60± 2.55), DK(30.12±3.38), LK(28.98±3.90)rats increased significantly, there was statistical significance(P<0.05). Compared with LK, the IL-10 of the serum on GK rats increased significantly, there was statistical sihnificance(P<0.05), there wasn’t statistical significance on DK rats(P>0.05) Compared with DK, the IL-10 of the serum on GK rats increased signifyicantly, there was statistical significance(P<0.05). 2 Experiment 2: The CD 14 in colonic mucosa was detected by immunohistochemicalCompared with the average optical density on BK(0.078±0.035), the M K( 0. 3 4 8 ± 0. 0 5 6) i n c r e a s e d s i g n i f i c a n t l y, t h e r e w a s s t a t i s t i c a l significance(P<0.05).Compared with MK, the average optical density on GK(0.162±0.042), DK(0.246±0.047), LK(0.236±0.043)rats increased significantly, there was statistical significance(P<0.05).Compared with LK, the average optical density on GK rats decreased significantly, there was statistical sihnificance(P<0.05), there wasn’t statistical significance on DK rats(P>0.05) Compared with DK, the average optical density on GK rats decreased signifyicantly, there was statistical significance(P<0.05).Conclusion:1 The methods of using TNBS/ ethanol combined modeling method in rats with UC induced animal model was successful in this experiment.2 Xiezhuojiedu Decoction has good preponderance in treating UC, and improve the rats general condition and colonic organization form, Xiezhuojiedu Decoction high dose group is better than Xiezhuojiedu Decoction low dose group and SASP group.3 Xiezhuojiedu Decoction can significantly decrease IL-1β content on U C, i m p r o v e I L- 1 0 l e v e l o n U C; X i e z h u o j i e d u D e c o c t i o n c a n significantly decrease CD14 expression of the colonic mucosa on UC.