Genista cephalantha Spach. protects against acetaminophen-induced liver failure via preserving the glutathione redox system, reducing inflammatory response, and inhibiting hepatocyte death in rats

Document Type : Original Article


1 Laboratoire de Biologie et Environnement. Université Frères Mentouri Constantine 1, Algérie

2 Unité de Recherche, Valorisation des Ressources Naturelles, Molécules Bioactives et Analyses Physicochimiques et Biologiques. Frères Mentouri Constantine 1, Algérie

3 Research Laboratory Practice and Research Center, Igdir, University Igdir, Turkiye


Objective(s): The current study was conducted to assess the protective mechanisms of n-BuOH fraction from the aerial part of Genista cephontala (BEGC) on APAP-induced liver injury compared to necrostatine-1 (Nec-1).
Materials and Methods: A model of APAP-induced hepatotoxicity was created in male rats by injecting a single dose; 1000 mg/kg APAP, the protective effect was performed with (200 mg/kg; 10 days) BEGC compared to Nec-1, (1.8 mg/kg).
Results: BEGC or NeC-1 pretreatment significantly abolished impaired effects in APAP-rats, by decreasing the generation of TBARS and ROS in mitochondrial and cytosolic fractions and maintaining liver function activities. A marked response was observed in the levels of both GSH and GSH-system enzymes in liver homogenates and mitochondrial fractions to BEGC. BEGC/ Nec-1 successfully regulated the inflammatory mediators (IL-β, TNF-α, HMGB1, and acHMGB1) and MPO levels. During APAP treatment, no caspase-3 or -8 activity was detected, and the level of fk18; M30 was higher than the levels of cck18; M65. Moreover, RIPK3 and MLKL levels were increased in the APAP group. These results suggested that necroptosis predominates during the APAP liver injury model. Interestingly, these necroptotic factors were significantly down-regulated by BEGC treatment. Both biochemical and histopathological findings were consistent with each other.
Conclusion: From all these findings, the hepatoprotective effect of BEGC could be due to the abundance of polyphenols identified by LC-MS/MS analysis, as well as the synergistic interactions of all contents.


