Study On Clinical Effects And Animal Experiment Of Guangchangfang In Treating Ulcerative Colitis

ObjectiveUlcerative colitis is a disease of digestive tract characterised by chronic inflammation and ulceration of colonic mucous membrane. The inflammation of colon is chronic and nonspecific. The clinical manifestation of UC comprise mucus, pus and blood stool, diarrhea, abdominal pain, tenesmus and so on. UC usually last a very long period and do easy to relapse. The pathogenesis of ulcerative colitis is not very clear. Immune factors, environment, microorganism, heredity may have relationship with it. Among all of above, immune factors have been seriously concerned. At present, the incidence of ulcerative colitis is more and more rising. Sulfasalazine and mesalazine slow release tablets are primarily healing medicine in western medical science. In active stage of ulcerative colitis, these medicine can quickly control the symptoms. The clinical remission rate is high. But the disease is easily recur after drug withdrawal. In addition, the side effects of western medicine restrict its clinical using. Traditional Chinese Medicine shows its superiority in improving quality of life, lowering recurrence rate and lower spend. Guanchangfang is a proved recipe of guang'an men hospital, china academy of Chinese medical sciences. The curative effect of guanchangfang is positive which is proved in last curing patient. Based on the theory of Traditional Chinese Medicine, the effect of guanchangfang are dispelling dampness, heating-clearing and detoxicating, spasmolysis and alleviating pain. The topic evaluate its therapeutic effect. Besides, it investigates the mechanism of restraining the reaction of inflammatory injury, promoting the healing of colonic mucous membrane in the use of animal experiment. We analyze the mechanism of guanchangfang in order to offer an objective method and medicine to cure ulcerative colitis.MethodsThe clinical research study was strictly designed under correlated standard. The random positive control method was adopted, on the basis of diagnosis criterion established on Inflammatory Bowel Disease Seminar at chengdu in 2000 year.40 cases of ulcerative colitis are selected from out-patient clinic (OPD) of Qilu hospital of Shandong university. Treated group (20 cases) were received guanchangfang(coptis chinensis 10g, forsythia suspense 12g, jasmine fruit 10g, ledebourellae 12g, paeoniaealbe 12g, glycyrrhizae 10g). Control group were received sulfasalazine (2g/d). Both treatment course was 3 weeks. Mainly observed items:1. Therapeutic effect, including clinical general effect, change of Traditional Chinese Medicine syndrome.2. The serum levels of D-dimer (D-D),Platelet (PLT) and Mean Platelet Volume (MPV).In animal experiment, the SD rats were randomly divided into 6 groups:normal group, model group, treated group (low dose), treated group (middle dose) treated group (high dose) and mesalazine group. The last 5 groups are UC rats induced by 2,4,6—trinitrobenzenesulfonic Acid(TNBS)/ethanol mixture. The treatment time is 18 days, then we took serum and colon tissues from the rats. Mainly observed items:1. The effects of the general behaviors, living state and the weight of the rats.2. The pathological change of colon of rats both gross and microscopic observations.3. Serum level of Interleukin-4(IL-4), Interleukin-10(IL-10).4. The mRNA expression of peroxisome proliferators activated receptorγ(PPARγ), nuclear factor kB p 65(NF-kB p 65), Toll-Like Receptors 2(TLR 2)from colon mucous membrane were detected by reverse transcription PCR (RT-PCR).Results1. Evaluation of therapeutic effect of clinical research:(1) Clinical therapeutic effect of treated group was better than control group. The total effective rate of treated group was 90.00% and control group was 75.00%. The statistic significance was difference. Treated group was obviously better than control group (P<0.05).(2) The serum D-D and PLT of treated group were lower than control group. The statistic significance was difference (P<0.05). The serum MPV of treated group was remarkablely higher control group. The statistic significance was difference (P<0.05).