• Nagwa E. Awad Pharmacognosy Department National Research Centre, 33 El-Bohouth St., Dokki, Cairo, Egypt. ID: 60014618
  • Mostafa A. Abdelkawy Pharmacognosy Department, Faculty of Pharmacy, Cairo University, Cairo, Egypt
  • Manal A. Hamed Therapeutic Chemistry Department National Research Centre, 33 El-Bohouth St., Dokki, Cairo, Egypt. ID: 60014618
  • Ahmed M. A. Souleman Phytochemistry and Plant Systematic Department, National Research Centre, 33 El-Bohouth St., Dokki, Cairo, Egypt. ID: 60014618
  • Enas H. Abdelrahman Pharmacognosy Department, Faculty of Pharmacy, Cairo University, Cairo, Egypt
  • Nehal S. Ramadan Pharmacognosy Department National Research Centre, 33 El-Bohouth St., Dokki, Cairo, Egypt. ID: 60014618


Objective: The antioxidant and hepatoprotective activities of ethyl acetate (EA) fraction of the dried aerial part of Justicia spicigera were evaluated and the characterization of its anthocyanin content was done.

Methods: Hepatic fibrosis was induced by carbon tetrachloride (CCl4) in rats. The ethyl acetate fraction was obtained by successive liquid/liquid fractionation of the crude cold ethanolic extract and the pigments were characterized by HPLC technique. The in vitro studies were carried out through evaluation of the EA fraction on the attenuation of 1, 1-Diphenyl-2-picrylhydrazyl (DPPH) free radicals. The in vivo biological evaluation was done in CCl4 injured rats through determination of liver function indices, oxidative stress markers and the histopathological picture of the treated liver.

Results: The phenolic content in the EA fraction was 42.94 mg/g. Twelve anthocyanins were identified, the major of which are peonidin 3, 5-diglucoside (64.30%), malvidin 3, 5-diglucoside (10.59%) and petunidin 3,5-diglucoside (4.71%). Treatment of CCl4 intoxicated rats with EA fraction recorded improvement in the liver function indices and oxidative stress markers. The histopathological observations confirmed our results.

Conclusion: The ethyl acetate fraction of the dried aerial part of Justicia spicigera recorded antioxidant and hepato protective activities.


Keywords: Justicia spicigera, Acanthaceae, Phenolic content, Anthocyanins, Oxidative stress, Histopathology


