Protective effects of turmeric extract, curcumin, demethoxycurcumin, bis-demethoxycurcumin, and ar-turmerone from Curcuma longa L. rhizomes on acetaminophen-induced hepatotoxicity in HepG2 cells

Abstract

Objective: To assess the effects of turmeric extract and its compounds on oxidative stress, inflammation, and apoptosis in acetaminophen-induced liver injury.

Methods: HepG2 cells were administered with acetaminophen (40 mM) to induce hepatotoxicity, followed by treatment with turmeric extract and its isolated compounds including curcumin, demethoxycurcumin, bis-demethoxycurcumin and ar-turmerone at 5, 25, and 125 μg/mL. IL-1β, IL-6, and IL-10 levels were quantified with ELISA kits. Further, qRT-PCR was used to analyze the mRNA expression of JNK, Casp-9, and Casp-3. Meanwhile, the levels of nitric oxide and lactate dehydrogenase were analyzed using colorimetric assay.

Results: Acetaminophen administration caused an increase in the levels of lactate dehydrogenase, nitric oxide, IL-1β, IL-6, and the mRNA expression of JNK, Casp-9, and Casp-3 in HepG2 cells while reducing IL-10 levels. Treatment with turmeric extract, curcumin, demethoxycurcumin, bis-demethoxycurcumin, and ar-turmerone lowered IL-1β, IL-6, nitric oxide, and lactate dehydrogenase levels, downregulated the mRNA expression of JNK, Casp-9, and Casp-3, and increased IL-10 levels.

Conclusions: Turmeric extract and its compounds have significant hepatoprotective activity and could be further explored for the treatment of liver damage.

Keywords

Acetaminophen / Apoptosis / Curcuma longa / Curcumin / Hepatotoxicity / Inflammation / Oxidative stress

Conflict of interest statement

The authors declare that no competing interests exist.

Acknowledgments

The authors express their gratitude to Maranatha Christian University, Bandung, Indonesia for grant of Productive Lecturer Research (No: 011/SK/ADD/UKM/IV/2024) and R.B.V.R.R. Women’s College, Narayanaguda, Hyderabad, Telangana, India for financial support. Appreciation is also extended to PT. Fathonah Amanah Shidiq Tabligh, Depok, West Java, Indonesia, for helping the production of TE, and Aretha Medika Utama, Bandung, Indonesia, for their contribution to the research methodology, facilities, and invaluable assistance.

Funding

This research was funded by Maranatha Christian University, Bandung, Indonesia for Productive Lecturer Research under grant number: 011/SK/ADD/UKM/IV/2024.

Data availability statement

The data supporting the findings of this study are available from the corresponding author upon request.

Authors’ contributions

WW conceptualized, supervised the research project, contributed to manuscript drafting and revision, and secured funding for the project. DRL, DKP, and DR were responsible for the design and execution of the cell-based experiments and data analysis. SRK and JAD performed compound isolation and characterization. DP contributed to the methodology and interpretation of the gene expression data. NSMD, AFS, and RA conducted ELISA and colorimetric assays and performed statistical analyses. DSH was involved in data curation, graphical illustrations, and manuscript editing. ME provided critical revisions and ensured the scientific integrity of the manuscript. All authors have read and approved the final version of the manuscript.

Publisher’s note

The Publisher of the Journal remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

References

[1] Hosack T, Damry D, Biswas S. Drug-induced liver injury: A comprehensive review. Therap Adv Gastroenterol 2023;16. doi: 10.1177/17562848231163410.

[2] Teschke R, Danan G. Idiosyncratic DILI and RUCAM under one hat: The global view. Livers 2023; 3(3):397-433. doi: 10.3390/livers3030030.

[3] Björnsson ES. Epidemiology, predisposing factors, and outcomes of drug-induced liver injury. Clin Liver Dis 2019; 24(1):1-10. doi: 10.1016/j.cld.2019.08.002.

[4] Shen T, Liu Y, Shang J, Xie Q, Li J, Yan M, et al. Incidence and etiology of drug-induced liver injury in Mainland China. Gastroenterology 2019; 156(8):2230-41.e11. doi: 10.1053/j.gastro.2019.02.002.

