Protective Effect of Phillyrin on Lethal LPS-Induced Neutrophil Inflammation in Zebrafish
- Yang, L., Zhou, X., Huang, W., Fang, Q., Hu, J., Yu, L., Ma, N., Zhang, W.
- Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology 43: 2074-2087 (Journal)
- Registered Authors
- Ma, Ning, Zhang, Wenqing
- Inflammation, Lps, MyD88, NF-κB, Neutrophils, Phillyrin, Zebrafish
- MeSH Terms
- Inflammation/chemically induced*
- MAP Kinase Signaling System/drug effects
- NF-kappa B/metabolism
- Neutrophil Infiltration/drug effects
- Neutrophils/drug effects*
- Real-Time Polymerase Chain Reaction
- Tumor Necrosis Factor-alpha/metabolism
- 29059681 Full text @ Cell Physiol. Biochem.
Yang, L., Zhou, X., Huang, W., Fang, Q., Hu, J., Yu, L., Ma, N., Zhang, W. (2017) Protective Effect of Phillyrin on Lethal LPS-Induced Neutrophil Inflammation in Zebrafish. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology. 43:2074-2087.
Background/aims Forsythia suspensa Vahl. (Oleaceae) fruits are widely used in traditional Chinese medicine to treat pneumonia, typhoid, dysentery, ulcers and oedema. Antibacterial and anti-inflammatory activities have been reported for phillyrin (PHN), the main ingredient in Forsythia suspensa Vahl fruits, in vitro. However, the underlying mechanisms in vivo remain poorly defined. In this study, we discovered that PHN exerted potent anti-inflammatory effects in lethal LPS-induced neutrophil inflammation by suppressing the MyD88-dependent signalling pathway in zebrafish.
Methods LPS-yolk microinjection was used to induce a lethal LPS-infected zebrafish model. The effect of PHN on the survival of zebrafish challenged with lethal LPS was evaluated using survival analysis. The effect of PHN on neutrophil inflammation grading in vivo was assessed by tracking neutrophils with a transgenic line. The effects of PHN on neutrophil production and migration were analysed by SB+ cell counts during consecutive hours after modelling. Additionally, key cytokines and members of the MyD88 signalling pathway that are involved in inflammatory response were detected using quantitative RT-PCR. To assess gene expression changes during consecutive hours after modelling, the IL-1β, IL-6, TNF-α, MyD88, TRIF, ERK1/2, JNK, IκBa and NF-κB expression levels were measured.
Results PHN could protect zebrafish against a lethal LPS challenge in a dose-dependent manner, as indicated by decreased neutrophil infltration, reduced tissue necrosis and increased survival rates. Up-regulated IL-1β, IL-6 and TNF-α expression also showed the same tendencies of depression by PHN. Critically, PHN significantly inhibited the LPS-induced activation of MyD88, IκBa, and NF-κB but did not affect the expression of ERK1/2 MAPKs or JNK MAPKs in LPS-stimulated zebrafish. Additionally, PHN regulated the MyD88/IκBα/NF-κB signalling pathway by controlling IκBα, IL-1β, IL-6, and TNF-α expression.
Conclusion This study provides a rationale for the clinical application of PHN as an anti-inflammatory agent.
Genes / Markers
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes