HYPOLIPIDEMIC AND ANTI-FATTY LIVER EFFECTS EXERTED BY STANDARDIZED PUNICA GRANATUM L. PEEL EXTRACT IN HEPG2 CELL-LINE AND HIGH-FAT DIET-INDUCED MICE

  • Omar A. Taha Biomedical research lab, Faculty of Pharmacy, Heliopolis University for Sustainable Development , 3 Cairo-Belbeis Road, El Horreya, Cairo, P.B. 2834, Egypt
  • Alaa Barakat
  • Heba K. Abdelhakim
  • Mohamed Abbas Shemis
  • Zainab Zakaria

Abstract

Objective: Pomegranate, (Punica granatum L., Lythraceae) peel has concentrated amounts of lipid-lowering elements that demonstrated, in various hoary and recent studies, their effects against obesity and hyperlipidemia, which involves elevated rates of lipid and lipoprotein levels in blood and increases risks of cardiovascular diseases.

We aim to study expression modulation of genes involved in lipid metabolism by the impact of standardized pomegranate peel extract (PPE) in a comprehensive research on human liver cells and experimental mice.

Methods: Using reverse-transcription real-time PCR, an in vitro study harnessing HepG2 cell line was conducted to determine the hyperlipidemia-related gene expression profiles and cytotoxic effects upon treatment with PPE. In another complementary in vivo study, male C57BL/6J mice were fed a high-fat diet (HFD) or an HFD supplemented with PPE for 14 d to define the expression of lipid metabolism related genes that control obesity. Fatty liver proportions were also estimated after treatment.

Results: Higher mRNA expression of LDL receptor (LDL-R) and down-regulation of sterol regulatory element-binding protein (SREBF-2), (SRBEP-1c), Fatty acid synthase (FAS) and 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) upon PPE treatment in HepG2 cell line were significantly recorded. In vivo study indicated significant weight reduction of body and liver, besides amelioration of fatty liver state detected by histological analysis. Moreover, the reverse-transcription real-time PCR assay demonstrated suppression (FAS) expression and up regulation of hormone sensitive lipase (HSL) in mice isolated liver and white adipose tissues.

Conclusion: Our study manages to affirm the hypolipidemic and anti-fatty liver influence of Punica granatum L. peel extract, reflected by molecular evaluation above and beyond other physiological assays.

Keywords: Pomegranate, Peel extract, Hyperlipidemia, LDLR, SREBP, FAS, HMGCR, HSL

Keywords: Punica granatum L. peel extract, hyperlipidemia, LDLR, SREBP, FAS, HMGCR, HSL

