Shanxi Provincial Traditional Chinese Medicine Hospital, Taiyuan 030012, China.
Anhui University of Traditional Chinese Medicine, Hefei 230038, China.
J Tradit Chin Med. 2022 Aug;42(4):520-529. doi: 10.19852/j.cnki.jtcm.20220519.001.
To study the effects and mechanism of Shenqihuatan formula (, SQHT) of the transforming growth factor-beta (TGF-β)-stimulated cell processes in airway remodeling.
The current study examined cell viability using a Cell Counting Kit-8 assay. Furthermore, a Transwell assay was conducted to detect the ability of cell migration, and apoptosis was detected via flowcytometry. Western Blot and quantitative real-time polymerase chain reaction (qRT-PCR) were used to determine the expression levels of apoptosis or inflammation-related factors, such as TGF-β, Interleukin-1β (IL-1β), B cell lymphoma 2 (Bcl-2), Bcl-2-Associated X (Bax), Ras homolog gene family, member A (RhoA), recombinant rho associated coiled coil containing protein kinase 1/2 (ROCK1/2), extracellular regulated protein kinases 1/2 (ERK1/2), Snail, and Slug. Finally, the expression levels of matrix metalloproteinase-9 (MMP-9) and Tissue inhibitor of metalloproteinase (TIMP-1) were admeasured by enzyme-linked immuno sorbent assay.
The results demonstrated that SQHT inhibited the viability and migration, as well as the the F-actin formation and cytoskeletal reorganization of airway smooth muscle cells (ASMCs) stimulated by TGF-β. By monitoring the changes of critical regulators in the presence of the formula, it was observed that the expression levels of TGF-β, IL-1β, Bcl-2, RhoA, ROCK1/2, ERK1/2, Snail, and Slug were markedly suppressed, whereas Bax expression exhibited the opposite effect. Compared with a well-characterized RhoA pathway inhibitor, Fasudil, SQHT generated equivalent or even higher inhibitory effects on these processes in ASMCs.
Collectively, these suggested that SQHT can reduce airway inflammation by inhibiting TGF-β-stimulated signaling pathways in ASMCs. These findings may provide a novel remedy for treating ASMC inflammation, which causes thickening and obstruction of the airway in chronic obstructive pulmonary disease.
研究参芪化痰汤（SQHT）对转化生长因子-β（TGF-β）刺激的细胞过程在气道重塑中的作用和机制。
本研究采用细胞计数试剂盒-8 检测细胞活力。此外，通过 Transwell 检测细胞迁移能力，通过流式细胞术检测细胞凋亡。Western Blot 和实时定量聚合酶链反应（qRT-PCR）用于测定凋亡或炎症相关因子的表达水平，如 TGF-β、白细胞介素-1β（IL-1β）、B 细胞淋巴瘤 2（Bcl-2）、Bcl-2 相关 X（Bax）、Ras 同源基因家族成员 A（RhoA）、重组 rho 相关卷曲螺旋蛋白激酶 1/2（ROCK1/2）、细胞外调节蛋白激酶 1/2（ERK1/2）、Snail 和 Slug。最后，通过酶联免疫吸附试验测定基质金属蛋白酶-9（MMP-9）和组织金属蛋白酶抑制剂-1（TIMP-1）的表达水平。
结果表明，SQHT 抑制了 TGF-β刺激的气道平滑肌细胞（ASMCs）的活力和迁移，以及 F-肌动蛋白形成和细胞骨架重排。通过监测公式存在下关键调节剂的变化，观察到 TGF-β、IL-1β、Bcl-2、RhoA、ROCK1/2、ERK1/2、Snail 和 Slug 的表达水平明显受到抑制，而 Bax 的表达则呈现相反的效果。与一种特征明确的 RhoA 通路抑制剂 Fasudil 相比，SQHT 对 ASMCs 中这些过程产生了等效甚至更高的抑制作用。
综上所述，SQHT 可以通过抑制 TGF-β 刺激的 ASMCs 信号通路来减轻气道炎症。这些发现可能为治疗慢性阻塞性肺疾病中引起气道增厚和阻塞的 ASMC 炎症提供一种新的治疗方法。
