Analysis of amphotericin B-induced cell signaling with chemical inhibitors of signaling molecules
Analysis of amphotericin B-induced cell signaling with chemical inhibitors of signaling molecules
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DOI：
10.1111/j.1348-0421.2006.tb03792.x
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发表时间：
2006-01-01
影响因子：
2.6
通讯作者：
Nakayama, K
中科院分区：
文献类型：
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作者：
Matsuo, K;Hotokezaka, H;Nakayama, K
虽然阿替霉素B（AmB）是一种主要的抗侵袭性真菌感染的多烯抗生素，但对患者施用有时会引起炎症副作用，这限制了抗生素的使用。我们研究了AmB诱导的细胞内信号传导。核因子-κ B（NF-κ B B）的p65（RelA）被单核细胞样细胞系RAW 264.7中的AmB磷酸化。在信号分子的化学抑制剂中，U-73122（磷脂酶C（PLC）抑制剂）、Go 6976（蛋白激酶C（PKC）抑制剂）、BAPTA-AM（钙螯合剂）、LFM-A13（布鲁顿酪氨酸激酶（Btk）特异性抑制剂）和PP 2（c-Src激酶抑制剂）抑制AmB诱导的p65磷酸化和p65向细胞核的移位。U-73122和Go 6976减少了RAW 264.7细胞中AmB介导的促炎细胞因子（肿瘤坏死因子（TNF）-α和白细胞介素（IL）-6）的诱导。此外，AmB诱导的NF-κ B活化在toll样受体（TLR）2表达的细胞中观察到，并且NF-κ B活化被U-73122抑制，而肽聚糖诱导的NF-κ B活化（也依赖于TLR 2）不被U-73122抑制。最后，U-73122部分抑制BALB/c小鼠体内由AmB给药诱导的TNF-α和IL-6的产生。这些结果表明，从AmB刺激到促炎细胞因子产生的信号转导是由TLR 2、Btk、PLC、PKC、c-Src和NF-κ B介导的。这些信号分子可能成为化疗抑制AmB诱导的促炎细胞因子产生的靶点。
Although amphotericin B (AmB) is a major polyene antibiotic against invasive fungal infection, administration to patients sometimes causes inflammatory side effects, which limits the usage of the antibiotic. We studied the intracellular signaling that was induced by AmB. p65 (RelA) of nuclear factor-kappa B (NF-kappa B), a well-known signaling molecule as an inducer of proinflammatory cytokines, was phosphorylated by AmB in RAW264.7 cells, a monocyte-like cell line. Among chemical inhibitors of signaling molecules, U-73122 (phospholipase C (PLC) inhibitor), Go6976 (protein kinase C (PKC) inhibitor), BAPTA-AM (calcium chelator), LFM-A13 (Bruton's tyrosine kinase (Btk)-specific inhibitor), and PP2 (c-Src kinase inhibitor) suppressed AmB-induced phosphorylation of p65 and translocation of p65 into the nucleus. U-73122 and Go6976 reduced AmB-mediated induction of proinflammatory cytokines (tumor necrosis factor (TNF)-alpha and interleukin (IL)-6) in RAW264.7 cells. Furthermore, AmB-induced activation of NF-kappa B was observed in toll-like receptor (TLR) 2-expressed cells, and the activation of NF-kappa B was inhibited by U-73122, whereas peptidoglycan-induced NF-kappa B activation, which was also dependent on TLR2, was not inhibited by U-73122. Finally, U-73122 partially suppressed in vivo production of TNF-alpha and IL-6 induced by AmB administration in BALB/c mice. These results suggested that the signaling from AmB stimulation to proinflammatory cytokine production is mediated by TLR2, Btk, PLC, PKC, c-Src and NF-kappa B. These signaling molecules may become a target for chemotherapy suppressing AmB-induced proinflammatory cytokine production.