Abstract
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.
Highlights
Nonalcoholic fatty liver disease (NAFLD) is one of the most common chronic liver diseases in humans [1]. is disease is characterized by excessive hepatic fat accumulation due to significant ethanol depletion and viral infection [2, 3]
E parameters of the fragmentation energy and impact voltage were optimized to obtain the most abundant relative abundance under the multiple reaction monitoring (MRM) conditions. e results are shown in Figure S1. e MRM chromatograms of the standard solution and sample solutions are shown in Figure 2, which indicates that the
LO2 cells were exposed to FFA to simulate excessive FFA inflow into hepatocytes, resulting in hepatic steatosis. e lipid and TG contents in hepatocytes were considered typical indicators to investigate the lipid regulation effects of different extracts of Hugan Qingzhi formula (HGQZ)
Summary
Nonalcoholic fatty liver disease (NAFLD) is one of the most common chronic liver diseases in humans [1]. is disease is characterized by excessive hepatic fat accumulation due to significant ethanol depletion and viral infection [2, 3]. Nonalcoholic fatty liver disease (NAFLD) is one of the most common chronic liver diseases in humans [1]. In Western countries, the prevalence of NAFLD is between 17% and 33% and significantly increases to 80% in obese individuals [4]. There is no ideal method to control NAFLD. E Hugan Qingzhi formula (HGQZ) is an effective traditional Chinese herbal medicine (CHM) for treating. NAFLD based on the clinical experience of Dr Kuntang Zang, a prestigious traditional Chinese medicine (TCM) doctor at the Southern Medical University of China. Previous studies have shown that HGQZ is effective for treating NAFLD in in vivo and in vitro models [5, 6]. Mechanistic studies have shown that HGQZ acts on adenosine monophosphate-activated protein kinase (AMPK), mitogen-activated protein kinase (MAPK), and peroxisome proliferator-activated receptor (PPAR) signaling pathways
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