Integrated analysis of metabolomic and gut microbiota reveals idiosyncratic drug-induced liver injury resulting from the combined administration of bavachin and icariside II

Abstract

Objective: Xianling gubao (XLGB), a widely used Chinese patent medicine for osteoporosis, has garnered significant attention due to its potential to cause liver injury. The constituents Psoraleae Fructus (PF, the dried ripe seeds of Psoralea corylifolia L.) and Epimedii Folium (EF, the dried leaves of various Epimedium species) present in XLGB have been implicated in causing idiosyncratic drug-induced liver injury (IDILI). However, the specific components and mechanisms underlying liver injury related to these tonics remain elusive. This study aims to establish that the combination of bavachin (the primary active compound in PF, and icariside II, the main active compound in EF) induces IDILI in a tumor necrosis factor-α (TNF-α)-mediated mouse model.
Methods: To assess the impact of bavachin and icariside II on the liver in the presence of TNF-α immune stress, an animal model was developed. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) metabolomics technology was employed to identify biomarkers associated with TNF-α-induced IDILI and the combination of bavachin and icariside II. Additionally, 16S rRNA high-throughput sequencing technology was utilized to explore changes in the species composition and relative abundance of gut microbiota. Spearman correlation analysis was conducted to unveil the relationship between gut microbiota and in vivo metabolites.
Results: The study observed that the combined administration of bavachin and icariside II induced liver injury in the TNF-α mediated susceptibility mouse model of IDILI. Under TNF-α stimulation, there was an elevation in levels in mouse livers following bavachin and icariside II administration, while Gly-Tyr, Leu-Gly, and Trp-Ser levels decreased. These differentially expressed metabolites associated with liver injury were predominantly enriched in metabolic pathways such as sphingolipid metabolism, sphingolipid signaling pathway, and necroptosis. it is noteworthy that the gut of mice with liver injury induced by the bavachin and icariside II combination exhibited a significant increase in Bacteroides and Desulfovibrionaceae abundance. Correlation analysis revealed a positive association between Bacteroidaceae and Desulfovibrionaceae with methylcarbamoyl PAF and methyl Indole-3-acetate, while a negative correlation was observed with Gly-Tyr, Leu-Gly, and Trp-Ser.
Conclusions: These findings demonstrated that the combination of bavachin and icariside II increased the risk of IDILI in vivo, providing a promising scientific basis for understanding the component basis of IDILI resulting from the compatibility of EF and PF.

Keywords

Bavachin / Gut microbiota / Icariside II / IDILI / Metabolomics

References

[[1]] Cheng Y, Liu Y, Wang H, et al. A 26-week repeated dose toxicity study of Xian-ling-gu-bao in Sprague-Dawley rats. J Ethnopharmacol. 2013;145:85-93.

[[2]] Wu H, Zhong Q, Wang J, et al. Beneficial effects and toxicity studies of Xian-ling-gu-bao on bone metabolism in ovariectomized rats. Front Pharmacol. 2017;8:273.

[[3]] Li ZR, Cheng LM, Wang KZ, et al. Herbal Fufang Xian Ling Gu Bao prevents corticosteroid-induced osteonecrosis of the femoral head-a first multicentre, randomised, double-blind, placebo-controlled clinical trial. J Orthop Translat. 2018;12:36-44.

[[4]] Wu W, Wang T, Sun B, et al. Xian-Ling-Gu-Bao induced inflammatory stress rat liver injury: inflammatory and oxidative stress playing important roles. J Ethnopharmacol. 2019;239:111910.

[[5]] Li CY, Niu M, Liu YL, et al. Screening for susceptibility-related factors and biomarkers of Xianling Gubao capsule-induced liver injury. Front Pharmacol. 2020;11:810.

[[6]] Giustarini G, Huppelschoten S, Barra M, et al. The hepatotoxic fluoroquinolone trovafloxacin disturbs TNF- and LPS-induced p65 nuclear translocation in vivo and in vitro. Toxicol Appl Pharmacol. 2020;391:114915.

[[7]] Li YG, Hou J, Li SY, et al. Fructus Psoraleae contains natural compounds with potent inhibitory effects towards human carboxylesterase 2. Fitoterapia. 2015;101:99-106.

