Benvitimod Is A Potential Drug Of Inflammatory Bowel Disease Through Protecting Gut Barrier And Promoting Apoptosis Of Pro-inflammation Immune Cells

Background:Inflammatory bowel disease is a chronic and non-specific intestinal inflammatory disease,typical of immune related.Its pathogenesis is manifested by abnormal immune responses mediated by commensal bacteria in genetically susceptible populations.Existing drugs are difficult to maintain long-term remission of the disease,and the process of repeated medication promotes drug tolerance of patients.Therefore,it is crucial to explore drugs which share different mechanism from existing products.Benvitimod is a new type of non-steroidal small molecule drug.As an agonist of aryl hydrocarbon receptors,it can regulate immunity and inhibit the expression of various inflammatory factors.Aromatic hydrocarbon receptors are widely distributed and can receive signals from inflammatory factors,some toxic compounds in the environment and endogenous aromatic hydrocarbon ligands,and are activated to promote the expression of downstream genes,thereby causing corresponding biological effects.Its effect is determined by its basal expression level and the ligand content of aryl hydrocarbon receptors in the environment,and its activation mechanism is complex,which is not directly related to the expression content of the protein.Aryl hydrocarbon receptors can bind to ubiquitin proteins for degradation.However,whether the ubiquitination modification of aryl hydrocarbon receptors is the mechanism by which benvitimod achieves its pharmacological effects remains to be further studied.Aims:This research is to verify the effectiveness of benvitimod to IBD in vitro and in vivo.Methods:In this study,through in vivo experiments,mice were given an enema to demonstrate the therapeutic effect of benvitimod by measuring body weight,disease activity index,and colon length.In the in vitro model,according to the expression level of AhR,two cell lines,NCM460 and LoVo,were screened,and H9 cells were added to test the proliferation and apoptosis level of the cells after stimulation with benvitimod.The relationship between the effect of benvitimod and the level of AhR.Further,through the barrier test and the detection of tight junction protein,the effect of benvitimod and AhR on the barrier was proved.On the other hand,the effect of benvitimod and AhR on immunity was detected by detecting the lamina propria lymphoid of gene knockout mice(VillinCre/CD4Cre).The content of cell CD4+AhR+cell population,the effect of benvitimod on the apoptosis level of lamina propria lymphocytes in knockout mice was realized.Finally,the molecular mechanism of the effect of benvitimod was partially explored by detecting the downstream pathway indicators and studying the ubiquitination level of AhR.Results:1.The enema of the benvitimod with sodium hydrogen added enhanced therapeutic effect of IBD,including improving systematic symptoms and alleviating the colon carcinogenesis;2.Benvitimod maintained the gut barrier by promoting normal epithelial proliferation and inhibited growth of cancer cells;3.Benvitimod inhibit immunity via AhR;4.AhR increased the sensitivity of tissue to the inflammatory factors,and lower level of AhR revealed relieve of inflammation;5.Benvitmod decreased AhR ubiquitination and downregulated the HIF-1?.Conclusions:The novel benvitimod enema is effective in treating inflammatory bowel disease,with lower safety risk.It maintains the AhR-dependent gut barrier and promotes apoptosis of pro-inflammation cells.Benvitimod is estimated to be a potential drug in IBD.