(3) Adverse effect rating of treated group was 5% and that of control group was 10%. The statistic significance was not difference (P>0.05).2. Animal experiment research:(1) On the aspect of the general behaviors, glossiness of coat and improving the living state, treated group and mesalazine group were better than model group. Besides, treated group (low dose) was worse than other treated group and mesalazine. Maybe the effect of medicine was concerned with the dose of medicine. The weight of all the rats was not significant difference. In addition, on the aspect of pathological change of colon of rats both gross and microscopic observations, treated group and mesalazine group were better than model group. Treated group (middle and high dose) and mesalazine group were better than treated group (low dose)(2)Serum IL-4 of all groups was lower than normal group (P<0.05). Serum IL-4 of treated groups and mesalazine group were higher than model group (P<0.05) Treated group (middle and high dose) and mesalazine group were higher than treated group (low dose) (P<0.05). Treated group (middle and high dose) and mesalazine group were not significant difference(P>0.05).(3) Serum IL-10 of all groups was lower than normal group (P<0.05). Serum IL-10 of treated groups and mesalazine group were higher than model group (P<0.05) Treated group (middle and high dose) and mesalazine group were higher than treated group (low dose) (P<0.05). Treated group (middle and high dose) and mesalazine group were not significant difference(P>0.05). Guanchangfang could increase the level of IL-4 which could improve the inflammation of colon.(4) The colon mucous peroxisome proliferators—activated receptorγmRNA of normal group was significantly higher than other groups (P<0.05). The colon mucous peroxisome proliferators—activated receptorγmRNA of treated group (middle and high dose) and mesalazine group were higher than model group and treated group (low dose) (P<0.05). The colon mucous peroxisome proliferators—activated receptorγmRNA of treated group (low dose) was similar with model group (P>0.05). Maybe the effect of guanchangfang is elevation to the level of peroxisome proliferators—activated receptorγmRNA which could inhibit the activation of NF-κB. So the express of proinflammatory factor was interrupted. The development and growth of inflammation was prevented. The role of guanchangfang was similar with excitomotor of peroxisome proliferators activated receptorγ.(5) The colon mucous nuclear factor k B p65 mRNA of model group, treated group and mesalazine group were significantly higher than normal groups (P<0.05) That of treated group and mesalazine group were lower model group (P<0.05) In addition, the colon mucous nuclear factorκB p65 mRNA of treated group (low dose) was higher than treated group (high and middle dose). Guanchangfang could cut down the level of nuclear factorκB p65 mRNA which could prevent the activation of proinflammatory factor. The deterioration of endothelial cell was improved and some ulcerative colitis was cured.(6) The colon mucous Toll-Like Receptors 2 mRNA of model group, treated group and mesalazine group were significantly higher than normal groups (P<0.05) That of treated group and mesalazine group were lower model group (P<0.05) In addition, the colon mucous Toll-Like Receptors 2 mRNA of treated group (low dose) was higher than treated group (high and middle dose). Guanchangfang probably could control the gene expression of Toll-Like Receptors 2 and down-regulate the activation of nuclear factorκB which could block the signaling pathway of proinflammatory cytokines. The excessive immune response was influenced that maked endothelial cells restored. In addition, guanchangfang produce a marked effect in a dose-dependent manner. Above routes could be the mechanism that guanchangfang could improve the ulcerative colitis.ConclusionGuanchangfang is effective to treat ulcerative colitis whose type of syndrome is endoretention of damp heat by using the methods of clearing away heat evil, promoting diuresis and spasmolysis. The therapeutic effect is obviously better than SASP. It shows its superiority Compared with western medicine and has not obvious ill-effect. It can lower the serum content of D-dimer and platelet, and can increase mean platelet volume in sera, which may be an important pathway in clinical trail. The following is the mechanism of guanchangfang:1. Raising the expression of IL-4 and IL-10, preventing the aggregation of inflamed cell, helping the balance between proinflammatory factor and anti-inflammatory factor, restrainting the idiopathetic immune reaction.2. Raising the expression of colon mucous peroxisome proliferators—activated receptorγ, preventing nuclear factor k B and toll-Like receptors 2 so as to block the signaling pathway of proinflammatory cytokines, cut down the condition of local immunity, enhance mucosa epithelial cell proliferation and promote the repair of injuried mucous membrane.