Department of Pulmonary Diseases, Wuhan Hospital of Traditional Chinese Medicine, China.
Department of Pharmacy, Wuhan Hospital of Traditional Chinese Medicine, China.
J Toxicol Sci. 2023;48(10):547-556. doi: 10.2131/jts.48.547.
Pulmonary fibrosis is a lethal and progressive pulmonary disorder in human beings. Ephedrine is a compound isolated from Ephedra and plays a regulatory role in inflammatory response. This study focused on the anti-pulmonary fibrosis effect of ephedrine and its potential molecular mechanism. After a mouse model of pulmonary fibrosis was established through bleomycin (BLM) induction, the survival percentage, body weight, and pulmonary index were measured. Hematoxylin-eosin staining and Masson's trichrome staining for lung tissues were performed to observe the pathological alterations. The viability of lung epithelial BEAS-2B cells, intracellular production of reactive oxygen species, and the levels of pro-inflammatory cytokines were examined by cell counting kit-8 assays, 2',7'-dichlorofluorescein diacetate (DCF-DA) staining, and enzyme-linked immunosorbent assay, respectively. Immunofluorescence staining was performed to determine E-cadherin and vimentin expression after BLM or ephedrine treatment. The mRNA and protein levels of cytokeratin-8, E-cadherin, α-SMA, and vimentin were subjected to quantitative polymerase chain reaction and immunoblotting. Experimental results revealed that ephedrine treatment rescued the repressive impact of BLM on BEAS-2B cell viability, and ephedrine inhibited BLM-induced overproduction of reactive oxygen species and inflammatory response in BEAS-2B cells. Additionally, ephedrine suppressed epithelial-mesenchymal transition (EMT) process stimulated by BLM treatment, as demonstrated by the reduced α-SMA and vimentin levels together with the increased cytokeratin-8 and E-cadherin levels in BLM + Ephedrine group. In addition, ephedrine inhibited NF-κB and activated Nrf-2 signaling in BLM-treated BEAS-2B cells. Moreover, ephedrine ameliorated pulmonary fibrosis in BLM-induced mice and improved the survival of model mice. In conclusion, ephedrine attenuates BLM-evoked pulmonary fibrosis by repressing EMT process via blocking NF-κB signaling and activating Nrf-2 signaling, suggesting that ephedrine might become a potential anti-pulmonary fibrosis agent in the future.
肺纤维化是一种致命的、进行性的人类肺部疾病。麻黄碱是从麻黄中分离出来的一种化合物，在炎症反应中发挥调节作用。本研究聚焦于麻黄碱的抗肺纤维化作用及其潜在的分子机制。通过博来霉素（BLM）诱导建立肺纤维化小鼠模型后，测量生存百分比、体重和肺指数。对肺组织进行苏木精-伊红染色和 Masson 三色染色，观察病理改变。通过细胞计数试剂盒-8 测定、2'，7'-二氯荧光素二乙酸酯（DCF-DA）染色和酶联免疫吸附试验分别检测肺上皮 BEAS-2B 细胞活力、细胞内活性氧产生和促炎细胞因子水平。免疫荧光染色用于确定 BLM 或麻黄碱处理后 E-钙粘蛋白和波形蛋白的表达。通过定量聚合酶链反应和免疫印迹检测细胞角蛋白-8、E-钙粘蛋白、α-SMA 和波形蛋白的 mRNA 和蛋白水平。实验结果表明，麻黄碱处理挽救了 BLM 对 BEAS-2B 细胞活力的抑制作用，并且麻黄碱抑制了 BLM 诱导的 BEAS-2B 细胞中活性氧和炎症反应的过度产生。此外，麻黄碱抑制了 BLM 处理刺激的上皮-间充质转化（EMT）过程，BLM+麻黄碱组中α-SMA 和波形蛋白水平降低，细胞角蛋白-8 和 E-钙粘蛋白水平升高。此外，麻黄碱抑制了 BLM 处理的 BEAS-2B 细胞中的 NF-κB 信号并激活了 Nrf-2 信号。此外，麻黄碱改善了 BLM 诱导的小鼠肺纤维化并提高了模型小鼠的存活率。总之，麻黄碱通过阻断 NF-κB 信号并激活 Nrf-2 信号来抑制 EMT 过程，从而减轻 BLM 引起的肺纤维化，这表明麻黄碱在未来可能成为一种潜在的抗肺纤维化药物。
