Primary Study Of The Effects Of Ibuprofen On Mycobacterium Tuberculosis Infection Using Zebrafish Model

As an excellent model to study innate immunity of vertebrates,zebrafish provides us with the possibility of imaging host-pathogen interaction in vivo.Lipopolysaccharide(LPS)is an endotoxin molecule,which has been used to induce inflammatory reaction.In this study,the inflammation model of zebrafish was established by LPS,which was used to detect the anti-inflammatory properties of ibuprofen.Immerse the 2 days postfertilization(dpf)larvae with 10ug/ml LPS for 24 h to induce inflammatory reaction.Quantitative reverse transcription polymerase chain reaction(RT-q PCR)was used to detect cytokines related to inflammatory reaction.The expression levels of mmp9,mmp13 a,irg1l,il6 st,lyz,mpeg1.2 were analyzed,and the inflammatory response of LPS-immersion larvae was up-regulated.Moreover,ibuprofen has been evaluated to have certain influence on pro-inflammatory factors and anti-inflammatory factors.In addition,zebrafish inflammation was established by caudal fin excision,neutrophils and macrophages were tracked by Sudan black B and neutral red staining respectively,and myeloid cells and macrophages were tracked by using transgenic zebrafish lines cz13 and mfap4 respectively.Our results show that ibuprofen can inhibit the recruitment of macrophages and neutrophils to inflammatory sites.We also studied zebrafish-M.marine infection model to improve our understanding of innate immune system.We analyzed the transcriptome of pattern recognition receptors and other cytokines in zebrafish infected with M.marine.The results showed that the m RNA levels of pattern recognition receptors(PRRs)and cytokines generally increased after pathogen invasion.The pretreatment of ibuprofen could affect the m RNA levels of cytokines,but it had no significant effect on PRRs.This study systematically studied the specific regulatory genes of ibuprofen in the inflammatory response,and found a number of new inflammatory response and innate immune response genes affected by ibuprofen,providing a preliminary research basis for studying the possible unknown side effect mechanism of ibuprofen.