Investigation of the Metabolism of Astragaloside IV in a Puromycin-Damaged Rat Model by UPLC-Q-TOF-MS/MS Analysis

Abstract

Astragaloside IV (AS-IV) has been shown to provide renal protection in various kidney injury models. However, the metabolic profile variation of AS-IV in pathological models in vivo is not well established. This study aims to explore the metabolic pathway of AS-IV in vivo in the classical puromycin aminonucleoside (PAN)-induced kidney injury in a rat model. Twelve Wistar rats were randomly divided into the AS-IV (CA) and the PAN+AS-IV (PA) treatment groups. PAN was injected by a single tail intravenous (i. v.) injection at 5 mg/100 g body weight, and AS-IV was administered intragastrically (i. g.) at 40 mg/kg for 10 days. Fecal samples of these rats were collected, and metabolites of AS-IV were detected by ultra-performance liquid chromatography coupled with quadrupole/time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS) to explore the AS-IV metabolic pathway. The metabolic differences between the AS-IV and PAN+AS-IV groups were compared. A total of 25 metabolites were detected, and deglycosylation, deoxygenation, and methyl oxidation were found to be the main metabolic pathways of AS-IV in vivo. The abundance of most of these metabolites in the PAN+AS-IV group was lower than that in the AS-IV treatment group, and differences for seven of them were statistically significant. Our study indicates that AS-IV metabolism is affected in the PAN-induced kidney injury rat model.

Key words

Astragalus membranaceus - Fabaceae - astragaloside IV - metabolite profiling - UPLC-Q-TOF-MS/MS - puromycin aminonucleoside

