Ultra-high-performance liquid chromatography-quadrupole time-of-flight-mass spectrometry-characterized extract of Aerides odorata Lour alleviates paracetamol-induced hepatotoxicity in animal model evidenced by biochemical, molecular, and computational studies

Abstract

Background: Many kinds of orchids have significant health benefits although adequate research on their biological functions is yet to be carried out. This study investigated the paracetamol-induced liver damage–protecting effect of epiphytic Aerides odorata methanol extract (AODE).

Methods: The protective effects of AODE were studied by analyzing its effect on liver function parameters, messenger RNA (mRNA) expression, and tissue histopathological architecture. The results were confirmed by ligand–receptor interaction of molecular docking and multitarget interaction of network pharmacological analyses.

Results: AODE significantly (p < 0.05) minimized the dose-dependent increase in acid phosphatase, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, γ-glutamyl transferase, lactate dehydrogenase, and total bilirubin compared to the reference drug silymarin. Malondialdehyde level decreased, and the antioxidant genes catalase (CAT), superoxide dismutase (SOD), β-actin, paraoxonase-1 (PON1), and phosphofructokinase-1 (PFK-1) were upregulated in AODE-treated paracetamol-intoxicated rats. A total of 376 compounds comprising phenols and flavonoids were identified using ultra-high-performance liquid chromatography-quadrupole time-of-flight-mass spectrometry (UPLC-qTOF-MS). The online toxicity assessment using SwissADME and admetSAR exhibited drug-like, nontoxic, and potential pharmacological properties. Additionally, in silico analysis showed that isoacteoside, one of the identified compounds, exhibited the best docking score (−11.42) with the liver protein human pituitary adenylate cyclase-1 (Protein Data Bank ID: 3N94). Furthermore, network pharmacology analysis identified the top 10 hub genes, namely AKT1 (protein kinase B), CTNNB1 (catenin beta-1), SRC (proto-oncogene c-Src), TNF (tumor necrosis factor), EGFR (epidermal growth factor receptor), HSP90AA1 (heat shock protein 90α), MAPK3 (mitogen-activated protein kinase 3), STAT3 (signal transducer and activator of transcription 3), CASP3 (caspase protein), and ESR1 (estrogen receptor 1), which are responsible for hepatoprotective activity.

Conclusion The findings demonstrate that AODE could be a novel hepatoprotective target in drug-induced liver damage with a further single compound–based animal study.

Keywords

Aerides odorata / animal model / mRNA expression / PCM-induced hepatotoxicity / UPLC-qTOF-MS

References

[1] Khan MZ, Shabbir MI, Saqib Z, et al. Investigation of polyphenol profile, antioxidant activity and hepatoprotective potential of Aconogonon alpinum (All.) Schur roots. Open Chem. 2020;18(1):516-536.

[2] Uddin M, Ali Reza A, Abdullah-Al-Mamun M, et al. Antinociceptive and anxiolytic and sedative effects of methanol extract of Anisomeles indica: an experimental assessment in mice and computer aided models. Front Pharmacol. 2018;9:246.

[3] Soliman MM, Aldhahrani A, Metwally MMJSR. Hepatoprotective effect of Thymus vulgaris extract on sodium nitrite-induced changes in oxidative stress, antioxidant and inflammatory marker expression. Sci Rep. 2021;11(1):1-13.

[4] Moni JNR, Adnan M, Tareq AM, et al. Therapeutic potentials of syzygium fruticosum fruit (seed) reflected into an array of pharmacological assays and prospective receptors-mediated pathways. Life. 2021;11(2):155.

[5] Akhter S, Rahman M, Aklima J, Hasan M, Chowdhury H, Kamirul J. Antioxidative role of Hatikana (Leea macrophylla Roxb.) partially improves the hepatic damage induced by CCl4 in Wistar albino rats. Biomed Res Int. 2015;2015:1-12.

[6] Ayub H, Nadeem M, Mohsin M, et al. A comprehensive review on the availability of bioactive compounds, phytochemicals, and antioxidant potential of plum (Prunus Domestica). Int J Food Prop. 2023;26(1):2388-2406.

[7] Hossen MA, Reza AA, Ahmed AA, et al. Pretreatment of Blumea lacera leaves ameliorate acute ulcer and oxidative stress in ethanol-induced Long-Evan rat: a combined experimental and chemico-biological interaction. Biomed Pharmacother. 2021;135:111211.

