HPLC Analysis and Protective Effect of Fractions from Ethanolic Extract of Calotropis procera (Ait.) R. Br Root Bark Against Diethylnitrosamine Induced-Hepatic Damage in Wistar Rats http://www.doi.org/10.26538/tjnpr/v7i7.26
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Abstract
Calotropis procera (Ait.) R.Br (Apocynaceae) is a medicinal plant known for its multiple therapeutic virtues including the management of liver diseases in Burkina Faso. This study aimed to evaluate the hepato-protective properties of the fractions of the ethanol extract of Calotropis procera roots barks while identifying chemical compounds in these extracts. Sterols, triterpene and phenolic compounds were sought and quantified in the fractions of the ethanol extract of Calotropis procera root bark by HPLC-DAD. The anti-hepatotoxic activity of fractions was evaluated using diethylnitrosamine (DEN) to induce acute liver inflammation in male Wistar rats. Serum biomarkers, alanine transaminase (ALAT), aspartate transaminase (ASAT), alkaline phosphatase (ALP), total proteins and Albumin were evaluated and the activities of antioxidant enzymes (Superoxide dismutase and catalase) as well as the level of malondialdehyde (MDA) were determined in the liver homogenate. The histopathological study of sections of livers pretreated with the fractions was also carried out. The evaluation of the anti-hepatotoxic activity showed that the n-butanol fraction and residual aqueous fraction had best effects on serum biomarkers (ALAT, ASAT, ALP) as well as on the activities of the antioxidant enzymes Superoxide dismutase and catalase and on hepatic tissue. Gallic acid, an antioxidant, antihepatotoxic, antifibrotic compound was identified in significant quantities in these two fractions at the respective contents of 17.92 ± 0.26 μg/mg and 18.76 ± 0.27 μg/mg of dry extract, suggesting
that this compound would participates in the hepatoprotective properties of these extracts.
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Ott JJ, Stevens GA, Groeger J, Wiersma ST. Global epidemiology of hepatitis B virus infection: new estimates of age-specific HBsAg seroprevalence and endemicity. Vaccine. 2012; 30(12):2212-2219. doi:10.1016/j.vaccine.2011.12.116
WHO. Prevention, care and treatment of viral hepatitis in the african region: framework for action, 2016 – 2020. WHO Regional Office for Africa, 2017. http://apps.who.int/iris.
Meda N, Tuaillon E, Kania D, Tiendrebeogo A, Pisoni A, Zida S, Bollore K, Medah I, Laureillard D, Moles JP, Nagot N, Nebie KY, Van de Perre P, Dujols P. Hepatitis B and C virus seroprevalence, Burkina Faso: a cross-sectional study. Bull. World Health Organ. 2018; 96(11):750-759. doi:
2471/BLT.18.208603
Qureshi H, Ali SS, Iqbal M, Siddiqui AA, Khan NA, Hamid SS. Aflatoxins and Hepatitis B, C Viral Associated Hepatocarcinogenesis. J Cell Sci Ther. 2014; 5 (5) : 179-185; DOI:10.4172/2157-7013.1000179
AlHomaid FA, Hassan OMM. Histopathological and biochemical studies in male Wistar albino rats injected with diethylnitrosamine and treated with Camel’s milk and Curcuma longa. Egypt. J. Chem. Environ. Health. 2016; 2 (2):548-565
Schwierz C, Thiry N, Van de Sande S, Gamil M, Nevens F, Colle I, Horsmans Y, Hulstaert F. Evaluation économique du traitement antiviral de l’hépatite B chronique en Belgique – Tome 2. Health Technology Assessment (HTA). Bruxelles: Centre federal d’expertise des soins de santé (KCE). 2011. Reports 157B. D/2011/10.273/28
