Department of Traditional Chinese Medicine, Changhai Hospital, The Second Military Medical University, No.168 Changhai Road, Shanghai, 200433, China.
Seventh People's Hospital of Shanghai University of Traditional Chinese Medicine, No.358 Datong Road, Shanghai, 200137, China.
BMC Complement Med Ther. 2020 Jan 15;20(1):13. doi: 10.1186/s12906-019-2807-y.
Pulmonary fibrosis (PF) is a chronic and progressive interstitial lung disease. Buyang Huanwu Tang (BYHWT), a classical traditional Chinese medicine formula, has been widely utilized for the treatment of PF in China. This present study aimed to explore the mechanism of BYHWT in the treatment of PF in vitro.
TGF-β1 stimulated human alveolar epithelial A549 cells were used as in vitro model for PF. Post the treatment of BYHWT, cell viability was measured by MTT assay, and cell morphology was observed under microscope. The epithelial-to-mesenchymal transition (EMT) markers (E-cadherin, Vimentin) and collagen I (Col I) were detected by western blot, immunofluorescence staining and real-time quantitative polymerase chain reaction. With the co-administration of activators (IGF-1, SC79) and inhibitors (LY294002, MK2206), the effect of BYHWT on PI3K/Akt pathway was analyzed by western blot.
BYHWT inhibited cell growth, and prevented cell morphology changed from epithelial to fibroblasts in TGF-β1 induced A549 cells. BYHWT decreased Vimentin and Col I, while increased E-cadherin at both protein and mRNA levels. Moreover, phosphorylation of PI3K (p-PI3K) and phosphorylation of Akt (p-Akt) were significantly down-regulated by BYHWT in TGF-β1 stimulated A549 cells.
These results indicate that BYHWT suppressed TGF-β1-induced collagen accumulation and EMT of A549 cells by inhibiting the PI3K/Akt signaling pathway. These findings suggest that BYHWT may have potential for the treatment of PF.
肺纤维化（PF）是一种慢性进行性间质性肺疾病。补阳还五汤（BYHWT）是一种经典的中药方剂，在中国被广泛用于治疗 PF。本研究旨在探讨 BYHWT 治疗 PF 的体外作用机制。
采用 TGF-β1 刺激人肺泡上皮 A549 细胞作为 PF 的体外模型。用 BYHWT 处理后，通过 MTT 法测定细胞活力，显微镜下观察细胞形态。通过 Western blot、免疫荧光染色和实时定量聚合酶链反应检测上皮-间充质转化（EMT）标志物（E-钙粘蛋白、波形蛋白）和胶原 I（Col I）。通过共给药激活剂（IGF-1、SC79）和抑制剂（LY294002、MK2206），通过 Western blot 分析 BYHWT 对 PI3K/Akt 通路的影响。
BYHWT 抑制细胞生长，并防止 TGF-β1 诱导的 A549 细胞从上皮细胞向成纤维细胞形态转变。BYHWT 降低了 Vimentin 和 Col I，同时增加了 E-钙粘蛋白的蛋白和 mRNA 水平。此外，BYHWT 显著下调了 TGF-β1 刺激的 A549 细胞中 PI3K 的磷酸化（p-PI3K）和 Akt 的磷酸化（p-Akt）。
这些结果表明，BYHWT 通过抑制 PI3K/Akt 信号通路抑制 TGF-β1 诱导的 A549 细胞胶原积累和 EMT。这些发现表明，BYHWT 可能具有治疗 PF 的潜力。
