title: Meisoindigo, but not its core chemical structure indirubin, inhibits zebrafish interstitial leukocyte chemotactic migration.
abstract: BACKGROUND
  Inflammatory disease is a big threat to human health. Leukocyte chemotactic migration is required for efficient inflammatory response. Inhibition of leukocyte chemotactic migration to the inflammatory site has been shown to provide therapeutic targets for treating inflammatory diseases.
  
  
  OBJECTIVE
  Our study was designed to discover effective and safe compounds that can inhibit leukocyte chemotactic migration, thus providing possible novel therapeutic strategy for treating inflammatory diseases.
  
  
  METHODS
  In this study, we used transgenic zebrafish model (Tg:zlyz-EGFP line) to visualize the process of leukocyte chemotactic migration. Then, we used this model to screen the hit compound and evaluate its biological activity on leukocyte chemotactic migration. Furthermore, western blot analysis was performed to evaluate the effect of the hit compound on the AKT or ERK-mediated pathway, which plays an important role in leukocyte chemotactic migration.
  
  
  RESULTS
  In this study, using zebrafish-based chemical screening, we identified that the hit compound meisoindigo (25 μM, 50 μM, 75 μM) can significantly inhibit zebrafish leukocyte chemotactic migration in a dose-dependent manner (p = 0.01, p = 0.0006, p < 0.0001). Also, we found that meisoindigo did not affect the process of leukocyte reverse migration (p = 0.43). Furthermore, our results unexpectedly showed that indirubin, the core structure of meisoindigo, had no significant effect on zebrafish leukocyte chemotactic migration (p = 0.6001). Additionally, our results revealed that meisoindigo exerts no effect on the Akt or Erk-mediated signalling pathway.
  
  
  CONCLUSIONS
  Our results suggest that meisoindigo, but not indirubin, is effective for inhibiting leukocyte chemotactic migration, thus providing a potential therapeutic agent for treating inflammatory diseases.
authors.affiliation: a Department of Hematology , Renmin Hospital of Wuhan University , Wuhan , Hubei , China.|b State Key Laboratory of Medical Genomics and Shanghai Institute of Hematology , Rui Jin Hospital, Shanghai Jiao Tong University School of Medicine , Shanghai , China.
authors.forename: Baixin
authors.initials: B
authors.lastname: Ye
authors.affiliation: c Department of Neurosurgery , Renmin Hospital of Wuhan University , Wuhan , Hubei , China.
authors.forename: Xiaoxing
authors.initials: X
authors.lastname: Xiong
authors.affiliation: d College of Chemistry and Molecular Science , Wuhan University , Wuhan , Hubei , China.
authors.forename: Xu
authors.initials: X
authors.lastname: Deng
authors.affiliation: e Central Laboratory , Renmin Hospital of Wuhan University , Wuhan , Hubei , China.
authors.forename: Lijuan
authors.initials: L
authors.lastname: Gu
authors.affiliation: a Department of Hematology , Renmin Hospital of Wuhan University , Wuhan , Hubei , China.
authors.forename: Qiongyu
authors.initials: Q
authors.lastname: Wang
authors.affiliation: f Department of Pathology , Renmin Hospital of Wuhan University , Wuhan , Hubei , China.
authors.forename: Zhi
authors.initials: Z
authors.lastname: Zeng
authors.affiliation: e Central Laboratory , Renmin Hospital of Wuhan University , Wuhan , Hubei , China.
authors.forename: Xiang
authors.initials: X
authors.lastname: Gao
authors.affiliation: a Department of Hematology , Renmin Hospital of Wuhan University , Wuhan , Hubei , China.
authors.forename: Qingping
authors.initials: Q
authors.lastname: Gao
authors.affiliation: b State Key Laboratory of Medical Genomics and Shanghai Institute of Hematology , Rui Jin Hospital, Shanghai Jiao Tong University School of Medicine , Shanghai , China.
authors.forename: Yueying
authors.initials: Y
authors.lastname: Wang
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D000893:Anti-Inflammatory Agents / Q000737:chemistry / Q000494:pharmacology*; D002634:Chemotaxis, Leukocyte / Q000187:drug effects*; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D007211:Indoles / Q000737:chemistry / Q000494:pharmacology; D007249:Inflammation / Q000276:immunology / Q000378:metabolism / Q000517:prevention & control*; D007962:Leukocytes / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D015394:Molecular Structure; D015398:Signal Transduction / Q000187:drug effects; D013329:Structure-Activity Relationship; D013997:Time Factors; D015027:Zebrafish / Q000196:embryology / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
chemical_list: D000893:Anti-Inflammatory Agents; D007211:Indoles; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; C047330:N-methylisoindigotin; C027185:indirubin
keywords: Meisoindigo; indirubin; inflammation; leukocyte chemotactic migration; zebrafish