Tamarix hohenackeri Bunge exerts anti-inflammatory effects on lipopolysaccharide-activated microglia in vitro
Tamarix hohenackeri Bunge exerts anti-inflammatory effects on lipopolysaccharide-activated microglia in vitro
复制标题
柽柳在体外对脂多糖激活的小胶质细胞发挥抗炎作用
DOI：
10.1016/j.phymed.2017.12.035
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发表时间：
2018-02-01
期刊：
影响因子：
7.9
通讯作者：
Li, Ning
中科院分区：
文献类型：
--
作者：
Chen, Ru;Yang, Yanqiu;Li, Ning
背景：红柳属植物是肉苁蓉的主要寄主植物。他们也是传统的药用植物本身，并用于治疗脾脏问题，leukocyte和ocular conditions.Purpose：本研究的目的是调查红柳的抗炎作用。方法：在本研究中，BV-2小胶质细胞被使用和刺激脂多糖（LPS）。使用MTT测定法测试细胞活力。使用Griess测定法测定一氧化氮（NO）的释放。采用实时荧光定量PCR（qRT-PCR）检测诱导型一氧化氮合酶（iNOS）、肿瘤坏死因子α（TNF-α）、白细胞介素（IL）-1 β和IL-6的mRNA水平。Western blot分析I κ B α、ERK和MEK磷酸化蛋白水平以及细胞质和细胞核NF-κ B p65。结果：红柳乙酸乙酯（EtOAc）提取物对NO的释放有明显的抑制作用，经植物化学研究，分离出13种主要成分，其中以NF-kappa B p65亚基为主。其中化合物6、7、10和13为具有抗炎活性的有效成分。这些化合物显著抑制LPS激活的BV-2小胶质细胞产生NO。qRT-PCR显示化合物6和7显著抑制LPS诱导的编码促炎介质的基因的转录，所述促炎介质包括iNOS、TNF-α、IL-1 β和IL-6。Western印迹分析显示，化合物7抑制LPS诱导的I κ B α的磷酸化，并拮抗LPS诱导的细胞质NF-κ B p65的减少和细胞核NF-κ B p65的增加。免疫荧光染色显示化合物7抑制NF-κ B p65的核转位。Western blot分析表明化合物7可抑制LPS诱导的ERK和MEK磷酸化。Hohenackeri化合物7具有较强的抗炎作用，其机制可能与抑制NF-κ B信号通路和活化的小胶质细胞中MEK/ERK活化有关。该化合物可能是神经退行性疾病潜在的候选治疗剂。
Background: Tamarix species are well known as the main host plants of Herba Cistanches, a valuable Traditional Chinese Medicine. They are also traditional medicinal plants themselves and are used to treat spleen problems, leucoderma and ocular conditions.Purpose: The aim of the present study was to investigate the anti-inflammatory effect of Tamarix hohenackeri Bunge.Methods: In the present study, BV-2 microglial cells were used and stimulated with lipopolysaccharide (LPS). Cell viability was tested using the MTT assay. The release of nitric oxide (NO) was determined using the Griess assay. The mRNA level of inducible nitric oxide synthase (iNOS), tumor necrosis factor alpha (TNF-alpha), interleukin (IL)-1 beta and IL-6 were investigated by quantitative real-time PCR (qRT-PCR). The protein levels of phosphory-lated of I kappa B alpha, ERK and MEK, as well as the cytoplasmic and nuclear NF-kappa B p65 were tested by Western blot analysis. The translocation of the NF-kappa B p65 subunit from the cytosol to the nucleus was investigated by immunofluorescence staining.Results: Ethyl acetate (EtOAc) extract of Tamarix hohenackeri Bunge significantly inhibited the release of NO. Phytochemical research was performed to produce 13 main constituents. Among them, compounds 6, 7, 10 and 13 were identified to be the effective components with anti-inflammatory activity. These compounds significantly inhibited the production of NO by LPS-activated BV-2 microglial cells. qRT-PCR showed that compounds 6 and 7 significantly suppressed the LPS-induced transcription of genes encoding pro-inflammatory mediators, including iNOS, TNF-alpha, IL-1 beta and IL-6. Western blot analysis showed that compound 7 inhibited the LPS-induced phosphorylation of I kappa B alpha and antagonized the LPS-induced reduction of cytoplasmic NF-kappa B p65 and the increase of nuclear NF-kappa B p65. Immunofluorescence staining showed that nuclear translocation of NF-kappa B p65 was suppressed by compound 7. Western blot analysis showed that compound 7 inhibited the LPS-induced phosphorylation of ERK and MEK.Conclusion: The present study revealed, for the first time, the effective anti-inflammatory agents from T. Hohenackeri. Compound 7 exerted potent anti-inflammatory effects and its underlying mechanism may be associated with its capacity to inhibit NF-kappa B signaling pathway and the MEK/ERK activation in activated microglia. The compound may be potential candidate therapeutic agent for neurodegenerative diseases.