Chute DF, Chung RT, Sise ME. Direct-acting antiviral therapy for hepatitis C virus infection in the kidney transplant recipient. Kidney Int. 2018; 93(3): 560–567; https://doi.org/10.1016/j.kint.2017.10.024
Mukazayire MJ, Minani V, Ruffo CK, Bizuru E, Stévigny C, Duez P. Traditional phytotherapy remedies used in SouthernRwanda for the treatment of liver diseases. J. Ethnopharmacol. 2011; 138 (2): 415–431; https://doi.org/10.1016/j.jep.2011.09.025
Ladoh YCF, Dibong SD, Nyegue MA, Djembissi TRP, Lenta NB, Mpondo ME, Yinyang J, Wansi JD. Activité antioxydante des extraits méthanoliques de Phragmanthera capitata (Loranthaceae) récoltée sur Citrus sinensis. J. Appl. Biosci. 2014; 84 (2014):7636–7643; http://dx.doi.org/10.4314/jab.v84i1.9
WHO. Who global report on traditional and complementary medicine 2019. WHO, 2019; https://apps.who.int/iris/handle/10665/312342
Tibiri A, Boria S, Traoré TK, Ouédraogo N, Nikièma A, Ganaba S, Compaoré JM, Ouédraogo I, Guissou IP, Carraz M. Countrywide Survey of Plants Used for Liver Disease Management by Traditional Healers in Burkina Faso. Front. Pharmacol. 2020; 11:563751. doi:
3389/fphar.2020.563751
Sombié EN, Hilou A, Ouédraogo N, N’do JYP, Paré D, Traoré TK, Ouédraogo GG, Tibiri A. Hepatoprotective Effects of Calotropis procera (Ait.) R. Br Root Bark Extracts against Diethylnitrosamine Induced Liver Injury in Rats. European J Med Plants. 2021; 32(11): 12-22, 2021 ; DOI:10.9734/EJMP/2021/v32i1130426
Misra HP, Fridovich I. The Role of Superoxide Anion in the Autoxidation of Epinephrine and a Simple Assay for Superoxide Dismutase. JBC. 1972; 247(10):3170-3175. https://doi.org/10.1016/S0021-9258(19)45228-9
Beers RF, Sizer IW. A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. JBC. 1952; 195 (1): 133-140. https://doi.org/10.1016/S0021- 9258(19)50881-X
Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxidation in animal tissues by thiobarbituric acid reaction. Anal. Biochem. 1979; 95 (2): 351-358. https://doi.org/10.1016/0003-2697(79)90738-3
Meharie BG, Amare GG, Belayneh YM. Evaluation of Hepatoprotective Activity of the Crude Extract and Solvent Fractions of Clutia abyssinica (Euphorbiaceae) Leaf Against CCl4-Induced Hepatotoxicity in Mice. J. Exp. Pharmacol. 2020; 2020:12 137–150
Ciulei I. Methodology for Analysis of Vegetable Drugs. Practical Manual on the Industrial Utilisation of Medicinal and Aromatic Plants. Bucharest, Romania, 1982; 1-62.
Boligon AA, Piana M, Kubiça TF, Mario DN, Dalmolin TV, Bonez PC, Weiblen R, Lovato L, Alves SH, Campos MMA, Athayde ML. HPLC analysis and antimicrobial, antimycobacterial and antiviral activities of Tabernaemontana catharinensis A. DC. J. Appl. Biomed. 2015; 13 (2015): 7–18.
http://dx.doi.org/10.1016/j.jab.2014.01.004
Boligon AA, Kubiça TF, Mario DN, Faccim de Brum T, Piana M, Weiblen R, Lovato L, Alves SH, Santos RCV, Alves CFS, Athayde ML. Antimicrobial and antiviral activity-guided fractionation from Scutia buxifolia Reissek extracts. Acta Physiol Plant (2013) 35:2229–2239. DOI
1007/s11738-013-1259-0
International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use. ICH Harmonised Tripartite Guideline: Validation of Analytical procedures: text and methodology. 2005; p. 1-13.
