Carvacrol Exerts Anti‑Inflammatory, Anti‑Oxidative Stress and Hepatoprotective Effects Against Diclofenac‑Induced Liver Injury in Male Rats

Molood Barzan, Mahdi Heydari, Hamzeh Mirshekari‑ Jahangiri, Hassan Firouzi, Maryam Dastan, Mohammad Najaf, Mansoor Khaledi, Ali Nouri, Mehran Ebrahimi Shah-Abadi

Abstract


Background: Diclofenac (DIC) is an NSAID that can cause toxic effects in animals and humans and carvacrol (CAR) is a monoterpene compound that displays effective pharmacological and biological actions. The purpose of this work was to assess the influences of CAR on DIC‑induced liver injury and oxidative stress in male rats. Methods: The male Wistar rats were segregated into four groups. Group 1, the control group; Group 2 received DIC‑only (10 mg/kg BW, p.o); Group 3, received CAR‑only (10 mg/kg BW, p.o), and group 4 received DIC plus CAR. The serum levels as well as the activity of several liver‑associated markers, and oxidative and anti‑oxidant compounds were tested. The expression of pro‑inflammatory mediators was also studied using the qRT‑PCR analysis. Results: Our results showed that DIC treatment was associated with the elevation in the serum levels of liver‑related markers together with the increase in the serum and the hepatic levels of malondialdehyde (MDA) and protein carbonyl (PC). Moreover, DIC reduced the activity of the antioxidant system in the rats and increased lymphocyte infiltration into the hepatocytes. CAR; however, protected the hepatocytes from the toxic effects of DIC by enhancing the activity of antioxidant enzymes such as catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), and Glutathione (GSH). By diminishing the expression of tumor necrosis factor (TNF)‑α, CAR was also capable of preventing the inflammatory effects of DIC on liver cells. Conclusions: The findings of this study indicated that the administration of CAR could alleviate the noxious effects of DIC on the antioxidant defense system and liver tissue.

Keywords


Carvacrol; diclofenac; DIC‑induced liver injury; oxidative stress

Full Text:

PDF

References


Shitara Y, Sato H, Sugiyama Y. Evaluation of drug‑drug

interaction in the hepatobiliary and renal transport of drugs.

Annu Rev Pharmacol Toxicol 2005;45:689‑723.

Abdel‑Daim MM, Abdeen A. Protective effects of rosuvastatin

and vitamin E against fipronil‑mediated oxidative damage

and apoptosis in rat liver and kidney. Food Chem Toxicol

;114:69‑77.

Nair SS, Manalil JJ, Ramavarma SK, Suseela IM, Thekkepatt A,

Raghavamenon AC. Virgin coconut oil supplementation

ameliorates cyclophosphamide‑induced systemic toxicity in

mice. Hum Exp Toxicol 2016;35:205‑12.

Alabi QK, Akomolafe RO. Kolaviron diminishes

diclofenac‑induced liver and kidney toxicity in wistar rats via

suppressing inflammatory events, upregulating antioxidant

defenses, and improving hematological indices. Dose Response

;18:1559325819899256.

Harirforoosh S, West KO, Murrell DE, Denham JW, Panus PC,

Hanley GA. Examination of the pharmacodynamics and

pharmacokinetics of a diclofenac poly (lactic‑co‑glycolic) acid

nanoparticle formulation in the rat. Eur Rev Med Pharmacol Sci

;20:5021‑31.

Tomic Z, Milijasevic B, Sabo A, Dusan L, Jakovljevic V,

Mikov M, et al. Diclofenac and ketoprofen liver toxicity in rat.

Eur J Drug Metab Pharmacokinet 2008;33:253‑60.

Maity P, Bindu S, Choubey V, Alam A, Mitra K, Goyal M,

et al. Lansoprazole protects and heals gastric mucosa from

non‑steroidal anti‑inflammatory drug (NSAID)‑induced

gastropathy by inhibiting mitochondrial as well as Fas‑mediated

death pathways with concurrent induction of mucosal cell

renewal. J Biol Chem 2008;283:14391‑401.

Dhikav V, Singh S, Pande S, Chawla A, Anand KS.

Non‑steroidal drug‑induced gastrointestinal toxicity: Mechanisms

and management. J Indian Acad Clin Med 2003;4:315‑22.

Ahmed T, Archie SR, Faruk A, Chowdhury FA, Al Shoyaib A,

Ahsan CR. Evaluation of the anti‑inflammatory activities of

diclofenac sodium, prednisolone and atorvastatin in combination

with ascorbic acid. AntiInflamm AntiAllergy Agents Med Chem

;19:291‑301.

Aygün D, Kaplan S, Odaci E, Onger ME, Altunkaynak ME.

Toxicity of non‑steroidal anti‑inflammatory drugs: A review of

melatonin and diclofenac sodium association. Histol Histopathol

;27:417‑36.

