Annona muricata prevent hepatic fibrosis by enhancing lysosomal membrane stability and suppressing extracellular matrix protein accumulation


  • Usunomena Usunobun benson idahosa university, benin city, edo state, nigeria
  • Ngozi Okolie University of Benin





Annona muricata, Dimethylnitrosamine, Extracellular matrix, Fibrosis, Liver


Background: Hepatic fibrosis is a pre-pathologic state of cirrhosis, which plays a pivotal role in the pathogenesis of hepatocellular carcinoma (HCC). This study was carried out to evaluate the hepatoprotective effect of Annona muricata leaves on extracellular matrix (ECM) accumulation, lysosomal membrane integrity and liver damage in dimethylnitrosamine (DMN)-induced fibrotic rats.

Methods: Healthy adult male wistar rats were divided into four groups. Group I rats received normal saline and served as control, Group II rats were administered 200mg/kg Annona muricata, Group III received 200mg/kg Annona muricata orally simultaneously with 10mg/kg DMN, Group IV received 10 mg/kg DMN without extract. DMN administered intraperitoneally was on first three days of each week for two weeks while Annona muricata was administered for 14 consecutive days. 24 hours after last administration, all animals were sacrificed and blood collected for serum analysis. Biochemical parameters such as lactate dehydrogenase (LDH), total bilirubin (TB), total protein (TP), and albumin (ALB) were estimated to assess liver function and synthetic ability while hyaluronic acid (HA) was determined to assess extracellular matrix (ECM) function. In addition, acid phosphatase (ACP) was estimated to assess lysosomal membrane stability/fragility.

Results: Liver damage, decreased synthetic ability, lysosomal membrane fragility and altered ECM function were evident by an increase in the levels of LDH, TB, ACP and HA as well as a decrease in level of TP and ALB in sera of rats administered DMN compared to normal rats. However, simultaneous treatment with Annona muricata leaf extract significantly (p<0.05) reversed alterations in the above indices.

Conclusion: This study suggests that Annona muricata leaf functions as a potent fibrosuppressant by suppressing ECM accumulation, enhancing lysosomal membrane stability and liver synthetic ability.


[1] Ahmad R, Ahmed S, Khan NU & Hasnain A (2009). Operculina turpethum attenuates N-nitrosodimethylamine induced toxic liver injury and clastogenicity in rats. Chemical and Biological Interaction 181, 145–53.

[2] Ajmire PV, Chidambaranathan N, Dewade DR, Narkhede MB, & Wagh AE (2011). Effect of Boerhaavia diffusa against Dimethylnitrosamine induced liver cirrhosis. International Journal of Pharmacy and Pharmaceutical Sciences, 3(5), 366-370

[3] Akanji MA, Nafiu MO & Yakubu MT (2008). Enzyme Activities and Histopathology of Selected Tissues in Rats Treated with Potassium bromate. African Journal of Biomedical Research 11, 87 – 95.

[4] Arthur FKN, Woode E, Terlabi EO & Larbie C (2011). Evaluation of acute and subchronic toxicity of Annona Muricata (Linn.) Aqueous extract in animals. European Journal of Experimental Biology 1, 115-124.

[5] Chang FR (2001). Novel cytotoxic annonaceous acetogenins from Annona muricata, Journal National Productivity 64, 925–931.

[6] Collins AJ & Lewis DA (1971). Lysosomal enzyme level in blood of arthritic rats. Biochemistry and Pharmacology 28, 251-253.

[7] Di-Stasi LC; & Hiruma-Lima CA (2002). Plantas Medicinais na Amazônia e na Mata Atlântica, 2nd ed.; Editora UNESP: São Paulo, Brazil 87-112.

[8] Fried MW, Duncan A, & Soroka S (2001). Serum hyaluronic acid in patients with veno-occlusive disease following bone marrow transplantation. Bone Marrow Transplantation 27(6), 635-9.

[9] George J & Chandrakasan G (1997). Lactate dehydrogenase isoenzymes in dimethylnitrosamine induced hepatic fibrosis in rats. Journal of Clinical Biochemistry and Nutrition 22, 51–62.

[10] George J, Rao KR, Stern R, & Chandrakasan G (2001). Dimethylnitrosamine-induced liver injury in rats: the early deposition of collagen. Toxicology 156(2-3), 129-138.

[11] George J & Stern R (2004). Serum hyaluronan and hyaluronidase: very early markers of toxic liver injury. Clinica. Chimica. Acta. 348, 189–197.

[12] Greige-Gerges H, Khalil RA, Chahine R, Haddad C & Harb W (2007). Effect of cucurbitacins on bilirubin-albumin binding in human plasma. Life Sciences 80, 579-585.

[13] Halfon P, Bourliere M, & Penaranda G (2005). Accuracy of hyaluronic acid level for predicting liver fibrosis stages in patients with hepatitis C virus. Complementary Hepatology 4, 6-12.

[14] Hasegawa T, Sasaki T, Kimura T, Hoki M, Okada A, Mushiake S, Yagi M, & Imura K (2000). Measurement of serum hyaluronic acid as a sensitive marker of liver fibrosis in biliary atresia. Journal of Pediatric Surgery 35, 1643–1646.

[15] Huang, W, Zhang J, Chua SS, Qatanani M & Han Y (2003). Induction of bilirubin clearance by the Constitutive Androstane Receptor (CAR). PNAS 100, 4156-4161.

[16] Liaw CC, Chang FR, Lin CY, Chou CJ, Chiu HF, Wu MJ, & Wu YC (2002). New cytotoxic monotetrahydrofuran annonaceous acetogenins from Annona muricata. Journal of National Productivity 65, 470–475.