Main Subjects

1. Chiew AL, Gluud C, Brok J, Buckley NA. Interventions for paracetamol (acetaminophen) overdose. Cochrane Database Syst Rev 2018; 2:3328.
2. Yan HM, Ramachandran A, Bajt ML, Lemasters JJ, Jaeschke H. The oxygen tension modulates acetaminophen-induced mitochondrial oxidant stress and cell injury in cultured hepatocytes. Toxicol Sci 2010;117:515-523.
3. Malhi H, Gores GJ, Lemasters JJ. Apoptosis and necrosis in the liver: A tale of two deaths. Hepatology 2006; 43: 31-44.
4. Jaeschke H, Bajt M L. Intracellular signaling mechanisms of acetaminophen-induced liver cell death. Toxicol Sci 2006; 89: 31-41.
5. Galluzzi L, Kroemer G. Necroptosis: A specialized pathway of programmed necrosis. Cell 2008; 135: 1161-1163.
6. Yan M, Huo Y, Yin S, Hu H. Mechanisms of acetaminophen-induced liver injury and its implications for therapeutic interventions. Redox Biol 2018; 17: 274-283.
7. Cao L, Mu W. Necrostatin-1 and necroptosis inhibition: Pathophysiology and therapeutic implications. Pharmacol Res 2020; 163: 105297-105312.
8. Vanden BT, Vanlangenakker N, Parthoens E, Deckers W, Devos M, Festjens N, et al. Necroptosis, necrosis and secondary necrosis converge on similar cellular disintegration features. Cell Death Differ 2010; 17: 922-930.
9. Quezel P, Santa S. Nouvelle flore de l’Algérie et des régions désertiques méridionales. Tom I, Centre National de la Recherche Scientifique (CNRS): Paris, France, 1963; 470-471.
10. Chebbah K, Marchioni E, Menad A, Mekkiou R, Sarri D, Ameddah S, et al. Preliminary phytochemical screening, analysis of phenolic compounds and antioxidant activity of Genista cephalantha Spach. (Fabaceae). Inter J phytomedicine 2014; 6: 360-368. 
11. Kilkenny C, Browne WJ, Cuthill IC, Emerson M, Altman DG. Improving bioscience research reporting: The arrive guidelines for reporting animal research. J Pharmacol Pharmacother 2010; 1: 94-99.
12. McGill MR, Williams CD, Xie Y, Ramachandran A, Jaeschke H. Acetaminophen-induced liver injury in rats and mice: Comparison of protein adducts, mitochondrial dysfunction, and oxidative stress in the mechanism of toxicity. Toxicol Appl Pharmacol 2012; 264: 387-394.
13. Khodayar MJ, Kalantari H, Alidadi H, Khorsandi L, Ahangar N, Samimi A. Taurine attenuates valproic acid-induced hepatotoxicity via modulation of RIPK1/RIPK3/MLKL-mediated necroptosis signaling in mice. Mol Biol Rep 2021; 48: 4153-4162.
14.Johnson D, Lardy H. Isolation of liver or kidney mitochondria. Methods Enzymol 1967; 10: 94-96.
15. Gupta R, Dubey DK, Kannan GM, Flora SJ. Concomitant administration of Moringa oleifera seed powder in the remediation of arsenic-induced oxidative stress in mouse. Cell Biol Inter 2007; 31:44-56.
16. Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 1979; 95: 351-358.
17. Sedlak J, Hanus L. Changes of glutathione and protein bound SH-groups concentration in rat adrenals under acute and repeated stress. Endocrinol Exp 1982; 16 : 103-109.
18. Habig WH, Pabst MJ, Jakoby WB. Glutathione S-transferases the first enzymatic step in mercapturic acid formation. J Biol Chem 1974; 249: 7130-7139. 
19. Rotruck JT, Pope AL, Ganther HE, Swanson AB, Hafeman DG, Hoekstra WG. Selenium: Biochemical role as a component of glutathione peroxidase. Science 1973; 179: 588-590. 
20. Moss DW, Butterworth PJ. Enzymology and Medicine. London: Pitman Medical; p, 1974; 139.
21. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193:265-275. 
22. Jiang W, Reich III CF, Pisetsky DS. Mechanisms of activation of the RAW264.7 macrophage cell line by transfected mammalian DNA. Cell Immunol 2004;229:31-40.
23. Chen W, Bao G, Zhao L, Wang H. Analysis of circulating HMGB1 in Human Serum. Methods Mol Biol 2020:2108:15-28. 
24. Bradley PP, Priebat DA, Christenses RD, Rothstein G. Measurement of cutaneous inflammation: Estimation of neutrophil content with an enzyme marker. J Invest Dermatol 1982;78:206-209.
25. Ajiboye TO. Standardized extract of Vitex doniana sweet stalls protein oxidation, lipid peroxidation and DNA fragmention in acetaminophen induced hepatotoxicity. J Ethnopharmacol 2015; 164: 273-282.
26. Ajiboye TO. In vivo antioxidant potentials of Piliostigma thonningii (Schum) leaves: Studies on hepatic marker enzyme, antioxidant system, drug detoxifying enzyme and lipid peroxidation. Hum Exp Toxicol 2011; 30: 55-62.
27. Yuan L, Kaplowitz N. Mechanisms of drug-induced liver injury. Clin Liver Dis 2013; 17: 507-518.
28. Davis M, Ideo G, Harrison NG, Williams R. Hepatic glutathione depletion and impaired bromosulphthalein clearance early after paracetamol overdose in man and the rat. Clin Sci Mol Med 1975; 49: 495-502.
29. Ramachandran A, Lebofsky M, Baines CP, Lemasters JJ, Jaeschke H. Cyclophilin D deficiency protects against acetaminophen-induced oxidant stress and liver injury. Free Radic Res 2011; 45:156-164.
30. Abou Ghalia AH, Fouad IM. Glutathione and its metabolizing enzymes in patients with different benign and malignant diseases. Clin Biochem 2000; 33:657-662.
