Dasatinib regulates LPS-induced microglial and astrocytic neuroinflammatory responses by inhibiting AKT/STAT3 signaling
Dasatinib regulates LPS-induced microglial and astrocytic neuroinflammatory responses by inhibiting AKT/STAT3 signaling
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DOI：
10.1186/s12974-019-1561-x
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发表时间：
2019-10-26
影响因子：
9.3
通讯作者：
Hoe, Hyang-Sook
中科院分区：
文献类型：
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作者：
Ryu, Ka-Young;Lee, Hyun-Ju;Hoe, Hyang-Sook
fda批准的小分子药物达沙替尼目前用于治疗慢性髓性白血病（CML）。然而，达沙替尼对小胶质细胞和/或星形细胞神经炎症反应的影响及其作用机制尚未得到详细研究。方法用达沙替尼（100或250 nM）或载药（1% DMSO）处理BV2小胶质细胞、原代星形胶质细胞或原代小胶质细胞30 min或2 h，然后用脂多糖（LPS； 200 ng/ml或1 μ g/ml）或PBS处理5.5 h。免疫细胞化学;亚细胞分离;随后进行免疫组化，以确定达沙替尼对lps诱导的神经炎症的影响。此外，野生型小鼠注射达沙替尼（20 mg/kg，每天腹腔注射（i.p） 4天或20 mg/kg，每天口服（p.o） 4天或2周）或载药（4% DMSO + 30%聚乙二醇(PEG) + 5% Tween 80），然后注射LPS （10 mg/kg, i.p）或PBS。然后进行免疫组化，ELISA检测血浆IL-6、IL-1 β和tnf - α水平。结果达沙替尼调节lps诱导的BV2小胶质细胞、原代小胶质细胞和原代星形胶质细胞的促炎细胞因子和抗炎细胞因子水平。在BV2小胶质细胞中，达沙替尼通过调节TLR4/AKT和/或TLR4/ERK信号传导调节lps诱导的促炎细胞因子水平。此外，腹腔注射和口服达沙替尼可抑制lps诱导的小胶质细胞/星形胶质细胞活化、促炎细胞因子水平（包括脑和血浆水平）和野生型小鼠脑中性粒细胞滚动。结论：达沙替尼可调节lps诱导的脑内小胶质细胞和星形胶质细胞活化、促炎细胞因子水平和中性粒细胞滚动。
Background The FDA-approved small-molecule drug dasatinib is currently used as a treatment for chronic myeloid leukemia (CML). However, the effects of dasatinib on microglial and/or astrocytic neuroinflammatory responses and its mechanism of action have not been studied in detail. Methods BV2 microglial cells, primary astrocytes, or primary microglial cells were treated with dasatinib (100 or 250 nM) or vehicle (1% DMSO) for 30 min or 2 h followed by lipopolysaccharide (LPS; 200 ng/ml or 1 mu g/ml) or PBS for 5.5 h. RT-PCR, real-time PCR; immunocytochemistry; subcellular fractionation; and immunohistochemistry were subsequently conducted to determine the effects of dasatinib on LPS-induced neuroinflammation. In addition, wild-type mice were injected with dasatinib (20 mg/kg, intraperitoneally (i.p.) daily for 4 days or 20 mg/kg, orally administered (p.o.) daily for 4 days or 2 weeks) or vehicle (4% DMSO + 30% polyethylene glycol (PEG) + 5% Tween 80), followed by injection with LPS (10 mg/kg, i.p.) or PBS. Then, immunohistochemistry was performed, and plasma IL-6, IL-1 beta, and TNF-alpha levels were analyzed by ELISA. Results Dasatinib regulates LPS-induced proinflammatory cytokine and anti-inflammatory cytokine levels in BV2 microglial cells, primary microglial cells, and primary astrocytes. In BV2 microglial cells, dasatinib regulates LPS-induced proinflammatory cytokine levels by regulating TLR4/AKT and/or TLR4/ERK signaling. In addition, intraperitoneal injection and oral administration of dasatinib suppress LPS-induced microglial/astrocyte activation, proinflammatory cytokine levels (including brain and plasma levels), and neutrophil rolling in the brains of wild-type mice. Conclusions Our results suggest that dasatinib modulates LPS-induced microglial and astrocytic activation, proinflammatory cytokine levels, and neutrophil rolling in the brain.