Key Laboratory of Xin'an Medicine, Ministry of Education, Anhui Province Key Laboratory of R&D of Chinese Medicine, Anhui University of Chinese Medicine, Hefei 230038, China; Institute of Pharmaceutics, Anhui Academy of Chinese Medicine, Hefei 230012, China.
Key Laboratory of Xin'an Medicine, Ministry of Education, Anhui Province Key Laboratory of R&D of Chinese Medicine, Anhui University of Chinese Medicine, Hefei 230038, China; Department of Chinese Medicine, The First Affiliated Hospital of Anhui Medical University, Hefei 230022, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2018 Dec 1;1102-1103:23-33. doi: 10.1016/j.jchromb.2018.10.013. Epub 2018 Oct 17.
Qi-Yu-San-Long Decoction (QYSLD) has been used to treat lung carcinoma for over twenty years in clinical practices, and its curative effect is considered credible. However, the therapeutic mechanism of this effect has not been thoroughly elucidated to date. In this study, a MTT dye reduction assay and DAPI staining were first used to evaluate the cell viability and apoptosis of A549 cells with and without QYSLD-treatment, respectively. The weight/volume of Lewis lung carcinoma (LLC) sarcoma was used to assess the therapeutic effect of QYSLD on LLC mice. Second, an UPLC-QTOF/MS-based untargeted metabolomics method was employed to identify and relatively quantify functional metabolites that were responsible for the intervention effect of QYSLD on LLC. As a result, the MTT dye reduction assay and DAPI staining demonstrated that QYSLD could inhibit the proliferation and induce the apoptosis of A549 cells. The weight/volume test of LLC sarcoma showed that QYSLD could restrain the development of LLC. Next, 21 potential biomarkers that could contribute to the curative mechanism of QYSLD on LLC were screened by the untargeted metabolomics method. The down-regulated metabolites induced by QYSLD included PC(16:0/22:6(4Z,7Z,10Z,13Z,16Z,19Z)), PC(20:2(11Z,14Z)/16:0), PC(22:4(7Z,10Z,13Z,16Z)/14:0), PC(22:5(7Z,10Z,13Z,16Z,19Z)/14:0), arachidonic acid, gamma-glutamylisoleucine, cholesterol sulfate, CL (8:0/10:0/11:0/a-13:0) and CDP-DG (16:0/18:1(11Z)). The up-regulated metabolites were LysoPC(16:0), LysoPC(18:0), LysoPE(18:2(9Z,12Z)/0:0), LysoPE(22:0/0:0), LysoPE(22:1(13Z)/0:0), LysoPE(22:2(13Z,16Z)/0:0), triglylcarnitine, 1‑arachidonoylglycerophosphoinositol, 1‑palmitoylglycerophosphoinositol, 2‑stearoylglycerophosphoinositol, sphingosine 1‑phosphate(d19:1-P) and SM(d18:0/16:1(9Z)). The metabolic pathway analysis revealed that the potential biomarkers were primarily involved in glycerophospholipid metabolism, sphingolipid metabolism, steroid hormone biosynthesis, fatty acid degradation and arachidonic acid metabolism. This study demonstrated that QYSLD has a good antitumor effect and that a UPLC-QTOF/MS-based untargeted metabolomics method is a promising means of elucidating the intervention mechanism of traditional Chinese medicine formulas.
去瘀散结汤（QYSLD）在临床上已用于治疗肺癌超过二十年，其疗效被认为是可信的。然而，其治疗作用的机制至今尚未得到彻底阐明。在这项研究中，首先使用 MTT 染料还原法和 DAPI 染色分别评估了 A549 细胞有无 QYSLD 处理时的细胞活力和细胞凋亡。采用 Lewis 肺癌（LLC）肉瘤的重量/体积来评估 QYSLD 对 LLC 小鼠的治疗效果。其次，采用 UPLC-QTOF/MS 非靶向代谢组学方法鉴定并相对定量与 QYSLD 干预作用相关的功能代谢物。结果表明，MTT 染料还原法和 DAPI 染色表明 QYSLD 可以抑制 A549 细胞的增殖并诱导其凋亡。LLC 肉瘤的重量/体积试验表明 QYSLD 可以抑制 LLC 的发展。接下来，通过非靶向代谢组学方法筛选出 21 个可能与 QYSLD 对 LLC 的治疗机制有关的潜在生物标志物。由 QYSLD 诱导下调的代谢物包括 PC（16:0/22:6（4Z，7Z，10Z，13Z，16Z，19Z））、PC（20:2（11Z，14Z）/16:0）、PC（22:4（7Z，10Z，13Z，16Z）/14:0）、PC（22:5（7Z，10Z，13Z，16Z，19Z）/14:0）、花生四烯酸、γ-谷氨酰异亮氨酸、胆固醇硫酸酯、CL（8:0/10:0/11:0/a-13:0）和 CDP-DG（16:0/18:1（11Z））。上调的代谢物包括溶血磷脂酰胆碱（16:0）、溶血磷脂酰胆碱（18:0）、溶血磷脂酰乙醇胺（18:2（9Z，12Z）/0:0）、溶血磷脂酰乙醇胺（22:0/0:0）、溶血磷脂酰乙醇胺（22:1（13Z）/0:0）、溶血磷脂酰乙醇胺（22:2（13Z，16Z）/0:0）、三肉碱、1-花生四烯酰甘油磷酸肌醇、1-棕榈酰甘油磷酸肌醇、2-硬脂酰甘油磷酸肌醇、鞘氨醇 1-磷酸（d19:1-P）和鞘磷脂（d18:0/16:1（9Z））。代谢途径分析表明，这些潜在的生物标志物主要参与甘油磷脂代谢、鞘脂代谢、甾体激素生物合成、脂肪酸降解和花生四烯酸代谢。本研究表明 QYSLD 具有良好的抗肿瘤作用，并且基于 UPLC-QTOF/MS 的非靶向代谢组学方法是阐明中药方剂干预机制的一种很有前途的手段。
