Li Ya-Qun, Xiang Mei-Rong, Rong Rong, Hou Lin, Zhang Cheng-Hua, Zhu Qing-Jun
Shandong University of Traditional Chinese Medicine, Ji'nan 250355, China.
Antiviral Synergistic Innovation Center of Traditional Chinese Medicine in Universities of Shandong Province, Ji'nan 250355, China.
Zhongguo Zhong Yao Za Zhi. 2017 May;42(9):1736-1741. doi: 10.19540/j.cnki.cjcmm.20170222.008.
To study the inhibitory effect of Glehniae Radix petroleum ether part on TGF-β1-induced epithelial mesenchymal transition in non-small cell lung cancer A549 and its possible mechanism. With type Ⅱ epithelial cells of lung cancer A549 as the research object, the experiment was performed in 5 μg•L⁻¹ TGF-β1-induced epithelial mesenchymal transition model,and blank control group, model group and Glehniae Radix petroleum ether group were set up. MTT assay was carried out to detect the effect of petroleum ether extract of Glehniae Radix on the survival of A549 cells. A549 cells induced by TGF-β1(5 μg•L⁻¹) was intervened by different polar parts of Glehniae Radix, Real-time quantitative polymerase chain reaction(RT-qPCR) was used to analyze mRNA expressions of the epithelial mesenchymal transition markers, such as ColⅠ,E-cadherin,Vimentin and α-SMA. Enzyme linked immunosorbent assay(ELISA) was used to detect hydroxyproline(HYP) level. The migration and invasion abilities of cells were detected through wound scratch assay. According to the experimental results, the petroleum ether extract of Glehniae Radix could inhibit the growth of A549 cells in a concentration-dependent manner. Compared with model group, Glehniae Radix petroleum ether part group could effectively inhibit mRNA expressions of ColⅠ,Vimentin and α-SMA, but improve expression of E-cadherin.Glehniae Radix petroleum ether part could reduce the content of hydroxyproline in cells and inhibit the migration of A549 cells.Therefore, the petroleum ether extract of Glehniae Radix can effectively inhibit the occurrence of epithelial mesenchymal transition induced by TGF-β1 induced alveolar epithelial cells, and Glehniae Radix petroleum ether part may be a potential drug for idiopathic pulmonary fibrosis. The mechanism may be achieved through the regulation of ColⅠ, Vimentin, α-SMA and E-cadherin.
研究北沙参石油醚部位对转化生长因子-β1（TGF-β1）诱导的非小细胞肺癌A549上皮-间质转化的抑制作用及其可能机制。以肺癌A549Ⅱ型上皮细胞为研究对象，在5μg•L⁻¹ TGF-β1诱导的上皮-间质转化模型中进行实验，设空白对照组、模型组和北沙参石油醚组。采用MTT法检测北沙参石油醚提取物对A549细胞存活的影响。用北沙参不同极性部位干预TGF-β1（5μg•L⁻¹）诱导的A549细胞，采用实时定量聚合酶链反应（RT-qPCR）分析上皮-间质转化标志物Ⅰ型胶原（ColⅠ）、E-钙黏蛋白（E-cadherin）、波形蛋白（Vimentin）和α-平滑肌肌动蛋白（α-SMA）的mRNA表达。采用酶联免疫吸附测定（ELISA）法检测羟脯氨酸（HYP）水平。通过划痕实验检测细胞的迁移和侵袭能力。实验结果显示，北沙参石油醚提取物能以浓度依赖性方式抑制A549细胞生长。与模型组相比，北沙参石油醚部位组能有效抑制ColⅠ、Vimentin和α-SMA mRNA表达，但提高E-cadherin表达。北沙参石油醚部位可降低细胞内羟脯氨酸含量，抑制A549细胞迁移。因此，北沙参石油醚提取物可有效抑制TGF-β1诱导的肺泡上皮细胞上皮-间质转化的发生，北沙参石油醚部位可能是特发性肺纤维化的潜在药物。其机制可能是通过调控ColⅠ、Vimentin、α-SMA和E-cadherin实现的。