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1. Guven A, Guven U, Gulmez M. The effect of kefir on the activities of GSH, GSHPX, GST, CAT, GSH and LPO levels in carbon tetrachloride-induced mice tissue. J Vet 2003;50:412-6.
2. Ozturk F, Ucar M, Ozturk IC, Vardi N, Batcioglu K. Carbon tetrachloride induced nephrotoxicity and protective effect of betaine in Sprague Dawley rats. Urology 2003;62:353-6.
3. Khan RA, Khan MR, Sahreen S, Shah NA. Hepatoprotective activity of Sonchus asper against carbon tetrachloride-induced injuries in male rats: a randomized controlled trial. BMC Complementary Altern Med 2012;12:90.
4. Motawi TK, Hamed MA, Shabana MH, Hashem RM, Aboul Naser AF. Zingiber officinale acts as a nutraceutical agent against liver fibrosis. Nutr Metab 2011;8:40-50.
5. Khan MR, Rizvi W, Khan GN, Khan RA, Shaheen S. Carbon tetrachloride induced nephrotoxicity in rat: Protective role of Digera muricata. J Ethnopharmacol 2009;122:91-9.
6. Weber LW, Boll M, Stampfl A. Hepatotoxicity and mechanism of action of haloalkanes: carbon tetrachloride as a toxicological model. Carr Rev Toxicol 2003;33:105-36.
7. Ichi I, Kamikawa C, Nakagawa T, Kobayashi K, Kataoka R, Nagata E, et al. Neutral sphingomyelinase-induced ceramide accumulation by oxidative stress during carbon tetrachloride intoxication. Toxicology 2009;261:33-40.
8. Jia X, Han C, Chen J. Effect of tea on preneoplastic lesions and cell cycle regulators in rat liver. Cancer Epidemiol Biomarkers Prev 2002;11:1663–7.
9. Van Gijssel HE, Maassen CB, Mulder GJ, Meerman JH. P53 protein expression by hepatocarcinogens in the rat liver and its potential role in mitoinhibition of normal hepatocytes as a mechanism of hepatic tumor promotion. Carcinogenesis 1997;18:1027–32.
10. El-Gengaihi S, Hamed M, Hassan E, Zahran H, Arafa M. Chemical composition and nutritional effect of Physalis peruviana husk as hepato-renal protective agent. Int J Phytomed 2012;4:229-36.
11. El-Gengaihi SE, Hassan EE, Hamed MA, Zahran HG, Mohammed MA. Chemical composition and biological evaluation of Physalis peruviana root as hepato-renal protective agent. J Diet Suppl 2013;10:39–53.
12. Opoku AR, Ndlovu IM, Terblanche SE, Hutchings AH. In vivo hepatoprotective effects of Rhoicissus tridentata subsp. cuneifolia, a traditional Zulu medicinal plant, against CCl4-induced acute liver injury in rats. South Afr J Botany 2007;73:372-7.
13. Braz DM, Carvalho-Okano RM, Kameyama C. Acanthaceae da reserva florestal matadoparaı´so. Vic-osa, Minas Gerais. Rev Bras Bot 2002;25:495–504.
14. Arellano-Rodrı´guez JA, Flores Guido JS, Tun J, Garridoy MM, Cruz B. Nomenclatura, formadevida, uso, manejoy distribucion de la sespecies vegetales de la penınsula de Yucatan. Etnoflora Yucatanense 2003;20:5-6.
15. Ortiz-Andrade R, Cabanas-Wuan A, Arana-Argaez VE, Alonso-Castro AJ, Zapata-Bustos R, Salazar-Olivo LA, et al. Anti diabetic effects of Justicia spicigera Schltdl (Acanthaceae). J Ethnopharmacol 2012;143:455–62.
16. Singleton VL, Rossi JA. Colorimetry of total phenolics with phosphomolybdic phosphotungstic acid reagents. Am J Enol Vitic 1965;16:144-58.
17. Watada AE, Abbott JA. Objective method of estimating anthocyanin content for determining color grade of grapes. J Food Sci 1975;40:1276-9.
18. Chen HY, Lin YC, Hsiech CL. Evaluation of antioxidants activity of aqueous extract of some selected nutraceutical herbs. Food Chem 2007;104:1418–24.
19. Marsillach J, Camps J, Ferré N, Beltran R, Rul A, Mackness B, et al. Paraoxonase-1 is related to inflammation, fibrosis and PPAR delta in experimental liver disease. BMC Gastroenterol 2009;9:3.