31. Hammond CL, Lee TK, Ballatori N. Novel roles for glutathione in gene expression, cell death, and membrane transport of organic solutes. J Hepatol 2001; 34: 946-954.
32. Sims NR, Nilsson M, Muyderman H. Mitochondrial glutathione: A modulator of brain cell death. J Bioenerg Biomembr 2004; 36: 329-333.
33. Arai M, Imai H, Koumura T, Yoshida M, Emoto K, Umeda M, et al. Mitochondrial phospholipid hydroperoxide glutathione peroxidase plays a major role in preventing oxidative injury to cells. J Biol Chem 1999; 274: 4924-4933.
34. Takemoto k, Hatano E, Iwaisako K, Takeiri M, Noma N, Ohmae S, et al. Necrostatin1protects against reactive oxygen species (ROS)-induced hepatotoxicity in acetaminophen-induced acute liver failure. FEBS Open Bio 2014; 4: 777-787.
35. Durgo K, Vukovic L, Rusak G, Osmak M, Colic JF. Effect of flavonoids in glutathione level, lipid peroxidation and cytochrome P450 CYP1A1 expression in human Laryngeal carcinoma cells. Food Technol Biotechnol 2007; 45: 69-79.
36. Jaeschke H, McGill MR, Ramachandran A. Oxidant stress, mitochondria, and cell death mechanisms in drug-induced liver injury: Lessons learned from acetaminophen hepatotoxicity. Drug Metab Rev 2012; 44: 88-106.
37. Jaeschke H, Duan L, Akakpo JY, Farhood A, Ramachandran A. The role of apoptosis in acetaminophen hepatotoxicity. Food Chem Toxicol 2018; 118: 709-718.
38. Gujral JS, Knight TR, Farhood A, Bajt ML, Jaeschke H. Mode of cell death after acetaminophen overdose in mice: Apoptosis or oncotic necrosis? Toxicol Sci 2002; 67: 322-328. 
39. Zhang YF, He W, Zhang C, Liu XJ, Lu Y, Wang H, et al. Role of receptor interacting protein (RIP)1 on apoptosis-inducing factor-mediated necroptosis during acetaminophen-evoked acute liver failure in mice. Toxicol Lett 2014; 225: 445-453.
40. Kramer G, Erdal H, Martens HJ, Nap M, Mauermann J, Steiner G, et al. Differentiation between cell death modes using measurements of different soluble forms of extracellular cytokeratin 18. Cancer Res 2004; 64: 1751-1756.
41. Antoine DJ, Williams DP, Kipar A, Jenkins RE, Regan SL, Sathish JG, et al. High-mobility group box-1 protein and keratin-18, circulating serum proteins informative of acetaminophen-induced necrosis and apoptosis in vivo. Toxicol Sci 2009; 112: 521-531. 
42. Jia Y, Feixia W, Qin G, Mengmeng L, Ling W, Zili Z, et al. Curcumol induces RIPK1/RIPK3 complex-dependent necroptosis via JNK1/2-ROS signaling in hepatic stellate cells. Redox Biol 2018; 19: 375-387.
43. Deutsch M, Graffeo CS, Rokosh R, Pansari M, Ochi A, Levie EM, et al. Divergent effects of RIP1 or RIP3 blockade in murine models of acute liver injury. Cell Death Dis. 2015; 6: 1759-1769.
44. Ramachandran A, McGill MR, Xie Y, Ni HM, Ding WX, Jaeschke H. Receptor interacting protein kinase 3 is a critical early mediator of acetaminophen-induced hepatocyte necrosis in mice. Hepatology 2013; 58: 2099-2108.
45. Pasparakis M, Vandenabeele P. Necroptosis and its role in inflammation. Nature 2015; 517: 311-320.
46. Lawson JA, Farhood A, Hopper RD, Bajt ML, Jaeschke H. The hepatic inflammatory response after acetaminophen overdose: Role of neutrophils. Toxicol Sci 2000; 54: 509-516.
47. Lau A, Wang S, Jiang J, Haig A, Pavlosky A, Linkermann A, et al. RIPK3-mediated necroptosis promotes donor kidney inflammatory injury and reduces allograft survival. Am J Transplant 2013; 13: 2805-2818.
48. Jaeschke H. Mechanisms of sterile inflammation in acetaminophen hepatotoxicity. Cell Mol Immunol 2018; 15: 74-75. 
49. Degterev A, Ofengeim D, Yuan J. Targeting RIPK1 for the treatment of human diseases. Proc Natl Acad Sci U S A 2019; 116: 9714-9722.
50. Xuan M, Okazaki M, Iwata N, Asano S, Kamiuchi S, Matsuzaki H, et al. Chronic treatment with a water-soluble extract from the culture medium of ganoderma lucidum mycelia prevents apoptosis and necroptosis in hypoxia/ischemia-induced injury of type 2 diabetic mouse brain. Evid Based Complement Alternat Med 2015; 865986-866001.
51. Dai MC, Zhonga ZH, Suna YH, Suna QF, Wang YT, Yanga GY, et al. Curcumin protects against iron induced neurotoxicity in primary cortical neurons by attenuating necroptosis. Neurotox Res 2013; 536: 41-46.
52. Baali N, Belloum Z, Baali S, Chabi B, Pessemesse L, Fouret G, et al. Protective activity of total polyphenols from Genista quadriflora Munby and Teucrium polium geyrii Maire in acetaminophen-induced hepatotoxicity in rats. Nutrients 2016; 8: 193-212. 
53. Ait-kaciAourahoun K, Fazouane F, Benayache S, Bettache Z, Benayad T ,Denni N. Antioxidant and anti-inflammatory activity of phenolic extracts of Genista ferox (Fabaceae) Pak. J Pharm Sci 2019; 32:2643-2649.
54. Abarikwu SO, Njoku RC, John IG, Amadi BA, Mgbudom-Okah CJ, Onuah CL. Antioxidant and anti-inflammatory protective effects of rutin and kolaviron against busulfan-induced testicular injuries in rats. Syst Biol Report Med 2022; 68: 151-161.
55. Skroza D, Simat V, Vrdolj L. Investigation of antioxidant synergisms and antagonisms among phenolic acids in the model matrices using FRAP and ORAC methods. Antioxidants 2022; 11: 1784-1798.