Wang Daxuan, Yao Xiujuan, Xie Baosong, Chen Yusheng, Lin Changjian
Department of Respiratory and Critical Care Medicine, Fujian Provincial Hospital Fuzhou 350001, Fujian, China.
Provincial School of Clinical Medicine, Fujian Medical University Fuzhou 350001, Fujian, China.
Am J Transl Res. 2021 Nov 15;13(11):12786-12796. eCollection 2021.
As a traditional herbal medicine extracted from the seeds of 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.
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.
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.
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.
鸦胆子油(BJO)是从鸦胆子种子中提取的一种传统草药,已在临床上用于治疗疣、慢性肠胃炎以及包括胃肠道癌和肺癌在内的多种恶性肿瘤。我们最近报道了BJO在治疗肺癌中的抗肿瘤作用及可能的分子机制。然而,BJO是否也具有抗炎作用仍不清楚。
通过实时聚合酶链反应(PCR)和酶联免疫吸附测定(ELISA)检测脂多糖(LPS)处理下巨噬细胞中与肺炎相关的炎症因子。建立LPS诱导的急性肺炎大鼠模型。采用苏木精-伊红(HE)染色检查肺组织的组织病理学变化。实时PCR和ELISA检测也用于检测肺组织中与肺炎相关的炎症因子。
在RAW264.7细胞中,BJO以浓度依赖性方式显著抑制LPS诱导的与肺炎相关的炎症因子(肿瘤坏死因子-α、白细胞介素-1β、白细胞介素-6和白细胞介素-8)的表达和分泌。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核因子-κB(NF-κB)通路并发挥其抗炎功能。蛋白质免疫印迹分析表明,在RAW264.7细胞和肺组织中,LPS刺激后IκB和p65的磷酸化水平显著上调,而IκB的蛋白质水平受到抑制。值得注意的是,在BJO联合处理下,LPS刺激的IκB和p65水平得到有效逆转。p65的表达水平不受LPS和BJO处理的影响。HE染色结果表明,BJO可减少肺组织中炎症细胞的浸润。
BJO可降低肺组织中炎症因子水平,为BJO乳剂作为肺炎辅助治疗提供了理论依据。