Experiment And Clinical Research Of The Combination Enema Of Changdiqing And Tin-like Powder On Ulcerative Colitis

Purpose:This research took rats that suffered from ulcerative colitis as investigative objects to observe the therapeutic efficacy and dose-effect relationship of ulcerative colitis by using Changdiqing and Tin-like powder to do the retention enema and influences of the two kinds of drugs to C-reaction protein(CRP) and expression of cytokines as well as the mechanism of action.As to the clinical part,we observed the therapeutic effects of coloclysis group using Changdiqing and the other group using combined Changdiqing and Tin-like powder in treating of ulcerative colitis and intended to find a therapeutic approach with benefits such as satisfactory curative effect, minor side effect,reasonable price as well as convenient direction.Method and contents:Adopted randomized control study.Experimental part:Took 80 clearing grade SD rats with equal sexual number and the weight of which between 200 to 220 grams.Numbered the rats and randomized took 10 of which out as blank group(the first group),others were built models according to combination method(2,4,6-trinitro-benzene-Sulfonic acid + alcohol).There were 14 rats died due to anesthesia accident post modeling and the rest 56 ones were randomized divided into 4 groups as follows: model group(the second group),coloclysis group with low dose of Changdiqing and Tin-like powder(the third group),coloclysis group with high dose of Changdiqing and Tin-like powder(the forth group) and control coloclysis group (the fifth group).Each group had 14 rats.Used routine forage and distilled water to feed the blank group.The other ones were begun to administrate drugs through coloclysis three days post modeling with frequency of one time per day and duration of a continuous two weeks.The blank group and modeling groups were adopting normal saline(NS) 1.0ml/100g to do the coloclysis.Coloclysis group with low dose of Changdiqing and Tin-like powder added Changdiqing 0.5ml/100g and Xileisan 0.05g/d additionally to do the colocysis.Coloclysis group with high dose of Changdiqing and Tin-like powder added Changdiqing 1.0ml/100g and Tin-like powder 0.1g/d additionally to do the colocysis.The control group chose normal saline 1ml/100g + Sulfasalazine(SASP) 0.1g/d to do the coloclysis.All the rats in each group were put to death after weighed when the research ended and got the whole colons and took 0.5cm with manifest pathological changes to do pathological classification.Meanwhile,did double antibody sandwich ABC-ELISA to detect the IL-4 level of the colon specimen. To measure CRP and IL-6 level to the separated serum from 8ml blood under storage of minus 20 degree centigrade as well.All the specimens were detected in the same batch.Clinical part:To select patients with light and midrange active stage ulcerative colitis in west medicine diagnosis and damp-heat in large intestine in traditional Chinese medicine(TCM) diagnosis as investigation objects with ages ranging from 18 to 75 years old.All the included 65 cases were randomized divided into treating group(Changdiqing combined Tin-like powder coloclysis group),which had 31cases and control group(Changdiqing coloclysis group), which had 34 cases.The former group had 17 male cases and 14 female ones, with an average age of 47.35±12.73 years old:meanwhile,the later had 18 male cases and 16 female ones,with an average age of 50.45±17.20 years old. All the cases in the two groups had no complications and symptoms beyond bowels. There were no significant differences on sex and age of the patients in the two groups(p＞0.05).To adopt 150ml Changdiqing + 1g Tin-like powder to do retention enema to the treating group with frequency of one time per day, meanwhile,to adopt 150ml Changdiqing to do retention enema to the control group with frequency of one time per day and to take one month duration as a course of treatment to the two groups.The observation items were including general data,safety surveying data and curative effect surveying data.After that,to evaluate the clinical comprehensive curative effects which could be divided as complete remission,utility and invalid.The effective powers were calculated from the percentages of the cases achieved complete remission and utility divided by the case number of each group and finally to compare whether the effective powers in the two groups had variability. Results:All the rats were suffered from symptoms and signs such as low spirit, poor appetite,diarrhea accompanied with minor blood post modeling and all the symptoms and signs were recovered in different degree during the drug coloclysis treatment.The associated results were elaborated as follows:1.Influence of Changdiqing and Tin-like powder to the intestinal mucosa pathological changes of rats with ulcerative colitis.The model group could see ulcers and plenty of inflammatory cell infiltrate with neutrophil dominance under light microscope,with a pathological lesion classification asⅣ,Ⅴdegree.Whereas,coloclysis group with low dose of Changdiqing and Tin-like powder and control group seldom could find ulcers but plenty of inflammatory cell infiltrate,with a pathological lesion classification asⅡ,Ⅲdegree and coloclysis group with high dose of Changdiqing and Tin-like powder had significantly decreased inflammatory cell infiltrate,with a pathological lesion classification asⅠ,Ⅱdegree.Each groups had statistical significance in pathological lesion classification compared to model group (P＜0.05).However,there was no statistical significance in pathological lesion classification among blank group,coloclysis group with low dose of Changdiqing and Tin-like powder,coloclysis group with high dose of Changdiqing and Tin-like powder and control group(P＞0.05).2.Influence to serum CRP(C-reaction protein) of Changdiqing and