Saikosaponins and the deglycosylated metabolites exert liver meridian guiding effect through PXR/CYP3A4 inhibition

• Published in: Journal of ethnopharmacology Vol. 279; p. 114344
• Main Authors: Liu, Qiwei, Xue, Yunwen, Liu, Jingjing, Ren, Siqi, Xu, Jie, Yang, Jinni, Xing, Yuanyue, Zhang, Zunjian, Song, Rui
• Format: Journal Article
• Language: English
• Published: Ireland Elsevier B.V 28.10.2021
• Subjects: Bupleurum - chemistry; Carcinoma, Hepatocellular - enzymology; Carcinoma, Hepatocellular - genetics; CYP3A4; Cytochrome P-450 CYP3A - drug effects; Cytochrome P-450 CYP3A - genetics; Cytochrome P-450 CYP3A Inhibitors - administration & dosage; Cytochrome P-450 CYP3A Inhibitors - isolation & purification; Cytochrome P-450 CYP3A Inhibitors - pharmacology; Dose-Response Relationship, Drug; Gene Expression Regulation - drug effects; Hep G2 Cells; Humans; Inhibitory Concentration 50; Liver Neoplasms - enzymology; Liver Neoplasms - genetics; Liver targeting; Meridians; Microsomes, Liver - drug effects; Microsomes, Liver - enzymology; Oleanolic Acid - administration & dosage; Oleanolic Acid - analogs & derivatives; Oleanolic Acid - isolation & purification; Oleanolic Acid - pharmacology; Pregnane X Receptor - drug effects; Pregnane X Receptor - metabolism; PXR; Saikogenin; Saikosaponin; Saponins - administration & dosage; Saponins - isolation & purification; Saponins - pharmacology; Time Factors; Liver targeting; Saikosaponin; PXR; CYP3A4; Saikogenin
• ISSN: 0378-8741; 1872-7573
• DOI: 10.1016/j.jep.2021.114344

Radix Bupleuri (RB), traditionally used to treat inflammatory disorders and infectious diseases, represents one of the most successful and widely used herbal drugs in Asia over the past 2000 years. Being realized the role in regulating metabolism and controlling Yin/Yang, RB is not only chosen specifically for treating liver meridian and the corresponding organs, but also believed to have liver meridian guiding property and help potentiate the therapeutic effects of liver. However, the ingredients in RB with liver meridian guiding property and the underly mechanism have not been comprehensively investigated. Considering the important role of CYP3A4 in first-pass metabolism and the liver exposure of drugs, the present study aimed to determine whether saikosaponins (SSs) and the corresponding saikogenins (SGs) have a role in inhibiting the catalytic activity of CYP3A4 in human liver microsomes and HepG2 hepatoma cells and whether they could suppress CYP3A4 expression by PXR-mediated pathways in HepG2 hepatoma cells. The effect of SSs and SGs on CYP3A4-mediated midazolam1′-hydroxylation activities in pooled human liver microsomes (HLMs) was first studied. Dose-dependent experiments were performed to obtain the half inhibit concentration (IC50) values. HepG2 cells were used to assay catalytic activity of CYP3A4, reporter function, mRNA levels, and protein expression. The inhibitory effects of SSa and SSd on CYP3A4 activity are negligible, while the corresponding SGs (SGF and SGG) have obvious inhibitory effects on CYP3A4 activity, with IC50 values of 0.45 and 1.30 μM. The similar results were obtained from testing CYP3A4 catalytic activity in HepG2 cells, which correlated well with the suppression of the mRNA and protein levels of CYP3A4. Time-dependent testing of CYP3A4 mRNA and protein levels, as well as co-transfection experiments using the CYP3A4 promoter luciferase plasmid, further confirmed that SSs and SGs could inhibit the expression of CYP3A4 at the transcription level. Furthermore, PXR protein expression decreased in a concentration- and time-dependent manner after cells were exposed to SSs and SGs. PXR overexpression and RNA interference experiments further showed that SSs and SGs down-regulate the catalytic activity and expression of CYP3A4 in HepG2 may be mainly through PXR-dependent manner. SSs and SGs inhibit the catalytic activity and expression of CYP3A4 in a PXR-dependent manner, which may be highly related to the liver meridian guiding property of RB. [Display omitted] •Being metabolites, SGs markedly inhibited CYP3A4 enzyme activity rather than their parent compound in HLM and HepG2 cells.•SSs and SGs induced the suppression of CYP3A4 mRNA and protein expression in vitro.•The decrease of CYP3A4 induced by SSs and SGs depends on suppressing PXR expression.