Publication History

Received: 26 October 2022

Accepted after revision: 28 September 2023

Article published online:
06 November 2023

• References

• 1 Hill NR, Fatoba ST, Oke JL, Hirst JA, OʼCallaghan CA, Lasserson DS, Hobbs FD. Global prevalence of chronic kidney disease – A systematic review and meta-analysis. PLoS One 2016; 11: e0158765
• 2 Kovesdy CP. Epidemiology of chronic kidney disease: An update 2022. Kidney Int Suppl 2022; 12: 7-11
• 3 Humphreys BD. Mechanisms of renal fibrosis. Annu Rev Physiol 2018; 80: 309-326
• 4 Luyckx VA, Tonelli M, Stanifer JW. The global burden of kidney disease and the sustainable development goals. Bull World Health Organ 2018; 96: 414D-422D
• 5 Pollock C, James G, Garcia Sanchez JJ, Carrero JJ, Arnold M, Lam CSP, Chen HT, Nolan S, Pecoits-Filho R, Wheeler DC. Healthcare resource utilisation and related costs of patients with CKD from the UK: A report from the DISCOVER CKD retrospective cohort. Clin Kidney J 2022; 15: 2124-2134
• 6 Feng H, Zhu X, Tang Y, Fu S, Kong B, Liu X. Astragaloside IV ameliorates diabetic nephropathy in db/db mice by inhibiting NLRP3 inflammasome-mediated inflammation. Int J Mol Med 2021; 48: 164
• 7 Cao Y, Zhang L, Wang Y, Fan Q, Cong Y. Astragaloside IV attenuates renal fibrosis through repressing epithelial-to-mesenchymal transition by inhibiting microRNA-192 expression: In vivo and in vitro studies. Am J Transl Res 2019; 11: 5029-5038
• 8 Xin Y, Li G, Liu H, Ai D. AS-IV protects against kidney IRI through inhibition of NF-κB activity and PUMA upregulation. Int J Clin Exp Med 2015; 8: 18293-18301
• 9 Zhou W, Chen Y, Zhang X. Astragaloside IV alleviates lipopolysaccharide-induced acute kidney injury through down-regulating cytokines, CCR5 and p-ERK, and elevating anti-oxidative ability. Med Sci Monit 2017; 23: 1413-1420
• 10 Lu R, Chen J, Liu B, Lin H, Bai L, Zhang P, Chen D, Li H, Li J, Pang Y, Zhou Y, Zhou J, Wu J. Protective role of astragaloside IV in chronic glomerulonephritis by activating autophagy through PI3K/AKT/AS160 pathway. Phytother Res 2020; 34: 3236-3248
• 11 Zeng Y, Zhang B, Liu X, He L, Wang T, Yu X, Kang Y, Li S. Astragaloside IV alleviates puromycin aminonucleoside-induced podocyte cytoskeleton injury through the Wnt/PCP pathway. Am J Transl Res 2020; 12: 3512-3521
• 12 Shi Z, Jiang J, Zhao D, Xie B, Li Y, Yu C. Effects of astragaloside IV on the pharmacokinetics of metoprolol in rats and its mechanism. Curr Drug Metab 2022; 23: 131-136
• 13 Cheng XD, Wei MG. Profiling the metabolism of astragaloside IV by ultra performance liquid chromatography coupled with quadrupole/time-of-flight mass spectrometry. Molecules 2014; 19: 18881-18896
• 14 Kong S, Ou S, Liu Y, Xie M, Mei T, Zhang Y, Zhang J, Wang Q, Yang B. Surface-enhanced raman spectroscopy analysis of astragalus saponins and identification of metabolites after oral administration in rats by ultrahigh-performance liquid chromatography/quadrupole time-of-flight mass spectrometry analysis. Front Pharmacol 2022; 13: 828449
• 15 Jin Y, Guo X, Yuan B, Yu W, Suo H, Li Z, Xu H. Disposition of astragaloside IV via enterohepatic circulation is affected by the activity of the intestinal microbiome. J Agric Food Chem 2015; 63: 6084-6093
• 16 Zhou RN, Song YL, Ruan JQ, Wang YT, Yan R. Pharmacokinetic evidence on the contribution of intestinal bacterial conversion to beneficial effects of astragaloside IV, a marker compound of astragali radix, in traditional oral use of the herb. Drug Metab Pharmacokinet 2012; 27: 586-597
• 17 Wang YN, Zhang ZH, Liu HJ, Guo ZY, Zou L, Zhang YM, Zhao YY. Integrative phosphatidylcholine metabolism through phospholipase A(2) in rats with chronic kidney disease. Acta Pharmacol Sin 2023; 44: 393-405
• 18 Li H, Zhang H, Yan F, He Y, Ji A, Liu Z, Li M, Ji X, Li C. Kidney and plasma metabolomics provide insights into the molecular mechanisms of urate nephropathy in a mouse model of hyperuricemia. Biochim Biophys Acta Mol Basis Dis 2022; 1868: 166374
• 19 Mo Y, Sun H, Zhang L, Geng W, Wang L, Zou C, Wu Y, Ji C, Liu X, Lu Z. Microbiome-metabolomics analysis reveals the protection mechanism of α-Ketoacid on adenine-induced chronic kidney disease in rats. Front Pharmacol 2021; 12: 657827
• 20 Wang Z, Klipfell E, Bennett BJ, Koeth R, Levison BS, Dugar B, Feldstein AE, Britt EB, Fu X, Chung YM, Wu Y, Schauer P, Smith JD, Allayee H, Tang WH, DiDonato JA, Lusis AJ, Hazen SL. Gut flora metabolism of phosphatidylcholine promotes cardiovascular disease. Nature 2011; 472: 57-63
• 21 Tang WH, Wang Z, Levison BS, Koeth RA, Britt EB, Fu X, Wu Y, Hazen SL. Intestinal microbial metabolism of phosphatidylcholine and cardiovascular risk. N Engl J Med 2013; 368: 1575-1584
• 22 Gupta N, Buffa JA, Roberts AB, Sangwan N, Skye SM, Li L, Ho KJ, Varga J, DiDonato JA, Tang WHW, Hazen SL. Targeted inhibition of gut microbial trimethylamine N-oxide production reduces renal tubulointerstitial fibrosis and functional impairment in a murine model of chronic kidney disease. Arterioscler Thromb Vasc Biol 2020; 40: 1239-1255
• 23 Hu DY, Wu MY, Chen GQ, Deng BQ, Yu HB, Huang J, Luo Y, Li MY, Zhao DK, Liu JY. Metabolomics analysis of human plasma reveals decreased production of trimethylamine N-oxide retards the progression of chronic kidney disease. Br J Pharmacol 2022; 179: 4344-4359
• 24 Janssen EM, Dy SM, Meara AS, Kneuertz PJ, Presley CJ, Bridges JFP. Analysis of patient preferences in lung cancer – estimating acceptable tradeoffs between treatment benefit and side effects. Patient Prefer Adherence 2020; 14: 927-937
• 25 Zhang J, Wu C, Gao L, Du G, Qin X. Astragaloside IV derived from astragalus membranaceus: A research review on the pharmacological effects. Adv Pharmacol 2020; 87: 89-112
• 26 Zhao Y, Li Q, Zhao W, Li J, Sun Y, Liu K, Liu B, Zhang N. Astragaloside IV and cycloastragenol are equally effective in inhibition of endoplasmic reticulum stress-associated TXNIP/NLRP3 inflammasome activation in the endothelium. J Ethnopharmacol 2015; 169: 210-218
• 27 He Q, Han C, Huang L, Yang H, Hu J, Chen H, Dou R, Ren D, Lin H. Astragaloside IV alleviates mouse slow transit constipation by modulating gut microbiota profile and promoting butyric acid generation. J Cell Mol Med 2020; 24: 9349-9361
• 28 Du Y, Zhang Q, Chen GG, Wei P, Tu CY. Pharmacokinetics of astragaloside IV in rats by liquid chromatography coupled with tandem mass spectrometry. Eur J Drug Metab Pharmacokinet 2005; 30: 269-273
• 29 Sun GX, Zhao YY, Miao PP, Yang XY, Miao Q, Li J, Xue BJ, Su J, Zhang YJ. Stability study in biological samples and metabolites analysis of astragaloside IV in rat intestinal bacteria in vitro. Zhongguo Zhong Yao Za Zhi 2014; 39: 4258-4264
• 30 Ma PK, Wei BH, Cao YL, Miao Q, Chen N, Guo CE, Chen HY, Zhang YJ. Pharmacokinetics, metabolism, and excretion of cycloastragenol, a potent telomerase activator in rats. Xenobiotica 2017; 47: 526-537
• 31 Takeuchi DM, Kishino S, Ozeki Y, Fukami H, Ogawa J. Analysis of astragaloside IV metabolism to cycloastragenol in human gut microorganism, bifidobacteria, and lactic acid bacteria. Biosci Biotechnol Biochem 2022; 86: 1467-1475