[8] Karimi A, Majlesi M, Rafieian-Kopaei M. Herbal versus synthetic drugs; beliefs and facts. J Nephropharmacol. 2015;4(1):27-30.

[9] Hossain MM. Therapeutic orchids: traditional uses and recent advances—an overview. Fitoterapia. 2011;82(2):102-140.

[10] Ahmed AA, Rahman MA, Hossen MA, et al. Epiphytic Acampe ochracea orchid relieves paracetamol-induced hepatotoxicity by inhibiting oxidative stress and upregulating antioxidant genes in in vivo and virtual screening. Biomed Pharmacother. 2021;143:112215.

[11] Chen J-T. Orchid biochemistry. Int J Mol Sci. 2020;21:2338.

[12] Chen J-T. Orchid Biochemistry 2.0. Vol 23. Int J Mol Sci;2022:6823.

[13] Othman MS, Nada A, Zaki HS, Abdel Moneim AE. Effect of Physalis peruviana L. on cadmium-induced testicular toxicity in rats. Biol Trace Elem Res. 2014;159(1):278-287.

[14] Rouf R, Ghosh P, Uzzaman M, et al. Hepatoprotective plants from Bangladesh: a biophytochemical review and future prospect. Evid Based Complement Alternat Med. 2021;2021:1-39.

[15] Gutiérrez P. Orchids: a review of uses in traditional medicine, its phytochemistry and pharmacology. J Med Plant Res. 2010;4:551-558.

[16] Teoh ES. Medicinal Orchids of Asia. Springer;2016.

[17] Sofowora AJI, Nigeria. Medicinal Plants and Traditional Medicine in Africa. Vol 289. Spectrum Books LTD;1993.

[18] Harborne A. Phytochemical Methods a Guide to Modern Techniques of Plant Analysis. Springer Science & Business Media;1998.

[19] Mahmud S, Akhter S, Rahman M, et al. Antithrombotic effects of five organic extracts of Bangladeshi plants in vitro and mechanisms in in silico models. Evid Based Complement Alternat Med. 2015;2015:782742.

[20] Reza AA, Hossain MS, Akhter S, et al. In vitro antioxidant and cholinesterase inhibitory activities of Elatostema papillosum leaves and correlation with their phytochemical profiles: a study relevant to the treatment of Alzheimer’s disease. BMC Complement Altern Med. 2018;18(1):1-8.

[21] Reza AA, Nasrin MS, Alam AK. Phytochemicals, antioxidants, and cholinesterase inhibitory profiles of elatostema papillosum leaves: an alternative approach for management of Alzheimer’s disease. J Neurol Neuromedicine. 2018;3(5):19-22.

[22] Umamaheswari M, Chatterjee TK. In vitro antioxidant activities of the fractions of Coccinia grandis L. leaf extract. Afr J Tradit Complement Altern Med. 2008;5(1):61-73.

[23] Abirami A, Nagarani G, Siddhuraju P. Hepatoprotective effect of leaf extracts from Citrus hystrix and C. maxima against paracetamol induced liver injury in rats. Food Sci Human Wellness. 2015;4:35-41.

[24] Zaoui A, Cherrah Y, Mahassini N, Alaoui K, Amarouch H, Hassar M. Acute and chronic toxicity of nigella sativa fixed oil. Phytomedicine. 2002;9(1):69-74.

[25] Yahya F, Mamat SS, Kamarolzaman MFF, et al. Hepatoprotective activity of methanolic extract of Bauhinia purpurea leaves against paracetamol-induced hepatic damage in rats. Evid Based Complement Alternat Med. 2013;2013:636580.

[26] Högberg J, Larson RE, Kristoferson A, Orrenius S. NADPH-dependent reductase solubilized from microsomes by peroxidation and its activity. Biochem Biophys Res Commun. 1974;56(3):836-842.

[27] Fridovich I. Fundamental aspects of reactive oxygen species, or what’s the matter with oxygen? Ann N Y Acad Sci. 1999;893(1):13-18.

[28] Beers RF, Sizer IW. A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. J Biol Chem. 1952;195(1):133-140.

[29] Moron MS, Depierre JW, Mannervik B. Levels of glutathione, glutathione reductase and glutathione S-transferase activities in rat lung and liver. Biochim Biophys Acta. 1979;582(1):67-78.

[30] Mawa J, Rahman MA, Hashem MA, Juwel HM. Leea macrophylla root extract upregulates the mRNA expression for antioxidative enzymes and repairs the necrosis of pancreatic β-cell and kidney tissues in fructose-fed type 2 diabetic rats. Biomed Pharmacother. 2019;110:74-84.