N’do JY, Hilou A, Ouédraogo N, Sombié EN, Traoré KT. Phytochemistry, Antioxidant, and Hepatoprotective Potential of Acanthospermum hispidum DC Extracts against Diethylnitrosamine-Induced Hepatotoxicity in Rats. Medecines, 2018; 42(5):1–13. doi: 10.3390/medicines5020042
Pradeep K, Mohan CVR, Gobianand K, Karthikeyan S. Protective effect of Cassia fistula Linn. On diethylnitrosamine induced hepatocellular damage and oxidative stress in ethanol pretreated rats. Biol Res. 2010; 43(1): 113-125. http://dx.doi.org/10.4067/S0716- 97602010000100013
Bilgic S, Dogan Z, Azirak S, Erdemli ME, Onderci M, Turk A, Ozer MK. Hepatoprotective effect of royal jelly, grape seed extract, and Lycium barbarum against diethylnitrosamine-induced liver toxicity in rats. Ann. Med. Res. 2018; 25(3): 342-348, DOI: 10.5455/annalsmedres.2018.03.040
Sombié EN, Ouédraogo N, Hilou A. Anti-inflammatory activity and phytochemistry of extracts from the root bark of Calotropis procera (ait.) R. Br. Hepatoprotective medicinal plant from Burkina Faso. Asian J Pharm Clin Res. 2020; 13 (10): 185-190. DOI: http://dx.doi.org/10.22159/ajpcr.2020.v13i10.39275
Zhu L, Li B, Liu X, Meng X. Hepatoprotective Effects of Triterpenoid Isolated from Schizandra chinensis against Acute Alcohol-Induced Liver Injury in Mice. Food Sci. Technol. Res. 2013; 19 (6): 1003–1009
Wang Y, Wang G, Yi X, Zhang J, He X. Hepatoprotective and Antioxidant Effects of Total Triterpenoids from Poria cocos. European J Med Plants. 2017; 21(2): 1-9, DOI: 10.9734/EJMP/2017/37908
Chavda R, Vadalia KR, Gokani R. Hepatoprotectrive and antioxidant activity of root bark of Calotropis procera R.Br (Asclepediaceae). Int. J. Pharmacol. 2010; 6 (6): 937-943; DOI:10.3923/ijp.2010.937.943
Marzouk AM. Hepatoprotective Triterpenes from Hairy Root Cultures of Ocimum basilicum L. Z Naturforsch C J Biosci. 2009; 64(3-4):201-209. DOI: 10.1515/znc-2009-3-409
Prasanth DSNBK, Rao AS, Prasad YR. Hepatoprotective Activity of Aralia racemosa L. and its Triterpenoid and Steroid Compounds against Paracetamol – Induced Liver Injury in Albino Wistar Rat. Glob. J. Med. Res. 2017; 17 (2).
Gutiérrez-Rebolledo GA, Siordia-Reyes AG, MeckesFischer M, Jiménez-Arellanes A. Hepatoprotective properties of oleanolic and ursolic acids in antitubercular drug-induced liver damage. Asian Pac J Trop Med. 2016; 9(7):644-51. doi: 10.1016/j.apjtm.2016.05.015
Wong H-S, Chen J-H, Leong P-K, Leung H-Y, Chan W-M, Ko KM. β-Sitosterol Protects against Carbon Tetrachloride Hepatotoxicity but not Gentamicin Nephrotoxicity in Rats via the Induction of Mitochondrial Glutathione Redox Cycling. mol. 2014; 19 (11):17649-17662. doi:10.3390/molecules191117649
Kim K-S, Yang HJ, Lee J-Y, Na Y-C, Kwon S-Y, Kim Y-C, Lee J-H, Jang H-J. Effects of β-sitosterol derived from Artemisia capillaris on the activated human hepatic stellate cells and dimethylnitrosamine-induced mouse liver fibrosis. BMC Complement Altern. Med. 2014; 14:363.