Zhang Y, Geißen SU, Gal C. Carbamazepine and diclofenac:

Removal in wastewater treatment plants and occurrence in water

bodies. Chemosphere 2008;73:1151‑61.

Ahmed AY, Gad AM, El-Raouf OMA. Curcumin ameliorates

diclofenac sodium-induced nephrotoxicity in male albino rats.

J Biochem Mol Toxicol 2017;31:e21951. doi: 10.1002/jbt.

Simon JP, Evan Prince S. Aqueous leaves extract of Madhuca

longifolia attenuate diclofenac-induced hepatotoxicity: Impact on

oxidative stress, inflammation, and cytokines. J Cell Biochem

;119:6125‑35.

Esmaeilzadeh M, Heidarian E, Shaghaghi M, Roshanmehr H,

Najafi M, Moradi A, et al. Gallic acid mitigates

diclofenac‑induced liver toxicity by modulating oxidative stress

and suppressing IL‑1β gene expression in male rats. Pharm Biol

;58:590‑6.

Suntres ZE, Coccimiglio J, Alipour M. The bioactivity and

toxicological actions of carvacrol. Crit Rev Food Sci Nutr

;55:304‑18.

Hotta M, Nakata R, Katsukawa M, Hori K, Takahashi S, Inoue H.

Carvacrol, a component of thyme oil, activates PPARα and γ and

suppresses COX‑2 expression [S]. J Lipid Res 2010;51:132‑9.

Guimarães AG, Xavier MA, de Santana MT, Camargo EA,

Santos CA, Brito FA, et al. Carvacrol attenuates mechanical

hypernociception and inflammatory response. Naunyn

Schmiedebergs Arch Pharmacol 2012;385:253‑63.

Uyanoglu M, Canbek M, Ceyhan E, Senturk H, Bayramoglu G,

Gunduz O, et al. Preventing organ injury with carvacrol after

renal ischemia/reperfusion. J Med Plants Res 2011;5:72‑80.

Ahmadvand H, Tavafi M, Asadollahi V, Jafaripour L,

Hadipour‑Moradi F, Mohammadrezaei‑Khoramabadi R,

et al. Protective effect of carvacrol on renal functional and

histopathological changes in gentamicin‑induced‑nephrotoxicity

in rats. Zahedan J Res Med Sci 2016;18:e6446.

Bozkurt M, Em S, Oktayoglu P, Turkcu G, Yuksel H,

Sarıyıldız MA, et al. Carvacrol prevents methotrexate‑induced

renal oxidative injury and renal damage in rats. Clin Invest Med

;37:E19‑25.

Gunes S, Ayhanci A, Sahinturk V, Altay DU, Uyar R. Carvacrol

attenuates cyclophosphamide‑induced oxidative stress in rat

kidney. Can J Physiol Pharmacol 2017;95:844‑9.

Motawi TK, Ahmed SA, El‑Boghdady NA, Metwally NS,

Nasr NN. Impact of betanin against paracetamol and diclofenac

induced hepato‑renal damage in rats. Biomarkers 2020;25:86‑93.

Hagan EC, Hansen WH, Fitzhugh OG, Jenner PM, Jones WI,

Taylor JM, et al. Food flavourings and compounds of related

structure. II. Subacute and chronic toxicity. Food Cosmet Toxicol

;5:141‑57.

Nouri A, Hashemzadeh F, Soltani A, Saghaei E, Amini‑Khoei H.

Progesterone exerts antidepressant‑like effect in a mouse

model of maternal separation stress through mitigation of

neuroinflammatory response and oxidative stress. Pharm Biol

;58:64‑71.

Nouri A, Heidarian E, Amini‑Khoei H, Abbaszadeh S, Basati G.

Quercetin through mitigation of inflammatory response and

oxidative stress exerts protective effects in rat model of

diclofenac‑induced liver toxicity. J Pharm Pharmacogn Res

;7:200‑12.

Ellman GL. Tissue sulfhydryl groups. Arch Biochem Biophys

;82:70‑7.

Flohe L, Otting F. Superoxide dismutase assays. Methods

Enzymol 1984;105:93‑104.

Bradford MM. A rapid and sensitive method for the quantitation

of microgram quantities of protein utilizing the principle of

protein‑dye binding. Anal Biochem 1976;72:248‑54.

Aebi H. Catalase. Methods of Enzymatic Analysis. Elsevier;

p. 673‑84.

Reznick AZ, Packer L. Oxidative damage to proteins:

Spectrophotometric method for carbonyl assay. Methods

Enzymol 1994;233:357‑63.

Livak KJ, Schmittgen TD. Analysis of relative gene expression

data using real‑time quantitative PCR and the 2− ΔΔCT method.

Methods 2001;25:402‑8.

Adeyemi WJ, Olayaki LA. Diclofenac–induced hepatotoxicity: Low dose of omega‑3 fatty acids have more protective effects.

Toxicol Rep 2017;5:90‑5.

