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BMC Complementary Medicine and Therapies

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Open Access 01-12-2015 | Research article

Hepatoprotective effects of Micromeria croatica ethanolic extract against CCl₄–induced liver injury in mice

Authors: Sanda Vladimir-Knežević, Olga Cvijanović, Biljana Blažeković, Marija Kindl, Maja Bival Štefan, Robert Domitrović

Published in: BMC Complementary Medicine and Therapies | Issue 1/2015

Abstract

Background

Micromeria croatica (Pers.) Schott is an aromatic plant from Lamiaceae family previously found to possess potent in vitro antioxidant activity which is mainly attributed to the high level of polyphenolic substances. The aim of this study was to investigate the hepatoprotective activity and possible underlying mechanisms of Micromeria croatica ethanolic extract (MC) using a model of carbon tetrachloride (CCl₄)-induced liver injury in mice.

Methods

Male BALB/cN mice were randomly divided into seven groups: control group received saline, MC group received ethanolic extract of M. croatica in 5 % DMSO (100 mg/kg b.w., p.o.), and CCl₄ group was administered CCl₄ dissolved in corn oil (2 mL/kg, 10 % v/v, ip). MC50, MC200 and MC400 groups were treated with MC orally at doses of 50, 200 and 400 mg/kg once daily for 2 consecutive days, respectively, 6 h after CCl₄ intoxication. The reference group received silymarin at dose of 400 mg/kg. At the end of experiment, blood and liver samples were collected for biochemical, histopathological, immunohistochemical and Western blot analyses. In addition, major phenolic compounds in MC were quantified by HPLC-DAD.

Results

CCl₄ intoxication resulted in liver cells damage and oxidative stress and triggered inflammatory response in mice livers. MC treatment decreased ALT activity and prevented liver necrosis. Improved hepatic antioxidant status was evident by increased Cu/Zn SOD activity and decreased 4-hydroxynonenal (4-HNE) formation in the livers. Concomitantly, nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) were overexpressed. The hepatoprotective activity of MC was accompanied by the increase in nuclear factor-kappaB (NF-κB) activation and tumor necrosis factor-alpha (TNF-α) expression, indicating amelioration of hepatic inflammation. Additionally, MC prevented tumor growth factor-β1 (TGF-β1) and α-smooth muscle actin (α-SMA) expression, suggesting the potential for suppression of hepatic fibrogenesis.

Conclusion

These results of the present study demonstrated that MC possesses in vivo antioxidant and anti-inflammatory activity and exhibits antifibrotic potential, which are comparable to those of standard hepatoprotective compound silymarin.

Blachier M, Leleu H, Peck-Radosavljevic M, Valla DC, Roudot-Thoraval F. The burden of liver disease in Europe: a review of available epidemiological data. J Hepatol. 2013;58:593–608.CrossRefPubMed

Zhu R, Wang Y, Zhang L, Guo Q. Oxidative stress and liver disease. Hepatol Res. 2012;42:741–9.CrossRefPubMed

Stickel F, Schuppan D. Herbal medicine in the treatment of liver diseases. Dig Liver Dis. 2007;39:293–304.CrossRefPubMed

Amat N, Upur H, Blažeković B. In vivo hepatoprotective activity of the aqueous extract of Artemisia absinthium L. against chemically and immunologically induced liver injuries in mice. J Ethnopharmacol. 2010;131:478–84.CrossRefPubMed

Kamisan FH, Yahya F, Mamat SS, Fahmi Kamarolzaman MF, Mohtarrudin N, Kek TL, et al. Effect of methanol extract of Dicranopteris linearis against carbon tetrachloride- induced acute liver injury in rats. BMC Compl Alter Med. 2014;14:123.CrossRef

Bräuchler C, Meimberg H, Abele T, Heubl G. Polyphyly of the genus Micromeria (Lamiaceae): Evidence from cpDNA sequence data. Taxon. 2005;54:639–50.CrossRef

Öztürk M, Kolak U, Topçu G, Öksüz S, Choudhary MI. Antioxidant and anticholinesterase active constituents from Micromeria cilicica by radical-scavenging activity-guided fractionation. Food Chem. 2011;126:31–8.CrossRef

Redžić S. Wild medicinal plants and their usage in traditional human therapy (Southern Bosnia and Herzegovina, W. Balkan). J Med Plant Res. 2010;4:1003–27.

