Lagerstroemia indica methanolic extract alleviates CCl₄-induced liver toxicity via modulation of Nrf2 signaling pathway in rats

Abstract

Objective: To assess the hepatoprotective effects of methanolic extract of Lagerstroemia indica leaves on carbon tetrachloride (CCl4)-induced liver damage in rats.

Methods: Fifty male Sprague Dawley rats were divided into five groups: the normal control, the CCl4 group, the CCl4 group treated with silymarin (200 mg/kg) or Lagerstroemia indica extract (200 and 400 mg/kg). Oxidative stress, inflammatory, and apoptotic markers, as well as liver enzymes were analyzed. Histopathological studies were performed to examine the effect of the plant extract on liver structure. In addition, quantitative RT-PCR analysis was carried out to determine Nrf2, Keapl, HMOX1, and NQO1 expression.

Results: Lagerstroemia indica extract significantly improved liver function by reducing aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, and bilirubin levels. It also enhanced the activities of catalase, superoxide dismutase, and glutathione peroxidase, increased glutathione content, and reduced the levels of malondialdehyde, nuclear factor kappa-B, tumor necrosis factor-α, interleukin-1β, interleukin-6, cyclooxygenase-2, Bax, caspase-3 and caspase-9. In addition, the mRNA expression levels of Nrf2, Keapl, HMOX1, and NQO1 were markedly downregulated by treatment with Lagerstroemia indica extracts. Histopathological examination revealed the extract confered protection against CCl4-induced liver damage as evidenced by reduced sinusoidal dilatation, thinner fibrotic septa, decreased vacuolar degeneration, and improved hepatic cord organization.

Conclusions: Lagerstroemia indica extract possesses hepatoprotective activity by modulating the Nrf2-Keap1-HMOX1-NQO1 signaling pathway in a rat model of CCl4-induced liver damage.

Keywords

Lagerstroemia indica / CCl₄ / Liver disease / Nrf2 / Oxidative stress

Conflict of interest statement

The authors declare that there is no conflict of interest.

Funding

The study received no extramural funding.

Data availability statement

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

Authors’ contributions

JN, SARS, HF and IM contributed to the conception or design of the work. JN, SY, and HF performed data collection. MS, SARS, SY, HM, and IM carried out data analysis and interpretation. JN, HM, HF, and IM drafted the article. MS, SARS, SY and IM made critical revision of the article. All authors made final approval of the version to be published.

Publisher’s note

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

References

[1] Meech R, Hu DG, McKinnon RA, Mubarokah SN, Haines AZ, Nair PC, et al. The UDP-glycosyltransferase (UGT) superfamily: New members, new functions, and novel paradigms. Physiol Rev 2019; 99(2):1153-1222.

[2] Moreno-Torres M, Quintás G, Castell JV. The potential role of metabolomics in drug-induced liver injury (DILI) assessment. Metabolites 2022; 12(6):564.

[3] Bencheikh N, Bouhrim M, Kharchoufa L, Choukri M, Bnouham M, Elachouri M. Desf.) Protective effect of Zizyphus lotus L. ( fruit against CCl₄-induced acute liver injury in rat. Evid Based Complement Alternat Med 2019;2019. doi: 10.1155/2019/6161593.

[4] Neuman MG. Hepatotoxicity:Mechanisms of liver injury. In: Radu-Ionita F, Pyrsopoulos N, Jinga M, Tintoiu I, Sun Z, Bontas E (eds). Liver diseases: A multidisciplinary textbook. Springer, Cham; 2020, p. 75-84.

[5] Zodape G, Bhise P. Effect of Aloe vera extract and isoniazid-rifampicin drug on liver histological studies of male Wistar rats. Int J Pharm Sci Res 2018; 9(10):4318-4325.

[6] Kruk J, Aboul-Enein BH, Duchnik E, Marchlewicz M. Antioxidative properties of phenolic compounds and their effect on oxidative stress induced by severe physical exercise. J Physiol Sci 2022; 72(1):1-24.

[7] Adachi T, Nakamura Y. Aptamers: A review of their chemical properties and modifications for therapeutic application. Molecules 2019; 24(23):4229.

[8] Hussain F, Pathan S, Sahu K, Gupta BK. Herbs as cosmetics for natural care: A review. GSC Biol Pharmacol Sci 2022; 19(2):316-322.

[9] Yang EJ, Lee JS, Song BB, Yun CY, Kim DH, Kim IS. Anti-inflammatory effects of ethanolic extract from Lagerstroemia indica on airway inflammation in mice. J Ethnopharmacol 2011; 136(3):422-427.

[10] Al-Snafi AE. A review on Lagerstroemia indica: A potential medicinal plant. IOSR J Pharm 2019; 9(6):36-42.

