| Objective:Ulcerative Colitis is a kind of chronic nonspecific colitis disease which specially occurs in the colon and mainly infiltrates mucosa and submucosa.This disease belongs to“diarrhea"areas in traditional Chinese medicine.The etiology and pathogenesis of this disease is not very clear,which relates to susceptible genes,environmental factors and immune system abnormalities.But more and more studies have shown that the disease is related to the relationship between intestinal symbiotic bacteria and intestinal epithelial barrier and the intestinal mucosal immune imbalance it drives.In recent years,the studies of the treatment aimed at intestinal lymphocyte homing mechanism have some progress,but serious adverse reactions these drugs caused make the experiment had to stop.So exploring the traditional Chinese medicine preparations of high efficacy and low toxicity becomes a research hotspot.Previous studies of the experiment show that the croton cream and rhubarb and its main ingredients Emodin,Gallic Acid respectively compatibility reduce the pathological damage of UC intestinal mucosa through different targets to promote damage repair.The drug has a good pharmacological effect in the treatment of UC rats.Rhubarb polysaccharide is one of the main water-soluble components of rhubarb,has a variety of biological activity and plays an important role in the drug activity of rhubarb.In order to fully explain the effect of rhubarb and croton cream compatibility on the immune system function of intestinal mucosa,this study intends to take the intestinal mucosal immune barrier and the lesion process of intestinal lymphocyte homing in the UC rats as targets though the different doses of rhubarb polysaccharide and croton cream compatibility to explores the mechanism whether the compatibility in the way of intervening intestinal lymphocytes homing and participating in the intestinal mucosal immune barrier protection or regulation in the UC rats achieves the"antidiarrheal"effect.Methods:1.Clean-grade wistar male rats with TNBS+50%ethanol enema to establish stable UC rats models.60 rats were randomly divided into normal control group(N),model control group(C),sulfasalazine group(Y),and croton cream+three doses of rhubarb polysaccharide group(RHP)as high group(B+R80),medium group(B+R60)Low group(B+R40),totally6 groups and each group of 10.Three days after modeling,the corresponding drugs were given to each group orally.2.Using Disease activity index(DAI),colon mucosa damage index(CMDI),histopathology and organ index as indicators to evaluate the success rate of the model and to observe the effect of drug intervention preliminarily.3.Seven days after continuous administration,sacrificing the rats to collect anticoagulant whole blood and related tissue samples.CD3~+,CD4~+,CD8~+and CD4~+/CD8~+T lymphocytes in peripheral blood and colonic mucosa of UC rats were detected by flow cytometry.The expression of MAdCAM-1 mRNA in colonic mucosal pathology was detected by RT-PCR.The expressions of?L?2 and ICAM-1 in colonic tissues were detected by immunohistochemistry.Results:1.In terms of gross morphology and tissues and organs,the scores of DAI,CMDI and HS in group C were significantly higher than those in group N(P<0.01),while the thymus index was significantly decreased(P<0.01).Compared with the model group,the DAI and CMDI scores of the drug given groups showed a significant decrease(P<0.05).The HS scores were decreased in different degrees,the difference was statistically significant(P<0.01).There was no significant dose-effect relationship between the three dose groups,but the DAI,CMDI and HS scores were significantly decreased in group B+R40.2.The flow cytometry results showed that the number of CD3+and CD8+in peripheral blood T lymphocyte subsets was significantly lower than that in group N(P<0.01,P<0.05),while CD4+,CD4+/CD8+ratio was significantly higher than that in group N(P<0.05,P<0.01).Compared with the group C,the level of CD8+increased and CD4+/CD8+ratio decreased in the drug given groups,and the effect of group B+R40 was the most significant.The number of CD4+and the CD4+/CD8+ratio in intestinal mucosa of group C decreased compared with group N(P<0.01),which recovered after drug intervention,the effect of group B+R40 was the most significant.3.Semi-quantitative RT-PCR results showed that the expression of MAdCAM-1mRNA in model group was significantly higher than that in group N,and the expression of MAdCAM-1 in each group was on a downward trend after treatment,the expression of MAdCAM-1 in group B+R60,group B+R40 was similar to that of blank control group(P>0.05).Comparing each dose group,the group B+R60,group B+R40 were better than that of group B+R80,and the B+R combination group was more effective than positive drug group in lowering the expression level of MAdCAM-1.4.Immunohisto chemistry results showed that the positive expression of?L?2and ICAM-1 in group C was significantly higher than that in group N(P<0.01).Compared with group C,the positive expression of ICAM-1 in B+R three dose groups was decreased intentionally,and the expression of?L?2 in group B+R80,groupB+R40 was decreased intentionally.Conclusions:1.The crotoncream and rhubarb polysaccharide combination can regulate the disorderly immune function;reduce intestinal inflammation by improving the proportion of T lymphocyte subsets in the ulcerative colitis rats.2.All the crotoncream and three doses of rhubarb polysaccharide groups can in hibitlymphocyte homing indifferent degrees,reduce the inflammatory response,promote mucosal repair by reducing the expression of specific adhesion molecules.Among them,the low dose groupwas the most significant.3.The crotoncream and rhubarb polysaccharide combinationis one of the effective material bases of the couplet medicinesof rhubarb and croton cream in the treatment of ulcerative colitis. |
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