Said O, Khalil K, Fulder S, Azaizeh H. Ethnopharmacological survey of medicinal herbs in Israel, the Golan Heights and the West Bank region. J Ethnopharmacol. 2002;83:251–65.CrossRefPubMed

10.

Vladimir-Knežević S, Blažeković B, Bival Štefan M, Alegro A, Kőszegi T, Petrik J. Antioxidant activities and polyphenolic contents of three selected Micromeria species from Croatia. Molecules. 2011;16:1454–70.CrossRefPubMed

11.

Herken EN, Celik A, Aslan M, Aydınlık N. The constituents of essential oil: antimicrobial and antioxidant activity of Micromeria congesta Boiss. & Hausskn. ex Boiss. from East Anatolia. J Med Food. 2012;15:835–9.CrossRefPubMed

12.

Lee IT, Luo SF, Lee CW, Wang SW, Lin CC, Chang CC, et al. Overexpression of HO-1 protects against TNF-α-mediated airway inflammation by down-regulation of TNFR1-dependent oxidative stress. Am J Pathol. 2009;175:519–32.CrossRefPubMedPubMedCentral

13.

Shehab NG, Abu-Gharbieh E. Constituents and biological activity of the essential oil and the aqueous extract of Micromeria fruticosa (L.) Druce subsp. serpyllifolia. Pak J Pharm Sci. 2012;25:687–92.PubMed

14.

Šilić Č. Endemične biljke. Svjetlost: Sarajevo; 1984.

15.

Tomas-Barberan FA, Gil MI, Marin PD, Tomas-Lorente F. Flavonoids from some Yugoslavian Micromeria species: Chemotaxnomical aspects. Biochem Syst Ecol. 1991;19:697–8.CrossRef

16.

Stanić G, Kalođera Z, Petričić J, Todorić A, Blažević N. Essential oil content and composition of Micromeria croatica (Pers.) Schott and Micromeria thymifolia (Scop.) Fritsch. Acta Pharm Jugosl. 1988;38:251–4.

17.

Fecka I, Turek S. Determination of water-soluble polyphenolic compounds in commercial herbal teas from Lamiaceae: peppermint, melissa, and sage. J Agric Food Chem. 2007;55:10908–17.CrossRefPubMed

18.

Domitrović R, Škoda M, Vasiljev Marchesi V, Cvijanović O, Pernjak Pugel E, Bival Štefan M. Rosmarinic acid ameliorates acute liver damage and fibrogenesis in carbon tetrachloride-intoxicated mice. Food Chem Toxicol. 2013;51:370–8.CrossRefPubMed

19.

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.CrossRefPubMed

20.

Domitrović R, Jakovac H, Vasiljev Marchesi V, Šain I, Romić Ž, Rahelić D. Preventive and therapeutic effects of oleuropein against carbon tetrachloride-induced liver damage in mice. Pharmacol Res. 2012;65:451–64.CrossRefPubMed

21.

Sun F, Hamagawa E, Tsutsui C, Ono Y, Ogiri Y, Kojo S. Evaluation of oxidative stress during apoptosis and necrosis caused by carbon tetrachloride in rat liver. BBA-Mol Basis Dis. 2001;1535:186–91.CrossRef

22.

Upur H, Amat N, Blažeković B, Talip A. Protective effect of Cichorium glandulosum root extract on carbon tetrachloride-induced and galactosamine-induced hepatotoxicity in mice. Food Chem Toxicol. 2009;47:2022–30.CrossRefPubMed

23.

Maheshwari DT, Yogendra Kumar MS, Verma SK, Singh VK, Singh SN. Antioxidant and hepatoprotective activities of phenolic rich fraction of Seabuckthorn (Hippophae rhamnoides L.) leaves. Food Chem Toxicol. 2011;49:2422–8.CrossRefPubMed

24.

Poli G, Biasi F, Leonarduzzi G. 4-Hydroxynonenal-protein adducts: A reliable biomarker of lipid oxidation in liver diseases. Mol Asp Med. 2008;29:67–71.CrossRef

25.

Paine A, Eiz-Vesper B, Blasczyk R, Immenschuh S. Signaling to heme oxygenase-1 and its anti-inflammatory therapeutic potential. Biochem Pharmacol. 2010;80:1895–903.CrossRefPubMed

26.

Kumar KJS, Chu FH, Hsieh HW, Liao JW, Li WH, Lin JCC, et al. Antroquinonol from ethanolic extract of mycelium of Antrodia cinnamomea protects hepatic cells from ethanol-induced oxidative stress through Nrf-2 activation. J Ethnopharmacol. 2011;136:168–77.CrossRefPubMed

27.