[11] Nisar J, Ali Shah SM, Ayaz S, Akram M, Rashid A, Mustafa I, et al. In vitro comparative evaluation of Tamarix gallica extracts for antioxidant and antidiabetic activity. Exp Biol Med 2023; 248(3):253-262.

[12] Mustafa I, Faisal MN, Hussain G, Muzaffar H, Imran M, Ijaz MU, et al. Efficacy of Euphorbia helioscopia in context to a possible connection between antioxidant and antidiabetic activities: A comparative study of different extracts. BMC Complement Med Ther 2021; 21(1):1-12.

[13] Boota M, Shah SMA, Rashid A, Akram M, Ayaz S, Mustafa I, et al. The hepatoprotective and anti-nephrotoxic potential of methanolic extract of a polyherbal preparation in CCl₄-induced liver injury model of wistar rats. Dose Response 2022; 20(3). doi: 10.1177/15593258221124728.

[14] Chang M, Ahmed AF, Cui L. and The hypoglycemic effect of Lagerstroemia indica L. Lagerstroemia indica L. f. alba (Nichols.) Rehd. in vitro and in vivo. J Future Food 2023; 3(3):273-277.

[15] Mustafa I, Anwar H, Irfan S, Muzaffar H, Ijaz MU. Attenuation of carbohydrate metabolism and lipid profile by methanolic extract of Euphorbia helioscopia and improvement of beta cell function in a type 2 diabetic rat model. BMC Complement Med Ther 2022; 22(1):23.

[16] Lawrence RA, Burk RF. Glutathione peroxidase activity in selenium-deficient rat liver. Biochem Biophys Res Commun 1976; 71(4):952-958.

[17] Sun Y, Oberley LW, Li Y. A simple method for clinical assay of superoxide dismutase. Clin Chem 1988; 34(3):497-500.

[18] Jollow D, Mitchell J, Zampaglione N, Gillette J. Bromobenzene-induced liver necrosis. Protective role of glutathione and evidence for 3, 4-bromobenzene oxide as the hepatotoxic metabolite. Pharmacology 1974; 11(3):151-169.

[19] Sun T, Wang BY, Li XY, Zhang XZ, Lei YF, Yu SF, et al. Apigenin ameliorates CCl₄-induced hepatic fibrosis by inhibiting hepatic stellate cell activation via PI3K/AKT/GSK3β pathway in mice. Asian Pac J Trop Biomed 2025; 15(4):141-149.

[20] Leaker BD, Wang Y, Tam J, Anderson RR. Analysis of culture and RNA isolation methods for precision-cut liver slices from cirrhotic rats. Sci Rep 2024; 14(1):15349.

[21] Presti G, Guarrasi V, Gulotta E, Provenzano F, Provenzano A, Giuliano S, et al. Bioactive compounds from extra virgin olive oils: Correlation between phenolic content and oxidative stress cell protection. Biophys Chem 2017; 230:109-116.

[22] Ajaib M, Arooj T, Khan KM, Farid S, Ishtiaq M, Perveen S, et al. Phytochemical, antimicrobial and antioxidant screening of fruits, bark and leaves of Lagerstroemia indica. J Chem Society Pak 2016; 38(3).

[23] Iqbal N, Zubair HM, Almutairi MH, Abbas M, Akhtar MF, Aleya L, et al. Hepatoprotective effect of Cordia rothii extract against CCl₄-induced oxidative stress via Nrf2-NFκB pathways. Biomed Pharmacother 2022; 156:113840.

[24] Ma P, Xiao H, Yu C, Liu J, Cheng Z, Song H, et al. Enhanced cisplatin chemotherapy by iron oxide nanocarrier-mediated generation of highly toxic reactive oxygen species. Nano Lett 2017; 17(2):928-937.

[25] Yang J, Kong XD, Zhu-Salzman K, Qin QM, Cai QN. The key glutathione S-transferase family genes involved in the detoxification of rice gramine in brown planthopper Nilaparvata lugens. Insects 2021; 12(12):1055.

[26] Tang SP, Mao XI, Chen YH, Yan LI, Ye LP, Li SW. Reactive oxygen species induce fatty liver and ischemia-reperfusion injury by promoting inflammation and cell death. Front Immunol 2022; 13:870239.

[27] Chandana L, Bhikshapathi D. Free radical scavenging activity of Pleurotus ostreatus against CCl₄-induced hepatic damage in Wister rats. Int J Pharm Pharm Sci 2023:17-22.

[28] Akbari B, Najafi F, Bahmaei M, Mahmoodi NM, Sherman JH. Modeling and optimization of malondialdehyde (MDA) absorbance behavior through response surface methodology (RSM) and artificial intelligence network (AIN): An endeavor to estimate lipid peroxidation by determination of MDA. J Chemomet 2023; 37(4):e3468.

