Study On The Therapeutic Effect And Mechanism Of Yujin Powder On Dampness-Heat Diarrhea In Mice

Dampness-heat diarrhea is a common disease in Chinese veterinarian clinic.Dampness-heat diarrhea is caused by dampness and heat in the intestines and mainly diarrhea.Dampness and heat discharge is a common disease that has long troubled breeding farms and pig farmers.Antimicrobial drugs were used in the treatment.Although antibiotics play a certain role in the treatment of this disease,drug resistance is also becoming more and more serious.Traditional Chinese medicine is not easy to remain in animal products,less toxic and side effects,improve animal resistance and other advantages have attracted much attention.Yujin powder,a traditional Chinese medicine compound,is a famous prescription for the treatment of dampness-heat diarrhea in the book "Yuan heng liao ma ji".It is composed of 8 medicines,including Yujin powder,rhizoma coptis,scutellaria,phyllocypress,gardenia fruit,white peony root,chebula chebula and rhubarb.It has the effect of clearing away heat and detoxification,astringent bowel and stopping diarrhea,and bacteria are not easy to develop drug resistance.In this study,BALB /c mice were used as research subjects to evaluate the therapeutic effect of traditional Chinese medicine Yujin Powder on dampness and heat diarrhea in mice,and to explore its effects on intestinal microorganism and TLR4/My D88/NF-k Bp65 signaling pathway.The purpose is to provide the basis for the rational application of this drug in clinic.Firstly,the dampness-heat dissipation model of mice was established by using high sugar and high fat,hunger disorder,high temperature and humidity,drinking alcohol and intraperitoneal injection of E.coli to simulate the disease environment.After successful replication,Yujin powder was used for treatment and gentamicin was used as control.Clinical symptoms,feed intake,water intake,body temperature and body weight of the experimental animals were observed and recorded daily.The pathological changes of intestinal tissue were observed by H.E.staining and transmission electron microscopy.Illumina Mi Seq technology was used to perform high-throughput sequencing of the feces of mice from different treatment groups,and species composition,Alpha diversity,principal coordinate analysis and structure were analyzed.The gene expressions of IL-6,IL-8,TNF,VEGF,P53,Fos,Casp3,TLR4,My D88 and NF-κBp65 were detected by Real-time PCR.TLR4,My D88 and NF-κBp65protein expressions were detected by Western-blot.The therapeutic effect of Yujin powder on dampness and heat relief was comprehensively evaluated from different levels,such as whole body,tissue,cell and molecule,and the network pharmacological method was used to screen the target of Yujin powder.The results showed that the model mice with dampness-heat diarrhea had disorderly hair,loss of weight and mental tiredness.Loose stool or watery stool,yellowish brown stool,foul-smelling stool,loose stool around the anus and tail;Many viscera hemorrhage,degeneration,necrosis.Compared with the normal control group,there were more lymphocytes and goblet cells in the model group,indicating chronic inflammation.At the same time,there were denatured necrosis and exfoliation of epithelial cells.Transmission electron microscopy showed that elliptical mitochondria in cells were observed in the normal control group,while obvious mitochondrial injury and rupture of the inner mitochondrial ridge were observed in the colon of the model group.After treatment with Yujin Powder,the histomathological changes mentioned above were significantly alleviated.The effect of Qingda control group was not as obvious as that of Yujin Powder.The pathological changes in the untreated group were more serious.The possible targets of Yujin powder were screened by network pharmacology method,and it was found that the core targets of Yujin powder treatment might be IL-6,IL-8,TNF,VEGF,P53,FOS,Casp3,etc.,mainly involved in TNF signaling pathway,toll-like receptor signaling pathway and cancer pathway.Real-time PCR results showed that Yujin Powder could significantly reduce the m RNA expression levels of IL-6,IL-8,TNF,VEGF,P53,FOS,Casp3 and the protein expression levels of TLR4/My D88 /NF-κBp65.It was proved that TLR4/ My D88 /NF-κBp65 signal transduction pathway was activated in the colon tissue of damp-heat relieving mice,and Yujin Powder could reduce the intestinal inflammatory response and intestinal mucosal injury by affecting the expression of the above targets.This study further confirmed the therapeutic effect and target of Yujin Powder on dampness-heat diarrhea syndrome,and also provided more options for clinical treatment of dampness-heat diarrhea syndrome.