Tin-like powder to rats with ulcerative colitis.Compared to blank group,the content of CRP in model groups increased significantly with high statistical significance(P＜0.01).Comparison between model groups,the content of CRP in coloclysis group with low dose of Changdiqing and Tin-like powder, coloclysis group with high dose of Changdiqing and Tin-like powder and control group decreased significantly with high statistical significance(P＜0.01). Compared to group with low dose of Changdiqing and Tin-like powder and control group,the content of CRP in coloclysis group with high dose of Changdiqing and Tin-like powder decreased significantly with statistical significance(P＜0.05).Compared to control group,the difference of content of CRP in coloclysis group with low dose of Changdiqing and Tin-like powder had no statistical significance(P＞0.05).3.Influence to serum IL-6 of Changdiqing and Tin-like powder to rats with ulcerative colitis.Compared to blank group,the content of IL-6 in model groups increased significantly with high statistical significance(P＜0.01). Comparison between model groups,the content of IL-6 in coloclysis group with low dose of Changdiqing and Tin-like powder,coloclysis group with high dose of Changdiqing and Tin-like powder and control group decreased significantly with high statistical significance(P＜0.01) Content of IL-6 had no statistical significance among coloclysis group with low dose of Changdiqing and Tin-like powder,control group and coloclysis group with high dose of Changdiqing and Tin-like powder(P＞0.05).4.Influence to serum IL-4 of Changdiqing and Tin-like powder to rats with ulcerative colitis.Compared to blank group,the content of IL-4 in model groups decreased significantly with high statistical significance(P＜0.01). Comparison between model groups,the content of IL-4 in coloclysis group with low dose of Changdiqing and Tin-like powder,coloclysis group with high dose of Changdiqing and Tin-like powder and control group increased significantly with statistical significance(P＜0.05).Compared to group with low dose of Changdiqing and Tin-like powder and control group,the content of IL-4 in coloclysis group with high dose of Changdiqing and Tin-like powder increased significantly with statistical significance(P＜0.05).Compared to control group,the difference of content of IL-4 in coloclysis group with low dose of Changdiqing and Tin-like powder had no statistical significance(P＞0.05).There was no severe adverse effect detected in the two groups after the ending of clinical trail,which had suggested the patients compliance was satisfactory and there was no amotic case as well.We took the clinic symptom recover and colonoscopy finding on rechecking as curative effect assessment standards and the results of clinical therapeutic effect of the two groups post treatment were as follows:among the 31 cases in the treating group, 11 cases complete remission,17 utility and 3 invalid,with an in total effective rate of 90.32%:among the 34 cases in the control group,6 cases complete remission,18 utility and 10 invalid,with an in total effective rate of 70.59%.The two sets had statistical significance(P=0.0470,P＜0.05) which had suggested that clinical curative effect in the treating group was superior than the control group in ulcerative colitis.Conclusions:In this research,we adopted combination method(2,4,6-trinitro-benzene-sulfonic acid + alcohol) to cope the ulcerative colitis animal model,and could find ulcers and plenty of inflammatory cell infiltrate with neutrophil dominance under light microscope,with a pathological lesion classification asⅣ,Ⅴdegree.Meanwhile,the content of CRP and IL-6 were more than common ones while IL-4 lower than common ones(P＜0.01),which had suggested the occurrence of ulcerative colitis built through combination method might have relationship with high expression of CRP and IL-6.In another groups,we adopted Changdiqing(a kind of traditional Chinese medicine),Changdiqing and Tin-like powder with different dosage to do coloclysis and Sulfasalazine to do coloclysis and to investigate their pharmacologic action.We found all the drugs had a certain anti-ulcer effect which could release symptoms of ulcerative colitis in the animal models and soften colonic ulcer and inflammatory pathological changes.Among the treating groups,coloclysis group with high dose of Changdiqing and Tin-like powder had best curative effect by significantly improving the pathogenic classification of the rats with ulcerative colitis,decreasing the serum content of CRF and IL-6 and increasing serum content of anti-inflammatory factor IL-4,which might be one of the pharmacologic mechanism of Changdiqing and Tin-like powder.In clinic practice,ulcerative colitis were mainly started in rectum and the prevalence of rectum and sigmoid colon reached as many as 70%.Meanwhile, course of disease of ulcerative colitis in our country were mainly appeared to be recurrent attacks with comparatively gently pathogenetic conditions of minor and midrange and the extent of diseases were mainly located in left hemicolon.For the reasons above,using traditional Chinese medicine to do retention enema had considerable advantages.On the basis of prophase work, we chose Changdiqing and Tin-like powder to do the coloclysis and compared the effect to group just using Changdiqing only.Results had suggested both the two treating method had no severe adverse effect and the patient had satisfactory compliance to this way as well.However,group using Changdiqing and Tin-like powder to do the coloclysis had better clinical curative effect had had other benefits such as minor side effects,strong compliance, reasonable price,which could be easily widely accepted by patients suffered from ulcerative colitis and solve their sufferings.The prostecdtive efficacy and precisely treating mechanism still needed to be investigated.