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Molecules 2014, 19(2), 2390-2409; doi:10.3390/molecules19022390

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

3,*  and 1,*
1 School of Traditional Chinese Medicine, Southern Medical University, Guangzhou 510515, China 2 School of Laboratory Medicine, Guangdong Medical College, Dongguan 523808, China 3 The Key Laboratory of Zebrafish Modeling and Drug Screening for Human Diseases Institute, Southern Medical University, Guangzhou 510515, China These authors contributed equally to this work.
* Authors to whom correspondence should be addressed.
Received: 11 January 2014 / Revised: 7 February 2014 / Accepted: 10 February 2014 / Published: 21 February 2014
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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.
Keywords: zebrafish; LPS; macrophages; neutrophils; screening; qRT-PCR; mortality zebrafish; LPS; macrophages; neutrophils; screening; qRT-PCR; mortality
This is an open access article distributed under the Creative Commons Attribution License (CC BY) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Yang, L.-L.; Wang, G.-Q.; Yang, L.-M.; Huang, Z.-B.; Zhang, W.-Q.; Yu, L.-Z. Endotoxin Molecule Lipopolysaccharide-Induced Zebrafish Inflammation Model: A Novel Screening Method for Anti-Inflammatory Drugs. Molecules 2014, 19, 2390-2409.

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