Atsttrin reduces lipopolysaccharide-induced neuroinflammation by inhibiting the nuclear factor kappa B signaling pathway
Atsttrin reduces lipopolysaccharide-induced neuroinflammation by inhibiting the nuclear factor kappa B signaling pathway
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DOI：
10.4103/1673-5374.259623
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发表时间：
2019-11
影响因子：
6.1
通讯作者：
Lian Liu;Yuan Qu;Yi Liu;Hua Zhao;He Ma;A. Noor;Chang-Jiao Ji;L. Nie;M. Si;Lei Cheng
中科院分区：
文献类型：
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作者：
Lian Liu;Yuan Qu;Yi Liu;Hua Zhao;He Ma;A. Noor;Chang-Jiao Ji;L. Nie;M. Si;Lei Cheng
在神经炎性小鼠模型中，原颗粒与神经元存活密切相关，并可减轻炎症反应。Atsttrin是一种由三个原颗粒片段组成的工程蛋白质，已被证明具有与原颗粒相似的作用。艾特林在多种关节炎小鼠模型中具有抗炎作用，并可防止关节炎的进一步发展。然而，Atsttrin是否在神经炎症中起作用仍有待阐明。本研究通过侧脑室注射1μL脂多糖(10μg/μL)复制神经炎性小鼠模型。在侧脑室注射1μL脂多糖(10μg/μL)前，每隔3d腹腔注射艾司特林2.5 mg/kg，连续7d。另外，用0、100、300 ng/mL脂多糖处理星形胶质细胞，同时加入200 ng/mLAtsttrin。免疫组织化学、酶联免疫吸附试验和实时定量逆转录-聚合酶链式反应检测炎症介质的蛋白和mRNA水平，评估核因子-kappaB信号通路的激活。给予脂多糖后，野生型小鼠大脑和星形胶质细胞培养物中原颗粒蛋白的表达增加。给予脂多糖后，前颗粒蛋白基因敲除小鼠脑内肿瘤坏死因子-α、白介素1β和诱导型一氧化氮合酶的蛋白和基因表达水平均升高。艾特林治疗可降低脂多糖诱导的前颗粒蛋白基因敲除小鼠脑内肿瘤坏死因子-α、白介素1β、基质金属蛋白酶-3和诱导型一氧化氮合酶的蛋白和mR NA水平。艾特林还能降低脂多糖诱导的星形胶质细胞环氧合酶-2、诱导型一氧化氮合酶和基质金属蛋白酶3mRNA的表达，降低培养上清液中肿瘤坏死因子-α和白介素1β的浓度。此外，爱特灵显著降低脂多糖基因敲除小鼠和星形胶质细胞中磷酸化核因子kappa B抑制物a的水平，并降低星形胶质细胞中核因子kappaB的表达。综上所述，我们的研究结果表明，艾特林的抗神经炎作用与抑制核因子-kappaB信号通路有关，提示艾特林可能在神经炎症治疗中具有临床应用潜力。本研究由山东大学齐鲁医院动物伦理委员会中国(批准号：Kyll-2015(KS)-088)2015年2月10日。
Progranulin is closely related to neuronal survival in a neuroinflammatory mouse model and attenuates inflammatory reactions. Atsttrin is an engineered protein composed of three progranulin fragments and has been shown to have an effect similar to that of progranulin. Atsttrin has anti-inflammatory actions in multiple arthritis mouse models, and it protects against further arthritis development. However, whether Atsttrin has a role in neuroinflammation remains to be elucidated. In this study, we produced a neuroinflammatory mouse model by intracerebroventricular injection of 1 μL lipopolysaccharide (10 μg/μL). Atsttrin (2.5 mg/kg) was administered via intraperitoneal injection every 3 days over a period of 7 days before intracerebroventricular injection of 1 μL lipopolysaccharide (10 μg/μL). In addition, astrocyte cultures were treated with 0, 100 or 300 ng/mL lipopolysaccharide, with 200 ng/mL Atsttrin simultaneously. Immunohistochemistry, enzyme-linked immunosorbent assay and real-time reverse transcription-polymerase chain reaction were performed to examine the protein and mRNA levels of inflammatory mediators and to assess activation of the nuclear factor kappa B signaling pathway. Progranulin expression in the brain of wild-type mice and in astrocyte cultures was increased after lipopolysaccharide administration. The protein and mRNA expression levels of tumor necrosis factor-α, interleukin-1β and inducible nitric oxide synthase were increased in the brain of progranulin knockout mice after lipopolysaccharide administration. Atsttrin treatment reduced the lipopolysaccharide-induced increase in the protein and mRNA levels of tumor necrosis factor-α, interleukin-1β, matrix metalloproteinase-3 and inducible nitric oxide synthase in the brain of progranulin knockout mice. Atsttrin also reduced the expression of cyclooxygenase-2, inducible nitric oxide synthase and matrix metalloproteinase 3 mRNA in lipopolysaccharide-treated astrocytes in vitro, and decreased the concentration of tumor necrosis factor a and interleukin-1β in the supernatant. Furthermore, Atsttrin significantly reduced the levels of phospho-nuclear factor kappa B inhibitor a in the brain of lipopolysaccharide-treated progranulin knockout mice and astrocytes, and it decreased the expression of nuclear factor kappa B2 in astrocytes. Collectively, our findings show that the anti-neuroinflammatory effect of Atsttrin involves inhibiton of the nuclear factor kappa B signaling pathway, and they suggest that Atsttrin may have clinical potential in neuroinflammatory therapy. The study was approved by the Animal Ethics Committee of Qilu Hospital of Shandong University, China (approval No. KYLL-2015(KS)-088) on February 10, 2015.