Screening and identification of potential active components in Ophiopogonis Radix against atherosclerosis by biospecific cell extraction

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2019-12, Vol.1133, p.121817-121817, Article 121817
• Main Authors: Tian, YuShan, Gong, PuYang, Wu, Yan, Chang, Shanquan, Xu, Juntao, Yu, BoYang, Qi, Jin
• Format: Article
• Language: English
• Subjects: Active components; Animals; Antioxidants - analysis; Antioxidants - chemistry; Antioxidants - pharmacology; Atherosclerosis; Biospecific cell extraction; Bone Marrow Cells - cytology; Bone Marrow Cells - metabolism; Cardiovascular Agents - analysis; Cardiovascular Agents - chemistry; Cardiovascular Agents - pharmacology; Cell Survival - drug effects; Cells, Cultured; Chromatography, High Pressure Liquid - methods; Drugs, Chinese Herbal - analysis; Drugs, Chinese Herbal - chemistry; Drugs, Chinese Herbal - pharmacology; Endothelial Cells - cytology; Endothelial Cells - metabolism; Extract of Ophiopogonis Radix; HPLC-Q-TOF-MS/MS; Male; Mice; Ophiopogon; Rats; Rats, Sprague-Dawley; Tandem Mass Spectrometry
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2019.121817

•51 compounds in Ophiopogonis Radix were identified including 22 saponins.•Three primary cells were prepared to approach and simulate the true state.•Biospecific cell extraction performed in pathological state is approach to the fact.•Six multi-effective active constituents were acquired in Ophiopogonis Radix. Atherosclerosis is a chronic disease and an important pathological process associated with cardiovascular disease. Endothelial dysfunction, vascular smooth muscle cells (VSMCs) proliferation and neutrophil activation are involved in the development of atherosclerosis. Ophiopogonis Radix is a common traditional Chinese medicine use to treat cardiovascular diseases, however, its active constituents remain to be elucidated. In this study, primary vascular endothelial cells, primary VSMCs and neutrophils were prepared, and extract of Ophiopogonis Radix (EOR) was investigated to ameliorate H2O2 induced reactive oxygen species (ROS) and nitric oxide (NO) production. The results showed that EOR decreased levels of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1, its protective effects against oxidative damage of endothelia and endothelial dysfunction. Additionally, EOR treatment inhibited oxidized low-density lipoprotein-induced VSMC proliferation, phorbol-12-myristate-13-acetate-mediated ROS production and neutrophil activation, malondialdehyde production, and decreased superoxide dismutase activity and myeloperoxidase release. By HPLC-Q-TOF-MS/MS analysis, 51 compounds in EOR were identified including 22 saponins and 24 homoisoflavonoids. Then biospecific cell extraction and LC-MS technique were employed to screening the antiatherosclerosis active components in Ophiopogonis Radix. After co-cultured with EOR, the multi-effective active constituents including four saponins and two homoisoflavonoids were acquired and subsequently verified to restore properties including endothelial injury, VSMC proliferation and neutrophil activation, indicating that these compounds may be multi-effective active constituents that were responsible for atherosclerosis and the cardiovascular protection of Ophiopogonis Radix.