[5] Li X, Lao R, Lei J, Chen Y, Zhou Q, Wang T, et al. Natural products for acetaminophen-induced acute liver injury: A review. Molecules 2023; 28(23). doi: 10.3390/molecules28237901.

[6] Ramachandran A, Jaeschke H. Oxidant stress and acetaminophen hepatotoxicity: Mechanism-based drug development. Antioxid Redox Signal 2021; 35(9):718-733. doi: 10.1089/ars.2021.0102.

[7] Kurian R, Steen LT, Wang H. Oral bioavailability and drug delivery: From basics to advanced concepts and applications. United State: John Wiley & Sons; 2023, p. 189-212.

[8] Jollow D. Acetaminophen-induced hepatic necrosis: A reminiscence. Drug Metab Dispos 2024; 52(8):707-711. doi: 10.1124/dmd.123.001278.

[9] Mohammed IJ, Sarhat ER, Hamied MAS, Sarhat TR. Assessment of salivary interleukin (IL)-6, IL-10, oxidative stress, antioxidant status, pH, and flow rate in dental caries experience patients in Tikrit Province. Sys Rev Pharm 2021; 12(1):55-59. doi: 10.31838/srp.2021.1.10.

[10] Widowati W, Prahastuti S, Hidayat M, Hasianna ST, Wahyudianingsih R, Eltania TF, et al. Detam 1 black soybean against cisplatin-induced acute renal failure on rat model via antioxidant, anti-inflammatory and anti-apoptosis potential. J Tradit Complement Med 2022;12:426-435. doi: 10.1016/j.jtcme.2022.01.004.

[11] Kumar M, Kaur S, Kaur S. c-Jun N-terminal Kinase (JNK), p38, and caspases: Promising therapeutic targets for the regulation of apoptosis in cancer cells by phytochemicals. Curr Cancer Ther Rev 2024; 20(2):200-211. doi: 10.2174/1573394719666230817094831.

[12] Chang L, Xu D, Zhu J, Ge G, Kong X, Zhou Y. Herbal therapy for the treatment of acetaminophen-associated liver injury: Recent advances and future perspectives. Front Pharmacol 2020;11:313. doi: 10.3389/fphar.2020.00313.

[13] Ballester P, Cerda B, Arcusa R, García-Muñoz AM, Marhuenda J, Zafrilla P. Antioxidant activity in extracts from Zingiberaceae family: Cardamom, turmeric, and ginger. Molecules 2023; 28(10):4024-4044. doi: 10.3390/molecules28104024.

[14] Mirihagalla MKPN, Fernando KMC. Medicinal plants use for home remedies in Sri Lanka: A review. Int J Minor Fruits Med Arom Plants 2021; 7(1):29-39. doi: 10.53552/ijmfmap.2021.v07ii02.002.

[15] Ranjbar R, Bagheri H, Ghasemi F, Guest PC, Sahebkar A. Effects of curcumin and its analogues on infectious diseases. Adv Exp Med Biol 2021;1291:75-101. doi: 10.1007/978-3-030-56153-6_5.

[16] Ibrahim J, Kabiru AY, Abdulrasheed-Adeleke T, Lawal B, Adewuyi AH. Antioxidant and hepatoprotective potentials of curcuminoid isolates from turmeric (Curcuma longa) rhizome on CCl₄-induced hepatic damage in Wistar rats. J Taibah Univ Sci 2020; 14(1):908-915. doi: 10.1080/16583655.2020.1790928.

[17] Choi Y, Ban I, Lee H, Baik MY, Kim W. Puffing as a novel process to enhance the antioxidant and anti-inflammatory properties of Curcuma longa L. (turmeric). Antioxidants 2019; 8(11):506-517. doi: 10.3390/antiox8110506.

[18] Widowati W, Darsono L, Lucianus J, Setiabudi E, Obeng SS, Stefani S, et al. Butterfly pea flower (Clitoria ternata L.) extract displayed antidiabetic effect through antioxidant, anti-inflammatory, lower hepatic GSK-3β and pancreatic glycogen on diabetes mellitus and dyslipidemia rat. J King Saud Univ Sci 2023; 35. doi: 10.1016/j.jksus.2023.102579.

