胡杨（Tamarix hohenackeri Bunge）通过体外实验对脂多糖激活的小胶质细胞发挥抗炎作用。
Tamarix hohenackeri Bunge exerts anti-inflammatory effects on lipopolysaccharide-activated microglia in vitro.
机构信息
College of Life and Health Sciences, Northeastern University, Shenyang, China.
School of Traditional Chinese Materia Medica, Key Laboratory of Structure-Based Drug Design and Discovery, Shenyang Pharmaceutical University, Ministry of Education, Shenyang, China.
出版信息
Phytomedicine. 2018 Feb 1;40:10-19. doi: 10.1016/j.phymed.2017.12.035. Epub 2017 Dec 28.
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 α (TNF-α), interleukin (IL)-1β and IL-6 were investigated by quantitative real-time PCR (qRT-PCR). The protein levels of phosphorylated of IκBα, ERK and MEK, as well as the cytoplasmic and nuclear NF-κB p65 were tested by Western blot analysis. The translocation of the NF-κB 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-α, IL-1β and IL-6. Western blot analysis showed that compound 7 inhibited the LPS-induced phosphorylation of IκBα and antagonized the LPS-induced reduction of cytoplasmic NF-κB p65 and the increase of nuclear NF-κB p65. Immunofluorescence staining showed that nuclear translocation of NF-κB 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-κB signaling pathway and the MEK/ERK activation in activated microglia. The compound may be potential candidate therapeutic agent for neurodegenerative diseases.
背景
柽柳属植物是肉苁蓉的主要宿主植物，肉苁蓉是一种有价值的中药。它们本身也是传统的药用植物，用于治疗脾病、白癜风和眼部疾病。
目的
本研究旨在研究柽柳 Hohenackeri Bunge 的抗炎作用。
方法
本研究采用脂多糖（LPS）刺激 BV-2 小胶质细胞。用 MTT 法检测细胞活力。用 Griess 法测定一氧化氮（NO）的释放量。用定量实时 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）-1β和 IL-6 的 mRNA 水平。用 Western blot 分析检测磷酸化 IκBα、ERK 和 MEK 的蛋白水平，以及细胞质和核内 NF-κB p65 的蛋白水平。用免疫荧光染色法检测 NF-κB p65 亚基从细胞质向核内的转位。
结果
柽柳 Hohenackeri Bunge 的乙酸乙酯（EtOAc）提取物显著抑制了 NO 的释放。进行了植物化学研究以产生 13 种主要成分。其中，化合物 6、7、10 和 13 被鉴定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 小胶质细胞中 NO 的产生。qRT-PCR 显示，化合物 6 和 7 显著抑制 LPS 诱导的促炎介质基因的转录，包括 iNOS、TNF-α、IL-1β 和 IL-6。Western blot 分析显示，化合物 7 抑制 LPS 诱导的 IκBα 磷酸化，并拮抗 LPS 诱导的细胞质 NF-κB p65 减少和核内 NF-κB p65 增加。免疫荧光染色显示 NF-κB p65 的核转位被化合物 7 抑制。Western blot 分析显示，化合物 7 抑制 LPS 诱导的 ERK 和 MEK 磷酸化。
结论
本研究首次揭示了柽柳属植物的有效抗炎剂。化合物 7 具有较强的抗炎作用，其作用机制可能与其抑制 NF-κB 信号通路和激活小胶质细胞中 MEK/ERK 激活的能力有关。该化合物可能是神经退行性疾病的潜在候选治疗药物。
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