Effect Of Anchang Decoction On TLR4,p38 MAPK And IL-17 In Colon Tissue Of IBS-D Rats Based On Inflammatory Mechanism

Objective: In this study,pepsin gavage combined with binding and tail clip stress was used to prepare Irritable Bowel Syndrome with diarrhea(IBS-D)model rat to observe the changes of colon histopathology,TLR4,p38 MAPK,IL-17 protein and m RNA of Anchang decoction on the colon of IBS-D model rat,and to clarify that Anchang decoction can improve visceral hypersensitivity by inhibiting the expression of inflammatory factors.Relieve IBS-D abdominal pain,diarrhea and other symptoms,and provide relevant theoretical basis for clinical diagnosis and treatment of diarrhea-type irritable bowel syndrome.Methods: Forty clean male SD rats were randomly divided into normal group,model group,western medicine group and Anchang group with 10 rats in each group.Except for normal group,model group,western medicine group and Anchang decoction group were treated with capsaicin gavage combined with binding and tail-clip stress to replicate IBS-D model for 2 weeks.After the successful evaluation of IBS-D model,the western medicine group was given15mg/kg Pivillium bromide tablet intragastric administration,while the Anchang decoction group was given Anchang decoction 6.2g/kg intragastric administration,once a day for 2 weeks.During the experiment,the general condition,fecal character,body weight change and AWR score of rats in each group were closely observed.After the drug intervention,all rats were killed and colonic tissue specimens were collected,hematoxylin eosinstaining(HE)was applied to compare the colonic histopathology of different groups of rats.Western Blot was used to detect the expression levels of TLR4,p38 MAPK,p-p38 MAPK and IL-17 protein in colon tissues of rats in each group,and RT-PCR was used to detect the m RNA expression levels of TLR4,p38 MAPK and IL-17 in colon tissues of rats in different groups.Results:(1)The general situation of rats: the rats in the normal group had good mental state,quick movement,normal eating,drinking and stool,white and glossy hair;After the successful preparation of IBS-D model,the rats in model group,western medicine group and Anchang decoction group showed listless spirit,dull response,decreased food intake and activity,decreased body weight,increased stool frequency,fecal tissue was rotten and unformed,and the hair of the rats became yellow,dry and unglossy,and even fell off.After drug intervention,the mental state,appetite,body weight,range of motion,hair and stool of rats were improved.(2)Changes in body weight of rats: Before modeling,there was no significant difference in body weight of rats among all groups(P>0.05);After modeling,the body weight of rats in model group,western medicine group and Anchang decoction group was significantly reduced compared with that in normal group(P< 0.01).After drug intervention,the body weight of rats in western medicine group and Anchang decoction group was significantly increased compared with model group(P< 0.01);Compared with western medicine group,the weight increase of rats in Anchang decoction group was higher(P< 0.05).(3)Rat abdominal withdrawal reflex(AWR): Under different balloon pressure levels,the AWR score of model group was significantly increased compared with normal group(P<0.01).After drug intervention,AWR score of rats in western medicine group and Anchang decoction group was decreased compared with model group(P< 0.05).(4)HE staining of colon tissue of rats in each group:no abnormal pathological changes were observed in colonic mucosa of rats in the normal group under microscope.In the model group,only a small number of inflammatory cells were locally infiltrated in the colonic mucosa.After drug treatment,the inflammatory infiltration of colonic mucosa was reduced in the western medicine group,but no inflammatory cells were found in the colonic mucosa of the rats in the Anchang decoction group.(5)Expression levels of TLR4,p38 MAPK,p-p38 MAPK and IL-17 in rat colon tissue:Compared with normal group,the expression levels of TLR4,p-p38 MAPK and IL-17 protein in colon tissues of rats in model group were increased(P<0.01),and the expression of p38 MAPK protein was also increased(P<0.05).After drug treatment,the protein expression levels of TLR4,p-p38 MAPK and IL-17 in colon tissues of rats in western medicine group and Anchang decoction group were significantly decreased compared with model group(P<0.01).The protein expression levels of TLR4,p-p38 MAPK and IL-17 in colon tissue of rats in Anchang decoction group were lower than those in western medicine group(P<0.05).(6)m RNA expression levels of TLR4,p38 MAPK and IL-17 in colon tissues of rats: Compared with normal group,m RNA expression levels of TLR4,p38 MAPK and IL-17 in colon tissues of rats in model group were significantly up-regulated(P<0.01);After drug treatment,compared with model group,m RNA expression levels of TLR4,p38 MAPK and IL-17 in colon tissues of rats in western medicine group and Anchang decoction group were significantly decreased(P<0.01);The m RNA expression levels of TLR4,p38 MAPK and IL-17 in Anchang decoction group were lower than those in western medicine group(P< 0.05).Conclusions:(1)Anchang decoction can improve high visceral sensitivity and reduce intestinal inflammation in IBS-D rats.(2)Anchang decoction can inhibit the activation of TLR4 and p38 MAPK signaling pathways,reduce the release of inflammatory factors,improve intestinal inflammatory response and reduce visceral hypersensitivity.