Anti-pulmonary fibrotic activity of salvianolic acid B was screened by a novel method based on the cyto-biophysical properties
Anti-pulmonary fibrotic activity of salvianolic acid B was screened by a novel method based on the cyto-biophysical properties
复制标题
基于细胞生物物理特性的新方法筛选丹酚酸B的抗肺纤维化活性
DOI：
10.1016/j.bbrc.2015.10.127
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发表时间：
2015-12-04
影响因子：
3.1
通讯作者：
Ma, Enlong
中科院分区：
文献类型：
--
作者：
Liu, Miao;Zheng, Mingjing;Ma, Enlong
已经使用各种方法来评价药物的抗纤维化活性。然而，大多数是复杂的，劳动密集型和缺乏效率。本研究旨在建立一种基于生物物理特性的抗肝纤维化药物快速筛选方法。体外A549细胞用转化生长因子-β 1（TGF-β 1）刺激，并通过常规免疫学测定证实纤维形成。同时，利用原子力显微镜（AFM）测量细胞生物物理特性的变化，包括形态，粗糙度和刚度。结果发现，纤维化伴随着细胞生物物理特性的变化。TGF-β 1刺激的A549细胞比对照组明显变长、变粗糙和变硬。然后，分别通过免疫学和生物物理学指标验证了阳性药物N-乙酰-L-半胱氨酸（NAC）对TGF-β 1刺激的A549细胞纤维化的改善作用。主成分分析结果表明，刚度是纤维化发生过程中各项生物物理指标中的主导指标。丹参素B（SalB）是一种天然的抗氧化剂，通过AFM检测到其保护TGF-β 1刺激的A549细胞免于硬化。然后，在博来霉素（BLM）诱导的肺纤维化的大鼠模型上以预防模式提供SalB治疗。结果显示，SalB处理显著改善BLM诱导的肺组织病理学改变，阻断肺组织胶原积聚，降低肺组织中α-SMA表达。这些结果表明SalB具有抗肺纤维化的活性。因此，细胞生物物理特性检测可作为抗肺纤维化药物筛选的一种快速方法。(C)2015 Elsevier Inc. All rights reserved.
Various methods have been used to evaluate anti-fibrotic activity of drugs. However, most of them are complicated, labor-intensive and lack of efficiency. This study was intended to develop a rapid method for anti-fibrotic drugs screening based on biophysical properties. A549 cells in vitro were stimulated with transforming growth factor-beta 1 (TGF-beta 1), and fibrogenesis was confirmed by conventional immunological assays. Meanwhile, the alterations of cyto-biophysical properties including morphology, roughness and stiffness were measured utilizing atomic force microscopy (AFM). It was found that fibrogenesis was accompanied with changes of cellular biophysical properties. TGF-beta 1-stimulated A549 cells became remarkably longer, rougher and stiffer than the control. Then, the effect of N-acetyl-L-cysteine (NAC) as a positive drug on ameliorating fibrogenesis in TGF-beta 1-stimulated A549 cells was verified respectively by immunological and biophysical markers. The result of Principal Component Analysis showed that stiffness was a leading index among all biophysical markers during fibrogenesis. Salvianolic acid B (SalB), a natural anti-oxidant, was detected by AFM to protect TGF-beta 1-stimulated A549 cells against stiffening. Then, SalB treatment was provided in preventive mode on a rat model of bleomycin (BLM) -induced pulmonary fibrosis. The results showed that SalB treatment significantly ameliorated BLM-induced histological alterations, blocked collagen accumulations and reduced alpha-SMA expression in lung tissues. All these results revealed the anti-pulmonary fibrotic activity of SalB. Detection of cyto-biophysical properties were therefore recommended as a rapid method for anti-pulmonary fibrotic drugs screening. (C) 2015 Elsevier Inc. All rights reserved.