[29] Procházková D, Boušová I, Wilhelmová N. Antioxidant and prooxidant properties of flavonoids. Fitoterapia 2011; 82(4):513-523.

[30] Estevao C, Bowers CE, Luo D, Sarker M, Hoeh AE, Frudd K, et al. CCl₄ induces inflammatory signalling and barrier disruption in the neurovascular endothelium. Brain Behav Immun Health 2021; 18:100370.

[31] Liu J, Han X, Zhang T, Tian K, Li Z, Luo F. Reactive oxygen species (ROS) scavenging biomaterials for anti-inflammatory diseases: From mechanism to therapy. J Hematol Oncol 2023; 16(1):116.

[32] Hariharan A, Hakeem AR, Radhakrishnan S, Reddy MS, Rela M. The role and therapeutic potential of NF-kappa-B pathway in severe COVID-19 patients. Inflammopharmacology 2021; 29:91-100.

[33] Cui J, Jia J. Natural COX-2 inhibitors as promising anti-inflammatory agents: An update. Curr Med Chem 2021; 28(18):3622-3646.

[34] Kostic T, Popović D, Perisic Z, Stanojevic D, Dakic S, Saric S, et al. The hepatoprotective effect of aminoguanidine in acute liver injury caused by CCl₄ in rats. Biomed Pharmacother 2022; 156:113918.

[35] Wolf P, Schoeniger A, Edlich F. Pro-apoptotic complexes of BAX and BAK on the outer mitochondrial membrane. Biochim Biophys Acta Mol Cell Res 2022; 1869(10):119317.

[36] Yang CC, Chen YT, Chen CH, Chiang JY, Zhen YY, Yip HK. Assessment of doxorubicin-induced mouse testicular damage by the novel second-harmonic generation microscopy. Am J Transl Res 2017; 9(12):5275-5288.

[37] Chipuk JE, Green DR. PUMA cooperates with direct activator proteins to promote mitochondrial outer membrane permeabilization and apoptosis. Cell Cycle 2009; 8(17):2692-2696.

[38] Al-Megrin WA, Alkhuriji AF, Yousef AOS, Metwally DM, Habotta OA, Kassab RB, et al. Antagonistic efficacy of luteolin against lead acetate exposure-associated with hepatotoxicity is mediated via antioxidant, anti-inflammatory, and anti-apoptotic activities. Antioxidants 2019; 9(1):10.

[39] Othman MS, Khaled AM, Al-Bagawi AH, Fareid MA, Ghany RA, Habotta OA, et al. Hepatorenal protective efficacy of flavonoids from Ocimum basilicum extract in diabetic albino rats: A focus on hypoglycemic, antioxidant, anti-inflammatory and anti-apoptotic activities. Biomed Pharmacother 2021; 144:112287.

[40] Magyar K, Szende B. (-)-Deprenyl, a selective MAO-B inhibitor, with apoptotic and anti-apoptotic properties. Neurotoxicology 2004; 25(1-2):233-242.

[41] Jayashree G, Kumar KH, Krupashree K, Rachitha P, Khanum F. LC-ESI-MS/MS analysis of Asparagus racemosus Willd. roots and its protective effects against t-BHP induced oxidative stress in rats. Ind Crop Prod 2015; 78:102-109.

[42] Farhan AS. Alcohol extract of Tribulus terrestris ameliorates liver of mice exposed to cadmium acetate. JUAPS 2019; 13(2):10-15.

[43] Okorie N, Fasogbon S, Ajileye A, Adegeye F. Effects of Mucuna pruriens leaf extract on the liver of wister rats. J Med Lab Sci 2020; 30(1):107-117.

[44] Louisa M, Putera AM, Sidqi AA, Mahaputra DK, Firmansyah ER, Ammar MF, et al. Attenuation of cisplatin-induced hepatotoxicity by nanocurcumin through modulation of antioxidative and anti-inflammatory pathways. J App Pharm Sci 2023; 13(3):60-70.

[45] Ulasov AV, Rosenkranz AA, Georgiev GP, Sobolev AS. Nrf2/Keap1/ARE signaling: Towards specific regulation. Life Sci 2022; 291:120111.

[46] Somade OT, Ajiboye BO, Osukoya OA, Jarikre TA, Oyinloye BE. Syringic acid ameliorates testicular oxidative stress via the conservation of endogenous antioxidant markers and inhibition of the activated Nrf2-Keap1-NQO1-HO1 signaling in methyl cellosolve-administered rats. Pharmacol Res Mod Chin Med 2023; 6:100207.