兔鱼(Siganus fuscesens)巨噬细胞样细胞系的表征:黄芩提取物抗炎反应的例证

Yi-Teng Xia , Edwin Hok-Chi Cheng , Ying-Jie Xia , Qi-Yun Wu , Lynn Hoi-Lam Zhang , Sheng-Ying Lin , Tina Ting-Xia Dong , Qi-Wei Qin , Wen-Xiong Wang , Karl Wah-Keung Tsim
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引用次数: 8

摘要

从兔鱼(Siganus fuscescens)头肾中分离到一株新的细胞系并进行了鉴定。新的巨噬细胞样细胞系被命名为兔鱼巨噬细胞(RFM),该细胞自发育以来可传代培养50多个周期。对RFM细胞株在不同温度和血清浓度下的生长进行了测试:在28℃下,20%血清的生长条件最佳。在培养的RFM细胞中,18S rRNA测序、细胞角蛋白和cd68免疫染色证实其为fuscescens巨噬细胞。培养的RFM细胞暴露于LPS触发的炎症刺激下,对炎症表现出高度敏感的反应,包括一氧化氮的释放、细胞因子的表达和吞噬的激活。黄芩地上部水提物SBA对fuscescens鱼具有抗炎作用。为了扩大SBA在水产养殖中的应用,将其提取物及其有效类黄酮黄芩苷和黄芩苷应用于lps处理的RFM细胞。应用SBA提取物、黄芩苷或黄芩苷可抑制lps诱导的炎症细胞因子IL-1β、TNF-α的表达以及转录因子NF-κB的信号传导。结果表明,所建立的RFM细胞系可作为炎症相关药物筛选的理想体外模型。此外,SBA草药提取物在水产养殖中的概念得到了其抗炎症功效的支持。
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Characterization of a macrophagic-like cell line derived from rabbit fish (Siganus fuscescens): An illustration of anti-inflammatory responses of the herbal extract of Scutellaria baicalensis

A new cell line was isolated and characterized from the head kidney of Siganus fuscescens (rabbit fish). The new macrophagic-like cell line was named as rabbit fish macrophage (RFM), and which could be sub-cultured for over 50 cycles since the development. RFM cell line was tested for growth in different temperatures and serum concentrations: the best growing condition was optimized at 20% serum under 28 °C. In cultured RFM cells, sequencing of 18S rRNA, as well as immunostaining of cytokeratin and CD 68, confirmed the identity as macrophagic cell of S. fuscescens. Cultured RFM cells were exposed to challenge of inflammation, as triggered by LPS, showing highly sensitive responses to inflammation, including release of nitric oxide, expression of cytokine, and activation of phagocytosis. The water extract of aerial part of Scutellaria baicalensis, named as SBA, has been shown anti-inflammatory property in S. fuscescens fish. In order to extend the application of SBA in aquaculture, the extract and its effective flavonoids, i.e. baicalin and scutellarin, were applied in LPS-treated RFM cells. Application of SBA extract, baicalin or scutellarin, inhibited the expressions of LPS-induced inflammatory cytokines, i.e. IL-1β, TNF-α, as well as the signaling of transcription factor NF-κB. The results support the established RFM cell line could be an ideal in vitro model in drug screening relating to inflammation. Additionally, the notion of SBA herbal extract in fish aquaculture is supported by its efficacy against inflammation.

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