doi:10.1186/1472-6882-14-363
Feng S, Gan L, Yang CS, Anna B, Liu AB, Lu W, Shao P, Dai Z, Sun P, Luo Z. Effects of Stigmasterol and β-Sitosterol on Nonalcoholic Fatty Liver Disease in a Mouse Model: A Lipidomic Analysis. J. Agric. Food Chem. 2018; 66 (13):3417–3425. https://doi.org/10.1021/acs.jafc.7b06146
Ibrahim SRM, Mohamed GA, Shaala LA, Banuls LMY, Goietsenoven GV, Kiss R, Youssef DTA. New ursane-type triterpenes from the root bark of Calotropis procera. Phytochem. Lett. 2012; 5 (2012) 490-495. http://dx.doi.org/10.1016/j.phytol.2012.04.012
Kim Y-S, Li X-F, Kang K-H, Ryu B-M, Kim S-K. Stigmasterol isolated from marine microalgae Navicula incerta induces apoptosis in human hepatoma HepG2 cells. BMB Rep. 2014; 47(8):433-8. doi: 10.5483/bmbrep.2014.47.8.153
El-Ghffar EAA, El-Nashar HAS, Eldahshan OA, Singab ANB. GC-MS analysis and hepatoprotective activity of then hexane extract of Acrocarpus fraxinifolius leaves against paracetamol-induced hepatotoxicity in male albino rats. Pharm Biol. 2017; 55(1): 441–449. doi:
1080/13880209.2016.1246575
Afifi NA, Ibrahim MA, Galal MK. Hepatoprotective influence of quercetin and ellagic acid on thioacetamideinduced hepatotoxicity in rats. Can. J. Physiol. Pharmacol. 2018; 96 (6).
Jabbar AAJ, Alamri ZZ, Abdulla MA, AlRashdi AS, Najmaldin SK, Zainel MA. Sinapic Acid Attenuate Liver Injury by Modulating Antioxidant Activity and Inflammatory Cytokines in Thioacetamide-Induced Liver Cirrhosis in Rats. Biomedicines. 2023, 11, 1447. https://doi.org/10.3390/ biomedicines11051447
Nile SH, Ko EY, Kim DH, Keum Y-S. Screening of ferulic acid related compounds as inhibitors of xanthine oxidase and cyclooxygenase-2 with anti-inflammatory activity. Rev. Bras. Farmacogn. 2016 ; 26 (1) : 50–55; http://dx.doi.org/10.1016/j.bjp.2015.08.013
Latief U, Husain H, Mukherjee D, Ahmad R. Hepatoprotective efficacy of gallic acid during Nitrosodiethylamine-induced liver inflammation in Wistar rats. j. basic appl. zool. 2016; 76 : 31–41 ; http://dx.doi.org/10.1016/j.jobaz.2016.07.002
Rasool MK, Sabina EP, Ramya SR, Preety P, Patel S, Mandal N, Mishra PP, Samuel J. Hepatoprotective and antioxidant effects of gallic acid in paracetamol-induced liver damage in mice. J. Pharm. Pharmacol. 2010; 62 (5): 638–643, DOI 10.1211/jpp/62.05.0012
Koul A, Binepal G, Gangar SC. Impediment of diethylnitrosamine induced hepatotoxicity in male Balb/c mice by pretreatment with aqueous Azadirachta indica leaf extract. Indian J. Exp. Biol. 2007; 45(4):359-366.
Boro H, Usha T, Babu D, Chandana P, Goyal AK, Ekambaram H, Yusufoglu HS, Das S, Middha SK. Hepatoprotective activity of the ethanolic extract of Morus indica roots from Indian Bodo tribes. SN Appl. Sci. 2022; 4:49 ; https://doi.org/10.1007/ s42452- 021-04859-z.
Adebayo AH, Akpata OB, Yakubu OF. Hepatoprotective Effect of the Methanolic Leaf Extract of Annona senegalensis Pers. against 7, 12-dimethylbenz[a]anthracene (DMBA)-Induced Toxicity in Wistar rats. Trop J Nat Prod Res. 2022; 6(3):388-394. doi.org/10.26538/tjnpr/v6i3.15.