Pharmacological manipulation of inflammation resolution during spontaneously resolving tissue neutrophilia in the zebrafish
Pharmacological manipulation of inflammation resolution during spontaneously resolving tissue neutrophilia in the zebrafish
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DOI：
10.1189/jlb.0409255
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发表时间：
2010-02-01
影响因子：
5.5
通讯作者：
Renshaw, Stephen A.
中科院分区：
文献类型：
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作者：
Loynes, Catherine A.;Martin, Jane S.;Renshaw, Stephen A.
斑马鱼是对炎症等生理过程进行药理学操作的独特模型；它们很小，对许多小分子化合物具有渗透性，而且是透明的，可以通过观察转基因标记的炎症细胞群来可视化和定量炎症反应。利用体内特异性标记中性粒细胞的转基因品系(mpx：GFP)，我们研究了一系列药物对体内炎症消退的影响。这些药物的选择是因为它们在体外调节人中性粒细胞的功能和寿命。延缓中性粒细胞凋亡的药物(内毒素、DBcAMP和几种半胱氨酸氨基转移酶抑制剂)都会导致中性粒细胞炎症的消退延迟。反过来，绿青素和玫瑰花碱(中性粒细胞凋亡的诱导剂)导致中性粒细胞数量减少。通过时间推移分析观察细胞凋亡的发生，并通过中性粒细胞特异性mpx活性的双重染色(TSA染色)和凋亡标记物(TUNEL)的双重染色来确认细胞的发生。在炎症过程中，巨噬细胞跟随中性粒细胞进入炎症部位，巨噬细胞内可见TUNEL/TSA双阳性物质，这与其摄取凋亡的中性粒细胞一致。与哺乳动物模型相比，该模型有几个优点，并有助于研究药物调节炎症。J.Leukoc。比奥尔。87：203-212；2010。
Zebrafish are a unique model for pharmacological manipulation of physiological processes such as inflammation; they are small and permeable to many small molecular compounds, and being transparent, they permit the visualization and quantitation of the inflammatory response by observation of transgenically labeled inflammatory cell populations. Using a transgenic line specifically labeling neutrophils in vivo (mpx:GFP), we studied the effects of a range of pharmacological agents on the resolution of inflammation in vivo. These agents were selected for their ability to modulate neutrophil function and lifespan in human neutrophils in vitro. Agents delaying neutrophil apoptosis (LPS, dbcAMP, and several caspase inhibitors) all lead to a delay in resolution of neutrophilic inflammation. Reciprocally, pyocyanin and roscovitine (inducers of neutrophil apoptosis) lead to reduced neutrophil numbers. The occurrence of apoptosis was observed by time-lapse analysis and confirmed by dual staining for neutrophil-specific mpx activity (TSA staining) and an apoptotic marker (TUNEL). During inflammation, macrophages follow neutrophils into the inflamed site, and TUNEL/TSA dual-positive material can be demonstrated within macrophages, consistent with their uptake of apoptotic neutrophils. This model has several advantages over mammalian models and lends itself to the study of pharmaceutical agents modulating inflammation. J. Leukoc. Biol. 87: 203-212; 2010.