He Fan, Xiao Chun-Xin, Wang Can-Jian, Liang Jie, Cheng Qi-Qing, Zhang Li, Zhou Ben-Jie, Zhou Hua
State Key Laboratory of Quality Research in Chinese Medicine and Faculty of Chinese Medicine, Macau University of Science and Technology, Taipa, Macao, China.
Department of Pharmacy, The Seventh Affiliated Hospital, Sun Yat-sen University, Shenzhen, China.
Evid Based Complement Alternat Med. 2020 Nov 12;2020:1594350. doi: 10.1155/2020/1594350. eCollection 2020.
Nonalcoholic fatty liver disease (NAFLD) is a prevalent chronic liver disease. The Hugan Qingzhi formula (HGQZ) has been proven effective in treating NAFLD through clinical and pharmacological mechanism studies. A screening study of the chemical components was carried out to better control the quality of this formula. Current research has combined biological activity assessment with chemical analysis to screen and identify the bioactive compounds in HGQZ for use as potential quality markers (Q-markers) to control the quality of this herbal product. The HGQZ extracted by three different solvents was evaluated in a free fatty acid-induced hepatic steatosis LO2 cell model. Simultaneously, the twelve major chemical constituents of these extracts were quantitatively measured by ultrahigh-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UHPLC-QQQ-MS/MS). Extraction with 50% ethanol showed the most potent lipid-lowering effect in steatosis LO2 cells and the highest extraction rate of major chemical constituents. Correlation analysis was used to establish the relationship between the biological activities and chemical characteristics of these extracts. The results showed that the contents of typhaneoside, hyperoside, isoquercitrin, isorhamnetin-3-O-neohesperidoside, notoginsenoside R1, and alisol B 23-acetate were positively correlated to the lipid-lowering effect. The subsequent bioassay confirmed that typhaneoside, isoquercitrin, and alisol B 23-acetate played the role of reducing the lipid effect. In conclusion, 50% of ethanol extraction produced the most active extract of HGQZ. Typhaneoside, isoquercitrin, and alisol B 23-acetate could be considered potential Q-markers for the quality control of HGQZ.
非酒精性脂肪性肝病（NAFLD）是一种常见的慢性肝病。护肝清脂方（HGQZ）已通过临床和药理机制研究证明对治疗NAFLD有效。为了更好地控制该方剂的质量，开展了化学成分筛选研究。目前的研究将生物活性评估与化学分析相结合，以筛选和鉴定HGQZ中的生物活性化合物，用作潜在的质量标志物（Q-标志物）来控制该草药产品的质量。在游离脂肪酸诱导的肝脂肪变性LO2细胞模型中评估了用三种不同溶剂提取的HGQZ。同时，采用超高效液相色谱-三重四极杆质谱联用（UHPLC-QQQ-MS/MS）对这些提取物的12种主要化学成分进行了定量测定。50%乙醇提取在脂肪变性LO2细胞中显示出最有效的降脂作用，且主要化学成分的提取率最高。采用相关性分析建立这些提取物的生物活性与化学特征之间的关系。结果表明，巴戟天苷、金丝桃苷、异槲皮苷、异鼠李素-3-O-新橙皮苷、三七皂苷R1和泽泻醇B 23-乙酸酯的含量与降脂作用呈正相关。随后的生物测定证实，巴戟天苷、异槲皮苷和泽泻醇B 23-乙酸酯具有降低脂质的作用。综上所述，50%乙醇提取得到的HGQZ提取物活性最强。巴戟天苷、异槲皮苷和泽泻醇B 23-乙酸酯可被视为HGQZ质量控制的潜在Q-标志物。