Abraham NG, Kappas A. Pharmacological and clinical aspects of heme oxygenase. Pharmacol Rev. 2008;60:79–127.CrossRefPubMed

28.

Wu YH, Hao BJ, Cao HC, Xu W, Li YJ, Li LJ: Anti-hepatitis B virus effect and possible mechanism of action of 3,4-o-dicaffeoylquinic ycid in vitro and in vivo. Evid Based Complement Alternat Med. 2012;2012:356806.

29.

Liu S, Hou W, Yao P, Li N, Zhang B, Hao L, et al. Heme oxygenase-1 mediates the protective role of quercetin against ethanol-induced rat hepatocytes oxidative damage. Toxicol In Vitro. 2012;26:74–80.CrossRefPubMed

30.

Park EJ, Kim YM, Park SW, Kim HJ, Lee JH, Lee DU, et al. Induction of HO-1 through p38 MAPK/Nrf2 signaling pathway by ethanol extract of Inula helenium L. reduces inflammation in LPS-activated RAW 264.7 cells and CLP-induced septic mice. Food Chem Toxicol. 2013;55:386–95.CrossRefPubMed

31.

Tak PP, Firestein GS. NF-κB: a key role in inflammatory diseases. J Clin Invest. 2001;107:7–11.CrossRefPubMedPubMedCentral

32.

Muriel P. NF-kappaB in liver diseases: a target for drug therapy. J Appl Toxicol. 2009;29:91–100.CrossRefPubMed

33.

Luedde T, Schwabe RF. NF-κB in the liver-linking injury, fibrosis and hepatocellular carcinoma. Nat Rev Gastroenterol Hepatol. 2011;8:108–18.CrossRefPubMedPubMedCentral

34.

Weber LW, Boll M, Stampfl A. Hepatotoxicity and mechanism of action of haloalkanes: carbon tetrachloride as a toxicological model. Crit Rev Toxicol. 2003;33:105–36.CrossRefPubMed

35.

Domitrović R, Jakovac H. Antifibrotic activity of anthocyanidin delphinidin in carbon tetrachloride-induced hepatotoxicity in mice. Toxicology. 2010;272:1–10.CrossRefPubMed

36.

Parola M, Robino G. Oxidative stress-related molecules and liver fibrosis. J Hepatol. 2001;35:297–306.CrossRefPubMed

37.

Liu JY, Chen CC, Wang WH, Hsu JD, Yang MY, Wang CJ. The protective effects of Hibiscus sabdariffa extract on CCl₄-induced liver fibrosis in rats. Food Chem Toxicol. 2006;44:336–43.CrossRefPubMed

38.

Ali H, Kabir N, Muhammad A, Shah MR, Musharraf SG, Iqbal N, et al. Hautriwaic acid as one of the hepatoprotective constituent of Dodonaea viscosa. Phytomedicine. 2014;21:131–40.CrossRefPubMed

39.

Liu C, Tao Q, Sun M, Wu JZ, Yang W, Jian P, et al. Kupffer cells are associated with apoptosis, inflammation and fibrotic effects in hepatic fibrosis in rats. Lab Invest. 2010;90:1805–16.CrossRefPubMed

40.

Li GS, Jiang WL, Tian JW, Qu GW, Zhu HB, Fu FH. In vitro and in vivo antifibrotic effects of rosmarinic acid on experimental liver fibrosis. Phytomedicine. 2010;17:282–8.CrossRefPubMed

Title: Hepatoprotective effects of Micromeria croatica ethanolic extract against CCl4–induced liver injury in mice
Authors: Sanda Vladimir-Knežević
Olga Cvijanović
Biljana Blažeković
Marija Kindl
Maja Bival Štefan
Robert Domitrović
Publication date: 01-12-2015
Publisher: BioMed Central
Published in: BMC Complementary Medicine and Therapies / Issue 1/2015
Electronic ISSN: 2662-7671
DOI: https://doi.org/10.1186/s12906-015-0763-8

At a glance: The STEP trials

Springer Medicine

Hepatoprotective effects of Micromeria croatica ethanolic extract against CCl₄–induced liver injury in mice

Abstract

Background

Methods

Results

Conclusion

At a glance: The STEP trials

Springer Medicine

Abstract

Background

Methods

Results

Conclusion

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