[17] Muthulingam M, Mohandoss P, Indra N, & Sethupathy S (2010). Antihepatotoxic efficacy of Indigofera tinctoria on paracetamol-induced liver damage in rats. International Journal of Pharmacy and Biomedical Research 1(1), 13-18.

[18] Nandan D, Venkatesan K, Katoch K, & Dayal RS (2007). Serum beta-glucuronidase levels in children with leprosy. Leprosy Rev. 78, 243-247.

[19] Nanji AA, Tahan S R, Khwaja S, Yacoub LK & Sadrzadeh SM (1996). Elevated plasma levels of hyaluronic acid indicate endothelial cell dysfunction in the initial stages of alcoholic liver disease in the rat. Journal of Hepatology 24, 368–374.

[20] Nyberg A, Engstrom-Laurent A & Loof L (1988). Serum hyaluronate in primary biliary cirrhosis--a biochemical marker for progressive liver damage. Hepatology 8(1), 142-6.

[21] Ostrow JD, Pascolo L, Shapiro SM & Tiribelli C (2003). New concepts in bilirubin encephalopathy. European Journal of ClinicalInvestment 33, 988-997.

[22] Patel K, Lajoie A, Heaton S, Pianko S, Behling CA, Bylund D, Pockros PJ, Blatt LM, Conrad A, & McHutchison JG (2003). Clinical use of hyaluronic acid as a predictor of fibrosis change in hepatitis C. Journal of Gastroenterology Hepatology 18, 253–257.

[23] Premila A (2004). Lysosomal enzyme activity during development of Carbon tetrachloride induced cirrhosis in rats. Indian Journal of Physiology Pharmacology 48 (2), 206–212

[24] Qing-Wei H. & Geng-Tao L. (2006). Effects of bicyclol on dimethylnitrosamine-induced liver fibrosis in mice and its mechanism of action. Life Sciences 79, 606–612.

[25] Rajadurai M & Prince PS (2007). Preventive effect of naringin on cardiac mitochondrial enzymes during isoproterenol-induced myocardial infarction in rats: a transmission electron microscopic study. Journal of Biochemistry and Molecular Toxicology 21(6), 354-61.

[26] Sandya S, Achan MA & Sudhakaran PR (2007). Parallel changes in fibronectin and α5β1 integrin in articular cartilage in type II collagen- induced arthritis. Indian Journal of Biochemistry and Biophysics 44(1), 14-18

[27] Shin M & Moon J (2010). Effect of dietary supplementation of grape skin and seeds on liver fibrosis induced by dimethylnitrosamine in rats. Nutrition Research and Practice 4(5), 369-374.

[28] Skripenova S, Trainer TD, & Krawitt EL (2007). Variability of grade and stage in simultaneous paired liver biopsies in patients with hepatitis C. Journal of Clinical Patholology 60(3), 321-4.

[29] Sousa, MP, Matos MEO, Matos FJA, Machados MIL, & Craveiro AA (2004). Constituintes Químicos Ativos e Propriedades Biológicas de Plantas Medicinais Brasileiras, 2nd ed.; Editora UFC: Fortaleza, Brazil 281-283.

[30] Stickel F, Urbaschek R, Schuppan D, Poeschl G, Oesterling C, Conradt C, McCuskey RS Simanowski UA & Seitz HK (2001). Serum collagen type VI and XIV and hyaluronic acid as early indicators for altered connective tissue turnover in alcoholic liver disease. Digest. Disease Sciences 46 (9), 2025–2032.

[31] Taylor L (2002). Graviola (Annona muricata). In Herbal Secrets of the Rainforest 2nd ed. Roseville, CA: Prima Publishing.

[32] Usunobun U & Okolie NP (2015). Phytochemical analysis and mineral composition of Annona muricata leaves. International Journal of Research and Current Development 1(1), 38-42.

[33] Usunobun U, Okolie NP, Anyanwu OG, Adegbegi AJ & Egharevba ME (2015). Phytochemical screening and proximate composition of Annona muricata leaves. European Journal of Botany, Plaint Science and Phytology 2(1), 18-28.

[34] Usunobun U, Okolie PN & Eze GI (2015a). Modulatory effect of ethanolic leaf extract of Annona muricata pre-treatment on liver damage induced by Dimethylnitrosamine (DMN) in rats. British Journal of Pharmaceutical Research 8(3), 1-9.

[35] Usunobun U, Okolie NP, & Eze IG. (2015b). Attenuation of N, N-Dimethylnitrosamine-induced liver Fibrosis in rats by ethanolic leaf extract of Annona muricata. Saudi Journal of Medical and Pharmaceutical Sciences 1(2), 62-69.

[36] Vogel AI (1971). A textbook of practical organic Chemistry including qualitative organic analysis. Longman group limited, London. Pp 426.

[37] Vrochides D, Papanikolaou V, Pertoft H, Antoniades AA, & Heldin, P (1996). Biosynthesis and degradation of hyaluronan by nonparenchymal liver cells during liver regeneration. Hepatology 23, 1650–1655.

[38] Wyatt HA, Dhawan A, & Cheeseman P (2002). Serum hyaluronic acid concentrations are increased in cystic fibrosis patients with liver disease. Archive Disease and Childhealth 86(3), 190-3.

[39] Yacout GA, Elguindy NM & El-Azab EF (2012). Hepatoprotective effect of basil (Ocimum basilicum L.) on CCl4-induced liver fibrosis in rats. Afriican Journal of Biotechnology 11(90), 15702-15711.

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