[19] Widowati W, Jasaputra K, Gunawan KY, Kusuma HSW, Arumwardana S, Wahyuni CD, et al. Turmeric extract potential inhibit inflammatory marker in LPS-stimulated macrophage cells. Int J Appl Pharm 2021;13:7-11. doi: 10.22159/IJAP.2021.V13S3.01.

[20] Ginting CN, Lister INE, Girsang E, Mutia MS, Lubis YEP, Amalia A, et al. Ficus elastica leaf extract effect toward IL-10 and TNF-α as a preeclampsia model on hypoxia-induced EA.hy926. J Math Fund Sci 2021; 53(3):487-496. doi: 10.5614/j.math.fund.sci.2021.53.3.10.

[21] Widowati W, Kusuma HSW, Arumwardana S, Afifah E, Wahyuni CD, Wijayanti CR, et al. Corilagin potential in inhibiting oxidative and inflammatory stress in LPS-induced murine macrophage cell lines (RAW 264.7). Iran J Basic Med Sci 2021;24:1656-1665. doi: 10.22038/IJBMS.2021.59348.13174.

[22] Lister INE, Ginting CN, Girsang E, Amansyah A, Chiuman L, Yanti NLWE, et al. Hepatoprotective effect of eugenol on acetaminophen-induced hepatotoxicity in HepG 2 cells. J Phys Conf Ser 2019; 1374(1). doi: 10.1088/1742-6596/1374/1/012009.

[23] Girsang E, Ginting CN, Lister INE, Gunawan KY, Widowati W. Anti-inflammatory and antiaging properties of chlorogenic acid on UV-induced fibroblast cell. Peer J 2021; 9. doi: 10.7717/peerj.11419.

[24] Widowati W, Wargasetia TL, Rahardja F, Gunanegara RF, Priyandoko D, Gondokesumo ME, et al. hWJMSCs inhibit inflammation and apoptosis in an ARDS cell model. J Taibah Univ Med Sci 2023;18:1519-1526. doi: 10.1016/j.jtumed.2023.06.007.

[25] Kane AE, Mitchell SJ, Mach J, Huizer-Pajkos A, McKenzie C, Jones B, et al. Acetaminophen hepatotoxicity in mice: Effect of age, frailty and exposure type. Exp Gerontol 2016;73:95-106. doi: 10.1016/j.exger.2015.11.013.

[26] Jaeschke H, Ramachandran A. Oxidant stress and lipid peroxidation in acetaminophen hepatotoxicity. React Oxyg Species (Apex) 2018; 5(1):145-158.

[27] Ramos-Tovar E, Muriel P. Molecular mechanisms that link oxidative stress, inflammation, and fibrosis in the liver. Antioxidants 2020; 9(12):1279-1299. doi: 10.3390/antiox9121279.

[28] Vejerano EP, Rao G, Khachatryan L, Cormier SA, Lomnicki S. Environmentally persistent free radicals: Insights on a new class of pollutants. Environ Sci Technol 2018; 52(5):2468-2481. doi: 10.1021/acs.est.7b04439.

[29] Cheragh-Birjandi S, Moghbeli M, Haghighi F, Safdari MR, Baghernezhad M, Akhavan A, et al. Impact of resistance exercises and nano-curcumin on synovial levels of collagenase and nitric oxide in women with knee osteoarthritis. Trans Med Comm 2020; 5(1):3-8. doi: 10.1186/s41231-020-00055-0.

[30] Hassanzadeh K, Vahabzadeh Z, Bucarello L, Dragotto J, Corbo M, Maccarone R, et al. Protective effect of curcuma extract in an ex vivo model of retinal degeneration via antioxidant activity and targeting the SUMOylation. Oxid Med Cell Longev 2022;2022. doi: 10.1155/2022/8923615.

[31] Farhana A, Lappin SL. Biochemistry, lactate dehydrogenase. In: StatPearls. Treasure Island (FL): StatPearls Publishing; 2023.

[32] Wang Z, Wang X, Li X, Lu K, Wang L, Ma X, et al. Antioxidant effects of the aqueous extract of turmeric against hydrogen peroxide-induced oxidative stress in spotted seabass (Lateolabrax maculatus). Aquacult Fish 2024;9:71-77. doi: 10.1016/j.aaf.2022.11.003.

