Endotoxin Molecule Lipopolysaccharide-Induced Zebrafish Inflammation Model: A Novel Screening Method for Anti-Inflammatory Drugs

Molecules

Li-Ling Yang²  Guo-Quan Wang²  Li-Mei Yang¹  Zhi-Bing Huang³  Wen-Qing Zhang³ 

[1]School of Laboratory Medicine, Guangdong Medical College, Dongguan 523808, China

[2] E-Mail:

[3]School of Traditional Chinese Medicine, Southern Medical University, Guangzhou 510515, China

[4] E-Mails:

[5]The Key Laboratory of Zebrafish Modeling and Drug Screening for Human Diseases Institute, Southern Medical University, Guangzhou 510515, China

[6] E-Mail:

关键词: zebrafish;    LPS;    macrophages;    neutrophils;    screening;    qRT-PCR;    mortality;   

DOI  :  10.3390/molecules19022390

来源: mdpi

【 摘 要 】

Lipopolysaccharide (LPS), an endotoxin molecule, has been used to induce inflammatory responses. In this study, LPS was used to establish an in vivo inflammation model in zebrafish for drug screening. We present an experimental method that conveniently and rapidly assesses the anti-inflammatory properties of drugs. The yolks of 3-day post-fertilization (dpf) larvae were injected with 0.5 mg/mL LPS to induce fatal inflammation. After LPS stimulation, macrophages were tracked by NR and SB staining and neutrophil migration was observed using the MPO:GFP line. Larval mortality was used as the primary end-point. Expression levels of key cytokines involved in the inflammatory response including IL-1β, IL-6, and TNF-α, were measured using quantitative reverse transcription polymerase chain reaction (RT-PCR). Macrophages and neutrophils were both recruited to the LPS-injected site during the inflammatory response. Mortality was increased by LPS in a dose-dependent manner within 48 h. Analyses of IL-1β, IL-6, and TNF-α expression levels revealed the upregulation of the inflammatory response in the LPS-injected larvae. Further, the anti-inflammatory activity of chlorogenic acid (CA) was evaluated in this zebrafish model to screen for anti-inflammatory drugs. A preliminary result showed that CA revealed a similar effect as the corticosteroid dexamethasone (DEX), which was used as a positive control, by inhibiting macrophage and neutrophil recruitment to the LPS site and improving survival. Our results suggest that this zebrafish screening model could be applied to study inflammation-mediated diseases. Moreover, the Traditional Chinese Medicine CA displays potential anti-inflammatory activity.

【 授权许可】

CC BY   
© 2014 by the authors; licensee MDPI, Basel, Switzerland.