OBJECTIVE As a traditional herbal medicine extracted from the seeds of Brucea javanica, Brucea javanica oil (BJO) has been clinically used to treat wart, chronic gastroenteritis and a variety of malignant tumors, including gastrointestinal cancer and lung cancer. We have recently reported the anti-tumor role and possible molecular mechanisms of BJO in treatment of lung cancer. However, it remains elusive whether BJO also has an anti-inflammatory effect. METHODS The pneumonia-related inflammatory factors of macrophages under LPS treatment were investigated by real-time PCR and ELISA assays. LPS-induced acute pneumonia rat model was established. Hematoxylin and eosin (HE) examination was performed to detect histopathological changes in the lung tissues. Real-time PCR and ELISA assays were also used to detect the pneumonia-related inflammatory factors in lung tissues. RESULTS LPS-induced expression and secretion of pneumonia-related inflammatory factors (TNF-α, IL-1β, IL-6 and IL-8) were significantly suppressed by BJO in a concentration-dependent manner in RAW264.7 cells. However, BJO did not affect cell proliferation and survival rate. Further mechanistic studies revealed that BJO down-regulated the phosphorylation of IκB and p65, thereby inhibiting NF-κB pathway of macrophages and exerting its anti-inflammatory function. Western blot analysis showed that the phosphorylation levels of IκB and p65 were significantly up-regulated while the protein level of IκB was inhibited upon LPS stimulation in RAW264.7 cells and in lung tissue. Notably, LPS stimulation levels of IκB and p65 were effectively reversed under BJO co-treatment. The expression level of p65 was not influenced by LPS and BJO treatment. HE staining results showed that BJO can reduce the infiltration of inflammatory cells in lung. CONCLUSION BJO can reduce the level of inflammatory factors in lung tissue, which provides a theoretical basis for BJO emulsion as an adjuvant therapy for pneumonia.

中文翻译：

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鸦胆子油通过抑制 NF-κB 活化的抗炎作用。

目的 鸦胆子油（BJO）作为从鸦胆子的种子中提取的传统草药，临床上用于治疗疣、慢性胃肠炎和胃肠道癌、肺癌等多种恶性肿瘤。我们最近报道了 BJO 在肺癌治疗中的抗肿瘤作用和可能的分子机制。然而，BJO 是否也具有抗炎作用仍然难以捉摸。方法采用实时荧光定量PCR和ELISA法研究LPS治疗下巨噬细胞肺炎相关炎症因子。建立LPS诱导的急性肺炎大鼠模型。进行苏木精和伊红（HE）检查以检测肺组织的组织病理学变化。实时荧光定量 PCR 和 ELISA 检测也用于检测肺组织中与肺炎相关的炎症因子。结果 LPS 诱导的肺炎相关炎症因子（TNF-α、IL-1β、IL-6 和 IL-8）的表达和分泌在 RAW264.7 细胞中被 BJO 以浓度依赖性方式显着抑制。然而，BJO 不影响细胞增殖和存活率。进一步的机制研究表明，BJO 下调 IκB 和 p65 的磷酸化，从而抑制巨噬细胞的 NF-κB 通路并发挥其抗炎作用。蛋白质印迹分析表明，在 RAW264.7 细胞和肺组织中，IκB 和 p65 的磷酸化水平显着上调，而 IκB 的蛋白质水平在 LPS 刺激下受到抑制。尤其，IκB 和 p65 的 LPS 刺激水平在 BJO 共同处理下被有效逆转。p65 的表达水平不受 LPS 和 BJO 处理的影响。HE染色结果显示BJO可减少肺内炎症细胞的浸润。结论 BJO可降低肺组织炎症因子水平，为BJO乳剂辅助治疗肺炎提供了理论依据。