[31] Barka ZB, Tlili M, Alimi H, et al. Protective effects of edible Rhus tripartita (Ucria) stem extract against ethanol-induced gastric ulcer in rats. J Funct Foods. 2017;30:260-269.

[32] Bulbul MRH, Rahman MA, Rahman MZ, et al. Leea macrophylla (Roxb.) root extract reverses CCl4 induced liver injury through upregulation of antioxidative gene expression: a molecular interaction for therapeutic inception. Adv Tradit Med. 2020;20(1):35-52.

[33] Rose PW, Prlić A, Altunkaya A, et al. The RCSB protein data bank: integrative view of protein, gene and 3D structural information. Nucleic Acids Res. 2017;45(D1): D271-D281.

[34] Kim S, Thiessen PA, Bolton EE, et al. PubChem substance and compound databases. Nucleic Acids Res. 2016;44(D1): D1202-D1213.

[35] Li Z, Wan H, Shi Y, Ouyang P. Personal experience with four kinds of chemical structure drawing software: review on ChemDraw, ChemWindow, ISIS/Draw, and ChemSketch. J Chem Inf Comput Sci. 2004;44(5):1886-1890.

[36] Bell J, Cao Y, Gunn J, et al. PrimeX and the Schrödinger Computational Chemistry Suite of Programs. In International Tables for Crystallography. John Wiley & Sons, Ltd;2012:534-538.

[37] Roos K, Wu C, Damm W, et al. OPLS3e: extending force field coverage for drug-like small molecules. J Chem Theory Comput. 2019;15(3):1863-1874.

[38] Lipinski CA, Lombardo F, Dominy BW, Feeney PJ. Experimental and computational approaches to estimate solubility and permeability in drug discovery and development settings. Adv Drug Deliv Rev. 2001;46(1–3):3-26.

[39] Azzam KA. SwissADME and pkCSM webservers predictors: an integrated online platform for accurate and comprehensive predictions for in silico ADME/T properties of artemisinin and its derivatives. Kompl Ispolz Miner Syra. 2023;325(2):14-21.

[40] Gfeller D, Grosdidier A, Wirth M, Daina A, Michielin O, Zoete V. SwissTargetPrediction: a web server for target prediction of bioactive small molecules. Nucleic Acids Res. 2014;42(W1): W32-W38.

[41] Szklarczyk D, Gable AL, Lyon D, et al. STRING v11: protein–protein association networks with increased coverage, supporting functional discovery in genome-wide experimental datasets. Nucleic Acids Res. 2019;47(D1): D607-D613.

[42] Chin C-H, Chen S-H, Wu H-H, Ho C-W, Ko M-T, Lin C-Y. cytoHubba: identifying hub objects and sub-networks from complex interactome. BMC Syst Biol. 2014;8(4):1-7.

[43] Shannon P, Markiel A, Ozier O, et al. Cytoscape: a software environment for integrated models of biomolecular interaction networks. Genome Res. 2003;13(11):2498-2504.

[44] Huang DW, Sherman BT, Lempicki RA. Bioinformatics enrichment tools: paths toward the comprehensive functional analysis of large gene lists. Nucleic Acids Res. 2009;37(1):1-13.

[45] Van Oss C, Good R, Chaudhury MK. The role of van der Waals forces and hydrogen bonds in “hydrophobic interactions” between biopolymers and low energy surfaces. J Colloid Interface Sci. 1986;111(2):378-390.

[46] Shaik NA, Al-Kreathy HM, Ajabnoor GM, Verma PK, Banaganapalli B. Molecular designing, virtual screening and docking study of novel curcumin analogue as mutation (S769L and K846R) selective inhibitor for EGFR. Saudi J Biol Sci. 2019;26(3):439-448.

[47] Iranshahy M, Iranshahi M, Abtahi SR, Karimi G. The role of nuclear factor erythroid 2-related factor 2 in hepatoprotective activity of natural products: a review. Food Chem Toxicol. 2018;120:261-276.

[48] Xu G-B, Xiao Y-H, Zhang Q-Y, Zhou M, Liao S-G. Hepatoprotective natural triterpenoids. Eur J Med Chem. 2018;145:691-716.

[49] Li S, Tan H-Y, Wang N, et al. The role of oxidative stress and antioxidants in liver diseases. Int J Mol Sci. 2015;16(11):26087-26124.