[[8]] Ma H, He X, Yang Y, et al. The genus Epimedium: an ethnopharmacological and phytochemical review. J Ethnopharmacol. 2011;134:519-541.

[[9]] Chopra B, Dhingra AK, Dhar KL. Psoralea corylifolia L.(Buguchi) - folklore to modern evidence: review. Fitoterapia. 2013;90:44-56.

[[10]] Chen XJ, Tang ZH, Li XW, et al. Chemical constituents, quality control, and bioactivity of Epimedii Folium (Yinyanghuo). Am J Chin Med. 2015;43:783-834.

[[11]] Gao Y, Wang Z, Tang J, et al. New incompatible pair of TCM: Epimedii Folium combined with Psoraleae Fructus induces idiosyncratic hepatotoxicity under immunological stress conditions. Front Med. 2020;14:68-80.

[[12]] Wang Z, Xu G, Wang H, et al. Icariside II, a main compound in Epimedii Folium, induces idiosyncratic hepatotoxicity by enhancing NLRP3 inflammasome activation. Acta Pharm Sin B. 2020;10:1619-1633.

[[13]] Johnson CH, Ivanisevic J, Siuzdak G. Metabolomics: beyond biomarkers and towards mechanisms. Nat Rev Mol Cell Biol. 2016;17:451-459.

[[14]] Wang M, Chen L, Liu D, et al. Metabolomics highlights pharmacological bioactivity and biochemical mechanism of traditional Chinese medicine. Chem Biol Interact. 2017;273:133-141.

[[15]] Liao Y, Wang C, Gao Z, et al. Anti-obesity mechanism of Ganpu tea revealed by microbiome, metabolome and transcriptome analyses. Food Chem. 2023;412:135048.

[[16]] Albillos A, de Gottardi A, Rescigno M. The gut-liver axis in liver disease: pathophysiological basis for therapy. J Hepatol. 2020;72:558-577.

[[17]] Wang R, Tang R, Li B, et al. Gut microbiome, liver immunology, and liver diseases. Cell Mol Immunol. 2021;18:4-17.

[[18]] Qin N, Xu G, Wang Y, et al. Bavachin enhances NLRP3 inflammasome activation induced by ATP or nigericin and causes idiosyncratic hepatotoxicity. Front Med. 2021;15:594-607.

[[19]] Próchnicki T, Latz E. Inflammasomes on the crossroads of innate immune recognition and metabolic control. Cell Metab. 2017;26:71-93.

[[20]] Quintás G, Martínez-Sena T, Conde I, et al. Metabolomic analysis to discriminate drug-induced liver injury (DILI) phenotypes. Arch Toxicol. 2021;95:3049-3062.

[[21]] Dennis EA, Norris PC. Eicosanoid storm in infection and inflammation. Nat Rev Immunol. 2015;15:511-523.

[[22]] Hollie NI, Cash JG, Matlib MA, et al. Micromolar changes in lysophosphatidylcholine concentration cause minor effects on mitochondrial permeability but major alterations in function. Biochim Biophys Acta. 2014;1841:888-895.

[[23]] Kakisaka K, Cazanave SC, Fingas CD, et al. Mechanisms of lysophosphatidylcholine-induced hepatocyte lipoapoptosis. Am J Physiol Gastrointest Liver Physiol. 2012;302:G77-G84.

[[24]] Glunde K, Jacobs MA, Bhujwalla ZM. Choline metabolism in cancer: implications for diagnosis and therapy. Expert Rev Mol Diagn. 2006;6:821-829.

[[25]] Li L, Wang H, Jones JW. Sphingolipid metabolism as a marker of hepatotoxicity in drug-induced liver injury. Prostaglandins Other Lipid Mediat. 2020;151:106484.

[[26]] Nojima H, Freeman CM, Gulbins E, et al. Sphingolipids in liver injury, repair and regeneration. Biol Chem. 2015;396:633-643.

[[27]] Kwong EK, Li X, Hylemon PB, et al. Sphingosine kinases/sphingosine 1-phosphate signaling in hepatic lipid metabolism. Curr Pharmacol Rep. 2017;3:176-183.

