In Vivo High-throughput Screening Inhibition Of Macrophage Migration In Zebrafish Chinese Medicine

Objective:Establishment of green fluorescent protein (EGFP) marker of macrophages and macrophage migration and accumulation of transgenic zebrafish model (Zlyz-EGFP)1, and apply the model selection inhibition of zebrafish macrophages to acute injury site migration of Chinese medicine.Methods:Molecular cloning, microinjection, live video and other methods, the establishment of the green fluorescent protein as a marker of macrophages in transgenic zebrafish (Zlyz-EGFP), with a sharp sterile blades near the Ministry of the zebra fish at the cutting cycle, resulting in acute injury to the injury of the tail of macrophage migration and accumulation of acute inflammatory disease model, and gathered under the fluorescent microscope, the Ministry of zebra fish to count the number of macrophages. Application Zlyz-EGFP in acute inflammatory disease models in zebrafish screening inhibition of macrophage migration in zebrafish, the medicine together, and with the in vitro migration (transwell) system to verify the results.Results:We screened 296 Chinese herbs and found that 3#,28#,35 # medicine alcohol extract can significantly inhibit Zlyz-EGFP zebrafish macrophages to acute injury site migration, aggregation; also found that 3# of the ethanol extract material with 28# or 35# ethanol extract inhibited Zlyz-EGFP between the zebrafish macrophage migration to sites of acute injury there a synergistic effect; in vitro found that 3#,28#,35# medicine ethanol extract can significantly inhibit THP-1 cell chemotaxis and migration.Conclusion:1. We build Zlyz-EGFP by zebrafish and apply a sterile scalpel sharp cutting zlyz-EGFP was successfully zebra fish establishment of the Ministry of the ideal model of inflammatory disease;2.3#,28#,35# medicine can significantly inhibition Zlyz-EGFP zebrafish macrophages migrate to the site of acute injury, aggregation;3.3#,28# and 35# medicine on Zlyz-EGFP zebrafish macrophage migration to sites of acute injury, the effectiveness of a dose-dependent inhibition of aggregation;4.3# medicine in the 50mg/L with the 28#,35# medicine at 50mg /L when combined with inhibition of Zlyz-EGFP in zebrafish macrophage migration to sites of acute injury, there is a synergistic effect on the aggregation; 53#,28# and 35# medicine to sieve out will drive inflammation, autoimmune diseases and cancer and other diseases for basic research and clinical treatment.