20. Ibarra-Moreno S, Ibarra-Velarde F, Avila-Acevedo JG. Obtaining the minimum lethal dose against Fasciola hepatica in vitro using plant extract hexanes with fasciolicide activity and toxicity evaluation on CD1 male mice. Am J Plant Sci 2012;3:899-903.
21. Yuvaraj P, Subramoniam A. Hepatoprotective property of Thespesia populnea against carbon tetrachloride induced liver damage in rats. J Basic Clin Physiol Pharmacol 2009;20:169-77.
22. Buege JA, Aust SD. Microsomal lipid peroxidation. Methods Enzymol 1978;52:302-10.
23. Moron MS, Depierre JW, Mannervik B. Level of glutathione, glutathione reductase and glutathone-S-transferase activities in rat lung and liver. Biochim Biophys Acta 1979;582:67-78.
24. Nishikimi M, Rae NA, Yagi K. The occurrence of superoxide anion in the action of reduced phenazine methosulphate and molecular oxygen. Biochem Biophys Res Commun 1972;46:849-53.
25. Reitman S, Frankel S. Colorimetric method for the determination of serum glutamic oxaloacetic and glutamic pyruvic transaminases. Am J Clin Pathol 1957;28:56-63.
26. Belfield A, Goldberg DM. Hydrolysis of adenosine-monophosphate by acid phosphatase as measured by a continues spectrophotometric assay. Enzyme 1971;12:561-6.
27. Szasz G. A kinetic photometric method for serum gamma glutamyl transpeptidase. Clin Chem 1969;15:124-36.
28. 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.
29. Hirsch C, Zouain CS, Alves JB, Goes AM. Induction of protective immunity and modulation of granulaomatous hypersensitivity in mice using PIII, an anionic fraction of Schistosoma mansoni adult worm. Parasitology 1979;115:21-8.
30. Toklu HZ, Tunali AT, Velioglu-Ogunc A, Ercan F, Gedik N, Keyer-Uysal M, et al. Silymarin, the antioxidant component of Silybum marianum, prevents sepsis-induced acute lung and brain injury. J Surg Res 2008;145:214-22.
31. Jakobek L, Seruga M, Medvidovic-Kosanodvic M, Novak I. Anthocyacnoinn content and antioxidant activity of various red fruit juices. Dtsch Lebensm-Rundsch 2007;103:58-64.
32. Pasko P, Barton H, Zagrodzki P, Gorinstein S, Fołta M, Zachwieja Z. Anthocyanins, total polyphenols and antioxidant activity in amaranth and quinoa seeds and sprouts during their growth. Food Chem 2009;115:994-8.
33. Miguel MG. Anthocyanins Antioxidant and/or anti-inflammatory activities. J Appl Pharm Sci 2011;01:7-15.
34. Recknagel RO, Glende EA, Dolak JA, Waller RZ. Mechanisms of carbon tetrachloride toxicity. Pharmacol Ther 1989;43:139-54.
35. Lin HM, Tseng HC, Wang CJ, Lin JJ, Lo CW, Chou FP. Hepatoprotective effects of Solanum nigrum Linn. Extract against CCl4-induced oxidative damage in rats. Chem Biol Int 2008;171:283-93.
36. Romero FJ, Bosch-Morell F, Romero MJ, Jareno EJ, Romero B, Marín N, et al. Lipid peroxidation products and antioxidants in human disease. Environ Health Perspect 1998;106:1229-34.
37. Yu J, Wang Y, Qian H, Zhao Y, Liu B, Fu C. Polyprenolsfrom Taxus chinensis var. mairei prevent the development of CCl4-induced liver fibrosis in rats. J Ethnopharmacol 2012;142:151-60.
38. Gowri Shankar GNL, Manavalan R, Venkappayya D, Raj CD. Hepatoprotective and antioxidant effects of Commiphora berryi (Arn) Engl bark extract against CCl4-induced oxidative damage in rats. Food Chem Toxicol 2008;46:3182-5.
39. Mansour HH, Hafez HF, Fahmy NM. Silymarin modulates cisplatin-induced oxidative stress and hepatotoxicity in rats. J Biochem Mol Biol 2006;39:656-61.
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How to Cite
Awad, N. E., M. A. Abdelkawy, M. A. Hamed, A. M. A. Souleman, E. H. Abdelrahman, and N. S. Ramadan. “ANTIOXIDANT AND HEPATOPROTECTIVE EFFECTS OF JUSTICIA SPICIGERA ETHYL ACETATE FRACTION AND CHARACTERIZATION OF ITS ANTHOCYANIN CONTENT”. International Journal of Pharmacy and Pharmaceutical Sciences, Vol. 8, no. 8, Aug. 2015, pp. 91-96,
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