| Objective: to observe the effect of Ginseng Baidu Powder on AMPK/ULK1 autophagy signal pathway and intestinal mucosal barrier tight junction protein in rats with ulcerative colitis,and to explore the mechanism of this prescription in the treatment of ulcerative colitis.Methods: in this study,the rat model of ulcerative colitis was established by TNBS/-ethanol compound method,and the rats were randomly divided into blank group,model group,positive control group,high,middle and low dose Ginseng Baidu San groups(hereinafter referred to as Baidusan-H,M,L groups).(1)histopathological changes of rat colon were observed with naked eye and light microscope.(2)the expression of AMPKα m RNA in colon tissue was detected by real-time quantitative polymerase chain reaction.(3)The expressions of tight junction protein claudin-2,occludin,autophagy marker proteins LC3 II and P62,and AMPK/ULK1 pathway related proteins p-AMPK and p-ULK1 were detected by Western blotting.Results:(1)the changes of colonic tissue were observed with naked eye: in the model group,the colonic mucosa was obviously edema and hyperemia,ulcer adhesion was obvious,and the colon length was shortened;the edema and hyperemia ulcer in each treatment group was significantly alleviated,and the colon length was increased.(2)observation of pathological changes of colon tissue under microscope: in the model group,there were large area ulcer in colon mucosa,hyperplasia of connective tissue in submucosa and severe edema,accompanied by a large number of inflammatory cell infiltration,severe dilatation of intestinal gland,and(TDI)increased significantly(P<0.05).Compared with the model group,edema and inflammatory cell infiltration could still be seen in the local submucosa of each treatment group,but it was significantly less than that in the model group.TDI in the positive group and Baidu powder-M group decreased significantly(P<0.05).(3)content of tight junction protein occludin and claudin-2 in colon tissue: compared with the blank group,the expression level of occludin protein in the model group was significantly lower and the expression level of claudin-2 protein was significantly higher than that in the model group,while the expression of occludin in the Baidu San-M group was significantly higher than that in the model group(P< 0.05),and the expression level of claudin-2protein in each treatment group was significantly decreased(P<0.05).(4)content of autophagy-related protein P62 and LC3BII/I in colon tissue: compared with the blank group,the expression of P62 protein in the model group was significantly increased and LC3BII/I was significantly decreased,compared with the model group,the expression of P62 in each treatment group was significantly lower than that in the model group,and the LC3BII/I in each treatment group was significantly increased(P<0.05).(5)AMPK α m RNA expression in colon tissue: compared with the blank group,the AMPK α m RNA level in the model group was significantly decreased,and that in each treatment group was significantly higher than that in the model group(P<0.05).(6)expression of p-AMPK and p-ULK1 in colon tissue: the expression of p-AMPK and p-ULK1 in model group was significantly decreased,while the expression of p-AMPK in positive group,Baidu powder-M group and Baidu San-L group was significantly increased,and the expression of p-ULK1 in each treatment group was significantly increased(P<0.05).Conclusion:(1)Ginseng Baidu Powder can improve UC to a certain extent: it can improve colonic mucosal tissue injury,reduce inflammatory cell infiltration and repair intestinal mucosal barrier function in UC rats.(2)one of the mechanisms of Ginseng Baidu Powder in improving UC: it may activate autophagy through AMPK/ULK1 autophagy signal pathway,thus increase the transformation of LC3 I to LC3 II,degrade P62,reduce claudin-2,and increase the expression level of occludin in colon tissue,thus repair intestinal epithelial tight junction,reduce intestinal mucosal permeability,reduce inflammatory reaction,protect intestinal mucosal mechanical barrier and play a role in the treatment of ulcerative colitis. |
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