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葫芦素 E 通过抑制 NF-κB 核易位在 RAW 264.7 细胞中发挥抗炎作用。

Cucurbitacin E exhibits anti-inflammatory effect in RAW 264.7 cells via suppression of NF-κB nuclear translocation.

机构信息

Department of Immunobiology, Institute of Tissue Transplantation and Immunology, Jinan University, 601 Huangpu Dadao West, Guangzhou 510630, China.

出版信息

Inflamm Res. 2013 May;62(5):461-9. doi: 10.1007/s00011-013-0598-z. Epub 2013 Jan 30.

Abstract

OBJECTIVE

Cucurbitacin E (CuE), a triterpenoid compound isolated from Cucurbitaceae plants, possesses a wide range of biological activities including anti-inflammatory properties. The present study aimed to investigate the anti-inflammatory effect of CuE and the underlying mechanism of action.

METHODS

The anti-inflammatory effect of CuE was evaluated in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Cell proliferation was assessed using a modified MTT assay. Cell cycle distribution was analyzed by propidium iodide staining. The actin cytoskeleton was examined by immunofluorescent staining. The expression of tumor necrosis factor (TNF)-α and interleukin (IL)-1β was determined by intracellular cytokine staining. G-actin level and nuclear factor (NF)-κB nuclear translocation were detected by immunoblotting.

RESULTS

CuE inhibited cell proliferation and induced cell cycle arrest at G2/M phase in RAW 264.7 cells. CuE also suppressed LPS-induced cell spreading and pseudopodia formation. These effects were associated with decreased G-actin level and severe actin aggregation. Moreover, CuE significantly inhibited both TNF-α and IL-1β production in LPS-stimulated RAW 264.7 cells. This was likely mediated by suppressing LPS-induced nuclear translocation of NF-κB, a critical transcription factor responsible for pro-inflammatory cytokine expression.

CONCLUSION

CuE displayed anti-inflammatory effects through suppression of NF-κB nuclear translocation leading to a decreased expression of TNF-α and IL-1β in LPS-stimulated RAW 264.7 cells.

摘要

目的

葫芦素 E(CuE)是从葫芦科植物中分离得到的一种三萜类化合物,具有广泛的生物学活性,包括抗炎特性。本研究旨在探讨 CuE 的抗炎作用及其作用机制。

方法

采用脂多糖(LPS)刺激 RAW 264.7 细胞评估 CuE 的抗炎作用。采用改良 MTT 法评估细胞增殖。碘化丙啶染色分析细胞周期分布。免疫荧光染色观察肌动蛋白细胞骨架。细胞内细胞因子染色检测肿瘤坏死因子(TNF)-α和白细胞介素(IL)-1β的表达。免疫印迹法检测 G 肌动蛋白水平和核因子(NF)-κB 核转位。

结果

CuE 抑制 RAW 264.7 细胞增殖,并诱导细胞周期阻滞于 G2/M 期。CuE 还抑制 LPS 诱导的细胞铺展和伪足形成。这些作用与 G 肌动蛋白水平降低和严重的肌动蛋白聚集有关。此外,CuE 还显著抑制 LPS 刺激的 RAW 264.7 细胞中 TNF-α和 IL-1β的产生。这可能是通过抑制 LPS 诱导的 NF-κB 核转位介导的,NF-κB 是负责促炎细胞因子表达的关键转录因子。

结论

CuE 通过抑制 NF-κB 核转位发挥抗炎作用,从而降低 LPS 刺激的 RAW 264.7 细胞中 TNF-α和 IL-1β的表达。

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