[[28]] Bauer KC, Littlejohn PT, Ayala V, et al. Nonalcoholic fatty liver disease and the gut-liver axis: exploring an undernutrition perspective. Gastroenterology.2022;162:1858-1875.e2.

[[29]] Woodhouse CA, Patel VC, Singanayagam A, et al. Review article: the gut microbiome as a therapeutic target in the pathogenesis and treatment of chronic liver disease. Aliment Pharmacol Ther. 2018;47:192-202.

[[30]] Zhu L, Baker SS, Gill C, et al. Characterization of gut microbiomes in nonalcoholic steatohepatitis (NASH) patients: a connection between endogenous alcohol and NASH. Hepatology. 2013;57:601-609.

[[31]] Shen B, Wang J, Guo Y, et al. Dextran sulfate sodium salt-induced colitis aggravates gut microbiota dysbiosis and liver injury in mice with non-alcoholic steatohepatitis. Front Microbiol. 2021;12:756299.

[[32]] Boursier J, Mueller O, Barret M, et al. The severity of nonalcoholic fatty liver disease is associated with gut dysbiosis and shift in the metabolic function of the gut microbiota. Hepatology. 2016;63:764-775.

[[33]] Panasevich MR, Meers GM, Linden MA, et al. High-fat, high-fructose, high-cholesterol feeding causes severe NASH and cecal microbiota dysbiosis in juvenile Ossabaw swine. Am J Physiol Endocrinol Metab. 2018;314:E78-E92.

[[34]] Zhang X, Coker OO, Chu ES, et al. Dietary cholesterol drives fatty liver-associated liver cancer by modulating gut microbiota and metabolites. Gut. 2021;70:761-774.

[[35]] Wang Q, Wang C, Abdullah, et al. Hydroxytyrosol alleviates dextran sulfate sodium-induced colitis by modulating inflammatory responses, intestinal barrier, and microbiome. J Agric Food Chem. 2022;70:2241-2252.

[[36]] Lordan R, Tsoupras A, Zabetakis I, et al. Forty years since the structural elucidation of platelet-activating factor (PAF): historical, current, and future research perspectives. Molecules. 2019;24:4414.

[[37]] Marrache AM, Gobeil F Jr, Bernier SG, et al. Proinflammatory gene induction by platelet-activating factor mediated via its cognate nuclear receptor. J Immunol. 2002;169:6474-6481.

[[38]] Yin H, Shi A, Wu J. Platelet-activating factor promotes the development of non-alcoholic fatty liver disease. Diabetes Metab Syndr Obes. 2022;15:2003-2030.

[[39]] Pavlova T, Vidova V, Bienertova-Vasku J, et al. Urinary intermediates of tryptophan as indicators of the gut microbial metabolism. Anal Chim Acta. 2017;987:72-80.

[[40]] Paulusma CC, Lamers WH, Broer S, et al. Amino acid metabolism, transport and signalling in the liver revisited. Biochem Pharmacol. 2022;201:115074.

[[41]] Hu C, Li H, Wu L, et al. Metabolic profiling of 19 amino acids in triptolide-induced liver injured rats by gas chromatography-triple quadrupole mass spectrometry. Hum Exp Toxicol. 2021;40:1685-1697.

[[42]] Men L, Wang Z, Gou M, et al. Metabolomics and targeted amino acid analysis reveal the liver protective effect of arginyl-fructosyl-glucose from red ginseng on acute liver injury in mice. J Funct Foods. 2023;103:105473

[[43]] Marsh DC, Vreugdenhil PK, Mack VE, et al. Glycine protects hepatocytes from injury caused by anoxia, cold ischemia and mitochondrial inhibitors, but not injury caused by calcium ionophores or oxidative stress. Hepatology. 1993;17:91-98.

[[44]] Barakat HA, Hamza AH. Glycine alleviates liver injury induced by deficiency in methionine and or choline in rats. Eur Rev Med Pharmacol Sci. 2012;16:728-736.

[[45]] Hua H, Xu X, Tian W, et al. Glycine alleviated diquat-induced hepatic injury via inhibiting ferroptosis in weaned piglets. Anim Biosci. 2022;35:938-947.

RIGHTS & PERMISSIONS

2024 Acupuncture and Herbal Medicine