Alabi QK, Akomolafe RO, Olukiran OS, Adeyemi WJ,

Nafiu AO, Adefisayo MA, et al. The Garcinia kola biflavonoid

kolaviron attenuates experimental hepatotoxicity induced by

diclofenac. Pathophysiology 2017;24:281‑90.

Mohseni R, Karimi J, Tavilani H, Khodadadi I, Hashemnia M.

Carvacrol ameliorates the progression of liver fibrosis

through targeting of Hippo and TGF‑β signaling pathways

in carbon tetrachloride (CCl4)‑induced liver fibrosis in rats.

Immunopharmacol immunotoxicol 2019;41:163‑71.

Aristatile B, Al-Numair KS, Veeramani C, Pugalendi KV. Effect

of carvacrol on hepatic marker enzymes and antioxidant status

in d-galactosamine-induced hepatotoxicity in rats. Fundam Clin

Pharmacol 2009;23:757‑65.

Bozkurt M, Bodakci MN, Turkcu G, Kuyumcu M, Akkurt M,

Sula B, et al. Protective effects of carvacrol against

methotrexate‑induced liver toxicity in rats. Acta Chir Belg

;114:404‑9.

Gaweł S, Wardas M, Niedworok E, Wardas P.

Malondialdehyde (MDA) as a lipid peroxidation marker. Wiad

Lek 2004;57:453‑5.

Heidarian E, Nouri A. Hepatoprotective effects of silymarin

against diclofenac‑induced liver toxicity in male rats based on

biochemical parameters and histological study. Arch Physiol

Biochem 2019;127:1‑7.

Ogbe RJ, Luka CD, Adoga GI. Effect of aqueous ethanol extract

of Dialium guineense leaf on diclofenac‑induced oxidative

stress and hepatorenal injuries in Wistar rats Comp Clin Path

;28:241‑8.

Safari T, Miri S, Gharaei FK, NazriPanjaki A, Saeidienik F,

Bagheri H, et al. Nitric oxide metabolite changes in

gentamicin‑induced nephrotoxicity; The effects of antioxidant

vitamins. J Renal inj Prev 2018;7:201‑5.

Wei T, Tian W, Liu F, Xie G. Protective effects of exogenous

β‑hydroxybutyrate on paraquat toxicity in rat kidney. Biochem

Biophys Res Commun 2014;447:666‑71.

Giridharan R, Lavinya U, Sabina EP. Suppressive effect of

Spirulina fusiformis on diclofenac‑induced hepato‑renal injury

and gastrointestinal ulcer in Wistar albino rats: A biochemical

and histological approach. Biomed Pharmacother 2017;88:11‑8.

Ramezannezhad P, Nouri A, Heidarian E. Silymarin mitigates

diclofenac‑induced liver toxicity through inhibition of

inflammation and oxidative stress in male rats. J Herbmed

Pharmacol 2019;8:231‑7.

Samarghandian S, Farkhondeh T, Samini F, Borji A. Protective

effects of carvacrol against oxidative stress induced by chronic

stress in rat’s brain, liver, and kidney. Biochem Res Int

;2016:2645237.

Nouri A, Heidarian E, Nikoukar M. Effects of N‑acetyl

cysteine on oxidative stress and TNF‑α gene expression in

diclofenac‑induced hepatotoxicity in rats. Toxicol Mech Methods

;27:561‑7.

Ahmad I, Shukla S, Kumar A, Singh BK, Kumar V,

Chauhan AK, et al. Biochemical and molecular mechanisms

of N‑acetyl cysteine and silymarin‑mediated protection against

maneb‑and paraquat‑induced hepatotoxicity in rats. Chem‑Biol

Interact 2013;201:9‑18.

Niu X, de Graaf IA, van de Vegte D, Langelaar‑Makkinje M,

Sekine S, Groothuis GM. Consequences of Mrp2 deficiency for

diclofenac toxicity in the rat intestine ex vivo. Toxicol In Vitro

;29:168‑75.

Nouri A, Heidarian E. Ameliorative effects of N-acetyl

cysteine on diclofenac-induced renal injury in male rats based

on serum biochemical parameters, oxidative biomarkers, and

histopathological study. J Food Biochem 2019;43:e12950.

Sharifi‑Rigi A, Heidarian E, Amini SA. Protectie and

anti‑inflammatory effects of hydroalcoholic leaf extract of

Origanum vulgare on oxidative stress, TNF‑α gene expression

and liver histological changes in paraquat‑induced hepatotoxicity

in rats. Arch Physiol Biochem 2019;125:56‑63.

Laskin DL, Laskin JD. Role of macrophages and inflammatory

mediators in chemically induced toxicity. Toxicology

;160:111‑8.

Pang L, Ye W, Che XM, Roessler BJ, Betz AL, Yang GY.

Reduction of inflammatory response in the mouse brain with

adenoviral‑mediated transforming growth factor‑β1 expression.

Stroke 2001;32:544‑52.