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Phytodolor® (STW 1) 及其成分(杨属植物、白蜡树属植物和泽兰属植物)对人单核细胞/巨噬细胞的抗炎作用。

Anti-inflammatory effects of Phytodolor® (STW 1) and components (poplar, ash and goldenrod) on human monocytes/macrophages.

机构信息

Anatomy und Cell Biology, Department of Medical Cell Biology, Philipps-University of Marburg, Robert-Koch-Str. 8, 35032 Marburg, Germany.

Anatomy und Cell Biology, Department of Medical Cell Biology, Philipps-University of Marburg, Robert-Koch-Str. 8, 35032 Marburg, Germany.

出版信息

Phytomedicine. 2019 May;58:152868. doi: 10.1016/j.phymed.2019.152868. Epub 2019 Feb 18.

Abstract

BACKGROUND

Populus tremula L. (Poplar), Fraxinus excelsior L. (ash) and Solidago virgaurea L. (goldenrod) have been used for medicinal purposes through centuries, to treat pain, fever and inflammation, but their mechanisms of action are still not fully understood. The present study was performed to investigate, whether the herbal medicinal product Phytodolor (STW 1) and its components have anti-inflammatory effects on activated human monocytes and differentiated human macrophages to elucidate their modes of action in comparison with well-known analgesic, non-steroidal anti-inflammatory drug (NSAIDs) as diclofenac.

METHODS

Adherent human monocytes obtained from peripheral blood mononuclear cells (PBMCs) were cultured in serum-free medium and pre-treated with 50-100 µg/ml of diclofenac, STW 1, their components, poplar, ash or goldenrod or its combination (0.05% to 2%). Thereafter, monocytes were activated with 0.1 or 1 µg/ml LPS for 24 h. The intracellular expressions of TNF-α or PTGS2 were determined by cell-based ELISA. Apoptotic cells were identified by YO-PRO-1 staining. Protein or total RNA were isolated to perform SDS-PAGE/Western blot and qRT-PCR analyses. PMA-differentiated human THP-1 macrophages were pre-treated with diclofenac (50 µg/ml) or STW1 (0.1%) and afterwards with LPS (1 µg/ml) and the translocation of the intracellular p62 NF-κB subunit was detected by immunofluorescence.

RESULTS

STW 1 inhibited the intracellular content of TNF-α and PTGS2 protein, as well as of TNF-α and PTGS2 gene expression and induced apoptosis in LPS-activated human monocytes under serum free conditions. Furthermore, STW 1 inhibited the translocation of the p65 subunit of the redox-regulated NF-κB into the nucleus in LPS-activated human macrophages.

CONCLUSION

The present in vitro investigations suggest a significant anti-inflammatory activity of STW 1 and its components by inhibiting pro-inflammatory cytokine as TNF-α and the key enzyme PTGS2 in LPS-activated human monocytes, which is, at least partly mediated through the suppression of NF-κB activation. Our results provide evidence for distinctive anti-inflammatory effects of STW 1 and its components on LPS-activated human monocytes/macrophages and, thus, for the therapeutic use of STW 1 in inflammation and pain related disorders.

摘要

背景

欧洲山杨(Populus tremula L.)、欧洲白蜡(Fraxinus excelsior L.)和一枝黄花(Solidago virgaurea L.)自古以来就被用于治疗疼痛、发热和炎症等医学用途,但它们的作用机制仍不完全清楚。本研究旨在探讨草药药物 Phytodolor(STW 1)及其成分是否对激活的人单核细胞和分化的人巨噬细胞具有抗炎作用,以阐明其作用模式,并与众所周知的镇痛非甾体抗炎药(NSAIDs)如双氯芬酸进行比较。

方法

从外周血单核细胞(PBMCs)中获得贴壁人单核细胞,在无血清培养基中培养,并以 50-100μg/ml 的双氯芬酸、STW 1、其成分、白杨、灰或一枝黄花或其组合(0.05%至 2%)预处理。然后,用 0.1 或 1μg/ml LPS 激活单核细胞 24 小时。通过细胞内 ELISA 测定 TNF-α 或 PTGS2 的细胞内表达。通过 YO-PRO-1 染色鉴定凋亡细胞。分离蛋白质或总 RNA,进行 SDS-PAGE/Western blot 和 qRT-PCR 分析。用 PMA 分化的人 THP-1 巨噬细胞用双氯芬酸(50μg/ml)或 STW1(0.1%)预处理,然后用 LPS(1μg/ml)处理,通过免疫荧光检测细胞内 p62 NF-κB 亚基的易位。

结果

STW 1 在无血清条件下抑制 LPS 激活的人单核细胞中 TNF-α 和 PTGS2 蛋白以及 TNF-α 和 PTGS2 基因表达的细胞内含量,并诱导细胞凋亡。此外,STW 1 抑制 LPS 激活的人巨噬细胞中 p65 亚基向核内易位。

结论

本体外研究表明,STW 1 及其成分通过抑制 LPS 激活的人单核细胞中促炎细胞因子 TNF-α 和关键酶 PTGS2,具有显著的抗炎活性,这至少部分是通过抑制 NF-κB 激活介导的。我们的结果为 STW 1 及其成分对 LPS 激活的人单核细胞/巨噬细胞的独特抗炎作用提供了证据,因此 STW 1 可用于治疗炎症和疼痛相关疾病。

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