The Study On Effect Mechanism Of Ulcerative Colitis Rat Intestinal Mucosal Injury By Intervention Of "Ni Liu Wan Zhou"

Objective: Based on the theory of "Ni liu wan zhou",this study explored the effects of Ginseng Baidu Powder on the content of cytokines,gastrointestinal function and the expression of tight junction protein in intestinal mucosal barrier in rats with ulcerative colitis,and provided experimental basis for the treatment of ulcerative colitis by the theory of "Ni liu wan zhou" and Ginseng Baidu Powder.Methods: SD rats were induced by TNBS to establish ulcerative colitis model.Treatment of Ginseng Baidu Powder by Gastric Irrigation.To observe the general situation of rats;to detect gastrointestinal function;and observe the histopathological changes of colon under light microscopy.The content of MDA was determined by thiobarbituric acid reaction method.The activity of SOD was determined by xanthine oxidase method.The activity of MPO was determined by colorimetry.Detection of serum levels of IL-1beta,IL-6,TNF-alpha and IFN-gamma in rats by ELISA.The expression of tight junction proteins Occludin,Claudin-5 and ZO-1 in colon tissues were detected by real-time fluorescence quantitative PCR and Western Blot immunoblotting.Results:(1)General state and DAI score: Compared with the model group,the mental state,food intake,skin color,fecal mucus and pus blood of SD rats in the positive control group,the high,middle and low dose groups of Ginseng Baidu Powder were significantly improved.Compared with the model group,the weight of rats in the middle dose group and the low dose group of Ginseng Baidu Powder increased significantly(P<0.05).DAI score: The scores of model group increased significantly,while those of positive control group and Ginseng Baidu Powder groups decreased(P<0.05).(2)Observation: in model group,the colon mucosa became congested and edema,thickened and coarsened,and the length became shorter.In addition,intestinal adhesion was obvious,erosion and ulcer were formed.The colonic mucosa of rats in the positive control group and the high,middle and low dose groups of Ginseng Baidu Powder were repaired to a certain extent,and the ulcer area,the weight and length of the colon were improved.CMDI score: Compared with model group,there was significant difference in colon length between positive control group and high,medium and low dose group of Ginseng Baidu Powder(P<0.05).There was no significant difference in colon weight,colon index,ulcer area and CDMI score between positive control group and high,middle and low dose groups of Ginseng Baidu Powder(P>0.05).Histopathological observation under HE staining microscope: In model group,the colonic tissue was hyperemia and edema,the colon became shorter and the intestinal wall became thicker,and inflammatory cells infiltrated the mucosa and submucosa.The mucosa was damaged and exfoliated.Erosion and ulcer could be seen.The gland structure of large intestine was destroyed.The infiltration of lymphocyte,plasma cell and monocyte could reach the muscular layer.In the positive control group and the high,middle and low dose groups of Ginseng Baidu Powder,the mucosal damage of colonic tissue was improved to varying degrees,congestion and edema were reduced,the structure and function of large intestinal glands were repaired,erosion repair and healing,ulcer area was reduced,inflammatory cell infiltration was reduced,granulation tissue appeared,etc.Colonic mucosal tissue injury score(TDI)showed that the positive control group had a significant improvement compared with the model group(P<0.05).(3)Gastrointestinal motility: Compared with model group,the gastric residual rate of high,middle and low dose groups and positive control group of Ginseng Baidu Powder decreased significantly(P<0.05).The small intestinal propulsive rate of the positive control group and the high,middle and low dose groups of Ginseng Baidu Powder increased(P<0.05).(4)Serum levels of IL-1beta,IL-6,TNF-alpha and IFN-gamma: Serum levels of IL-1beta,IL-6,TNF-alpha and IFN-gamma in model group increased significantly.Compared with the model group,the serum levels of IL-1beta and TNF-alpha in the positive control group and the high dose group of Ginseng Baidu Powder decreased(P<0.05).The serum levels of IL-6 and IFN-gamma in positive control group and high dose group of Ginseng Baidu Powder decreased(P<0.05).(5)Serum MDA,SOD and MPO levels: The content of MDA and MPO increased and the activity of SOD decreased in model group(P<0.05).Compared with the model group,MDA content in the positive control group and the high,middle and low dose groups of Ginseng Baidu Powder decreased(P<0.05),SOD activity in the middle dose group of Ginseng Baidu Powder increased(P<0.05),and MPO content in the positive control group decreased(P<0.05).(6)The expression of Occludin,Claudin-5 and ZO-1 in colon tissue: The expression of Occludin,Claudin-5 and ZO-1 in colon tissue of model group was significantly decreased(P<0.05).Compared with the model group,the expressions of Occludin,Claudin-5 and ZO-1 in the positive control group and the high,middle and low dose groups of Ginseng Baidu Powder increased significantly(P<0.05).(7)The expression levels of Occludin,Claudin-5 and ZO-1 protein in colon tissue: The expression levels of Occludin,Claudin-5 and ZO-1 in colon tissue of model group were significantly decreased(P<0.05).Compared with the model group,the expression of Occludin protein in colon tissue of the positive control group and the high,middle and low dose groups of Ginseng Baidu Powder increased significantly(P<0.05).The expression level of Claudin-5 protein in positive control group was significantly increased(P<0.05).The expression level of ZO-1 protein in positive control group and high and low dose groups of Ginseng Baidu Powder increased significantly(P<0.05).Conclusions: Intervention of "Ni Liu Wan zhou" theory on intestinal mucosa in rats with ulcerative colitis: It can significantly improve the general state of ulcerative colitis rats and the degree of colonic inflammation.It can promote the improvement of gastrointestinal motility function.It can inhibit the release of pro-inflammatory cytokines and improve the immune function of the body.It can reduce the content of oxygen free radicals and repair intestinal mucosal damage.It can regulate intestinal mucosal tight junction related proteins,up-regulate protein expression to maintain structural stability,reduce intestinal mucosal permeability,and protect intestinal mucosal mechanical barrier.