[33] Lopez-Castejon G, Brough D. Understanding the mechanism of IL-1β secretion. Cytokine Growth Factor Rev 2011; 22(4):189-195. doi: 10.1016/j.cytogfr.2011.10.001.

[34] Macías-Pérez JR, Aldaba-Muruato LR, Martínez-Hernández SL, Muñoz-Ortega MH, Pulido-Ortega J, Ventura-Juárez J. Curcumin provides hepatoprotection against amoebic liver abscess induced in hamster: Involvement of Nrf2/HO-1 and NF-κB/IL-1 signaling pathways. J Immunol Res 2019;2019. doi: 10.1155/2019/7431652.

[35] Liu Y, Liu Y, Huang X, Zhang J, Yang L. Protective effects and mechanism of curcumin on myocardial injury induced by coronary microembolization. J Cell Biochem 2019; 120(4):5695-5703. doi: 10.1002/jcb.27854.

[36] Rossi JF, Lu ZY, Jourdan M, Klein B. Interleukin-6 as a therapeutic target: Anti-IL6 therapy. Clin Cancer Res 2015; 21(6):1248-1257. doi: 10.1158/1078-0432.CCR-14-2291.

[37] Kang S, Tanaka T, Narazaki M, Kishimoto T. Targeting interleukin-6 signaling in clinic. Immunity 2019; 50(4):1007-1023. doi: 10.1016/j.immuni.2019.03.026.

[38] Vasanthkumar T, Hanumanthappa M, Lakshminarayana R. Curcumin and capsaicin modulate LPS-induced expression of COX-2, IL-6 and TGF-β in human peripheral blood mononuclear cells. Cytotechnology 2019; 71(5):963-976. doi: 10.1007/s10616-019-00338-x.

[39] Wu YR, Hsing CH, Chiu CJ, Huang HY, Hsu YH. Roles of IL-1 and IL-10 family cytokines in the progression of systemic lupus erythematosus: Friends or foes? IUBMB Life 2022;74:143-156. doi: 10.1002/iub.2568.

[40] Carlini V, Noonan DM, Abdalalem E, Goletti D, Sansone C, Calabrone L, et al. The multifaceted nature of IL-10: Regulation, role in immunological homeostasis and its relevance to cancer, COVID-19 and post-COVID conditions. Front Immunol 2023; 14. doi: 10.3389/fimmu.2023.1161067.

[41] Mardani R, Hamblin MR, Taghizadeh M, Banafshe HR, Nejati M, Mokhtari M, et al. Nanomicellar-curcumin exerts its therapeutic effects via affecting angiogenesis, apoptosis, and T cells in a mouse model of melanoma lung metastasis. Pathol Res Pract 2020; 216(9). doi: 10.1016/j.prp.2020.153082.

[42] Dhanasekaran DN, Reddy EP. JNK-signaling: A multiplexing hub in programmed cell death. Genes Cancer 2017;8:682-694. doi: 10.18632/genesandcancer.155.

[43] Jeong CW, Yoo KY, Lee SH, Jeong HJ, Lee CS, Kim SJ. Curcumin protects against regional myocardial ischemia/reperfusion injury through activation of RISK/GSK-3β and inhibition of p38 MAPK and JNK. J Cardiovasc Pharmacol Ther 2012; 17(4):387-394. doi: 10.1177/1074248412438102.

[44] Karamalakova YD, Nikolova GD, Georgiev TK, Gadjeva VG, Tolekova AN. Hepatoprotective properties of Curcuma longa L. extract in bleomycin-induced chronic hepatotoxicity. Drug Discov Ther 2019; 13(1):9-16. doi: 10.5582/ddt.2018.01081.

[45] Salehi E, Mashayekh M, Taheri F, Gholami M, Motaghinejad M, Safari S, et al. Curcumin as an effective agent for prevention or treatment of alcohol-induced toxicity in hepatocytes: An illustrated mechanistic review. Iran J Pharm Res 2021; 20(2):418-436. doi: 10.22037/ijpr.2020.112852.13985.