[50] Rotundo L, Pyrsopoulos N. Liver injury induced by paracetamol and challenges associated with intentional and unintentional use. World J Hepatol. 2020;12(4):125-136.

[51] Chilvery S, Yelne A, Khurana A, et al. Acetaminophen induced hepatotoxicity: an overview of the promising protective effects of natural products and herbal formulations. Phytomed Int J Phytother Phytopharm. 2023;108:154510.

[52] Yoon E, Babar A, Choudhary M, Kutner M, Pyrsopoulos N. Acetaminophen-induced hepatotoxicity: a comprehensive update. J Clin Transl Hepatol. 2016;4(2):131-142.

[53] Zakaria ZA, Kamisan FH, Kek TL, Salleh MZ. Hepatoprotective and antioxidant activities of Dicranopteris linearis leaf extract against paracetamol-induced liver intoxication in rats. Pharm Biol. 2020;58(1):478-489.

[54] El-Sayed YS, Lebda MA, Hassinin M, Neoman SA. Chicory (Cichorium intybus L.) root extract regulates the oxidative status and antioxidant gene transcripts in CCl4-induced hepatotoxicity. PLoS One. 2015;10(3):e0121549.

[55] Sun S, Ge X, Zhu J, Xuan F, Jiang X. Identification and mRNA expression of antioxidant enzyme genes associated with the oxidative stress response in the Wuchang bream (Megalobrama amblycephala Yih) in response to acute nitrite exposure. Comp Biochem Physiol C Toxicol Pharmacol. 2014;159:69-77.

[56] Okokon JE, Simeon JO, Umoh EE. Hepatoprotective activity of the extract of Homalium letestui stem against paracetamol-induced liver injury. Avicenna J Phytomed. 2017;7(1):27-36.

[57] Morales-Ruiz M, Santel A, Ribera J, Jiménez W. The role of Akt in chronic liver disease and liver regeneration. Semin Liver Dis. 2017;37:11-16.

[58] Tornesello ML, Buonaguro L, Tatangelo F, Botti G, Izzo F, Buonaguro FM. Mutations in TP53, CTNNB1 and PIK3CA genes in hepatocellular carcinoma associated with hepatitis B and hepatitis C virus infections. Genomics. 2013;102(2):74-83.

[59] Thapaliya S, Wree A, Povero D, et al. Caspase 3 inactivation protects against hepatic cell death and ameliorates fibrogenesis in a diet-induced NASH model. Dig Dis Sci. 2014;59:1197-1206.

[60] Kurniawan DW, Booijink R, Jajoriya AK, et al. Src kinase as a potential therapeutic target in non-alcoholic and alcoholic steatohepatitis. Clin Transl Discov. 2022;2(1):e18.

[61] Shao F, Deng H, Zhang W, et al. Loss of EGFR contributes to high-fat diet-induced nonalcoholic fatty liver disease. FEBS Lett. 2023;597:1503-1516.

[62] Santen R, Brodie H, Simpson E, Siiteri P, Brodie AJ. History of aromatase: saga of an important biological mediator and therapeutic target. Endocr Rev. 2009;30(4):343-375.

[63] Campbell JS, Argast GM, Yuen SY, Hayes B, Fausto N. Inactivation of p38 MAPK during liver regeneration. Int J Biochem Cell Biol. 2011;43(2):180-188.

[64] Yang Y, Zhang M, Li Z, et al. Identification and cross-validation of autophagy-related genes in cardioembolic stroke. Front Neurol. 2023;14:1097623.

[65] Hao Y-L, Fang H-C, Zhao H-L, et al. The role of microRNA-1 targeting of MAPK3 in myocardial ischemia-reperfusion injury in rats undergoing sevoflurane preconditioning via the PI3K/Akt pathway. Am J Physiol Cell Physiol. 2018;315(3): C380-C388.

[66] Cao HY, Xiao CH, Lu HJ, et al. MiR-129 reduces CDDP resistance in gastric cancer cells by inhibiting MAPK3. Eur Rev Med Pharmacol Sci. 2020;24(22):11468.

[67] Uebi T, Umeda M, Imai T. Estrogen induces estrogen receptor alpha expression and hepatocyte proliferation in the livers of male mice. Genes Cells. 2015;20(3):217-223.

[68] Garcin I, Tordjmann T. Calcium signalling and liver regeneration. Int J Hepatol. 2012;2012:1-6.

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