Department of Respiratory Medicine, The Second Hospital of Shandong University, Jinan, 250033, Shandong, PR China; Department of Respiratory Medicine, Central Hospital of Tai'an of Shandong Province, Tai'an, 271000, Shandong, PR China.
Department of Respiratory Medicine, The Second Hospital of Shandong University, Jinan, 250033, Shandong, PR China.
Biomed Pharmacother. 2019 Nov;119:109387. doi: 10.1016/j.biopha.2019.109387. Epub 2019 Sep 2.
To observe the effect of astragaloside ASV (ASV) on silicosis fibroblasts, and further investigate its regulatory mechanism on TGF-β1/Smad3 signaling pathway.
Silica-induced rats model was established in this study. RT-qPCR was performed to detect α-SMA, Collagen I, Collagen III, Smad2, Smad3 and Smad7 expression. Immunofluorescence was conducted to detect α-SMA, Collagen I, Collagen III and p-Smad3 protein and the nucleoplasmic distribution of p-Smad3.Western-blotting was performed to detect the protein of Smad2, p-Smad2, Smad3, p-Smad3 and Smad7.
20 μg/mL ASV could effectively reduce the expression of α-SMA, Collagen I, Collagen III. TGF-β1 stimulated the proliferation of fibroblasts, promoted phosphorylation of Smad2 and Smad3, and down-regulated Smad7 expression. Among them, continuous phosphorylation of Smad3 is a major factor in causing fibrosis. Besides, ASV can inhibit silica-induced lung fibroblast fibrosis through TGF-β1/Smad3 signaling pathway, thereby inhibiting the formation of silicosis.
ASV could inhibit the expression of collagen in fibroblasts and the transformation to myofibroblasts, and has an anti-silicosis fibrosis effect, which may be related to the continuous phosphorylation of Smad3 in the TGF-β1/Smad signaling pathway.
观察黄芪甲苷（ASV）对矽肺成纤维细胞的作用，进一步探讨其对 TGF-β1/Smad3 信号通路的调控机制。
本研究建立了矽肺大鼠模型。采用 RT-qPCR 检测 α-SMA、Collagen I、Collagen III、Smad2、Smad3 和 Smad7 的表达。免疫荧光法检测 α-SMA、Collagen I、Collagen III 和 p-Smad3 蛋白及 p-Smad3 的核质分布。Western-blotting 检测 Smad2、p-Smad2、Smad3、p-Smad3 和 Smad7 的蛋白表达。
20μg/mL ASV 可有效降低 α-SMA、Collagen I、Collagen III 的表达。TGF-β1 刺激成纤维细胞增殖，促进 Smad2 和 Smad3 磷酸化，下调 Smad7 表达。其中，Smad3 的持续磷酸化是导致纤维化的主要因素。此外，ASV 可通过 TGF-β1/Smad3 信号通路抑制矽肺肺成纤维细胞纤维化，从而抑制矽肺的形成。
ASV 可抑制成纤维细胞中胶原的表达和向肌成纤维细胞的转化，具有抗矽肺纤维化作用，可能与 TGF-β1/Smad 信号通路中 Smad3 的持续磷酸化有关。