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References

Joo NS, Kim SM, Kim KM, Kim CW, Kim BT, Lee DJ. Changes of body weight and inflammatory markers after 12-week intervention trial: results of a double-blind, placebo-control pilot study. Yonsei Med J 2011;52:242-8.
2. Ono M, Okamoto N, Saibara T. Prospect of treatment based on pathogenesis and epidemiology. Adiposcience 2010;7:68-75.
3. Eliot LA, Jamali F. Pharmacokinetics, and pharmacodynamics of nifedipine in untreated and atorvastatin-treated hyper-lipidemic rats. J Pharmacol Exp Ther 1999;291:188-93.
4. Wasan KM, Brocks DR, Lee SD, Sachs-Barrable K, Thornton SJ. The impact of lipoproteins on the biological activity and disposition of hydrophobic drugs: implications for drug discovery. Nat Rev Drug Discovery. 2008;7:84-99.
5. Wasan KM, Looije NA. Emerging pharmacological approaches to the treatment of obesity. J Pharm Pharm Sci 2005;8:259-71.
6. Genest J, Frohlich J, Fodor G, McPherson R, Working Group on H, Other D. Recommendations for the management of dyslipidemia and the prevention of cardiovascular disease: summary of the 2003 update. Can Med Assoc J 2003;169:921-4.
7. Austin MA. Epidemiology of hypertriglyceridemia and cardiovascular disease. Am J Cardiol 1999;83:13F-6F.
8. Manolio TA, Pearson TA, Wenger NK, Barrett-Connor E, Payne GH, et al. Cholesterol and heart disease in older persons and women. Review of an NHLBI workshop. Ann Epidemiol 1992;2:161-76.
9. Ignat'eva EV, Merkulova TI, Vishnevskii OV, Kel AE. [Transcriptional regulation of lipid metabolism genes: description in the TRDD database]. Molekuliarnaia Biologiia 1997;31:684-700.
10. Rea TJ, DeMattos RB, Pape ME. Hepatic expression of genes regulating lipid metabolism in rabbits. J Lipid Res 1993; 34:1901-10.
11. de Jonge JD, Pennings JL, Baken KA, Konings J, Ezendam J, Van Loveren H. Gene expression changes in the mesenteric lymph nodes of rats after oral peanut extract exposure. J Immunotoxicol 2008;5:385-94.
12. Kim H, Bartley GE, Rimando AM, Yokoyama W. Hepatic gene expression related to lower plasma cholesterol in hamsters fed high-fat diets supplemented with blueberry peels and peel extract. J Agric Food Chem 2010;58:3984-91.
13. Zhang Y, Li Q, Wan HY, Helferich WG, Wong MS. Genistein and a soy extract differentially affect three-dimensional bone parameters and bone-specific gene expression in ovariectomized mice. J Nutr 2009;139:2230-6.
14. Middha SK, Usha T, Pande V. A review on antihyperglycemic and anti hepatoprotective activity of eco-friendly punica granatum peel waste. evidence-based complementary and alternative medicine. J Evidence-Based Complementary Altern Med 2013. Doi.org/10.1155/2013/656172. [Article in Press]
15. Priyanka B, Mohd A, Vidhu A, Malay B, Shahnaz S. Antidiabetic effect of punica granatum flowers: effect on hyperlipidemia, pancreatic cells lipid peroxidation and antioxidant enzymes in experimental diabetes. Food Chem Toxicol 2009;47:50-4.
16. Jafri MA, Aslam M, Javed K, Singh S. Effect of punica granatum Linn. (flowers) on blood glucose level in normal and alloxan-induced diabetic rats. J Ethnopharmacol 2000;70:309-14.
17. Kaplan M, Hayek T, Raz A, Coleman R, Dornfeld L, Vaya J, et al. Pomegranate juice supplementation to atherosclerotic mice reduces macrophage lipid peroxidation, cellular cholesterol accumulation and development of atherosclerosis. J Nutr 2001;131:2082-9.
18. Lei F, Zhang XN, Wang W, Xing DM, Xie WD, Su H, et al. Evidence of anti-obesity effects of the pomegranate leaf extract in high-fat diet induced obese mice. Int J Obes 2007;31:1023-9.
19. Aguilar CN, Aguilera-Carbo A, Robledo A. Production of antioxidant nutraceuticals by solid-state cultures of pomegranate (Punica granatum) peel and creosote bush (Larrea tridentata) leaves. Food Technol Biotechnol 2008;46:218–22.
20. Middha SK, Usha T, Pande V. A review on antihyperglycemic and anti hepatoprotective activity of eco-friendly punica granatum peel waste. J Evidence-Based Complementary Altern Med 2013. Doi.org/10.1155/2013/656172. [Article in Press]
21. Cheng S, Guo C, Yang J, Wei J, Li Y, Xu J. Experimental study on the hypolipidemic effect of polyphenolic extract from pomegranate peel. J Prev Med Chin People's Liberation Army 2004;3:160–3.
22. Anoosh E, Mojtaba E, Fatemeh S. Antioxidant activity of juice and peel extract of three variety of pomegranate and the effect of pomegranate juice on the plasma lipids. Int J Biosci 2012;2:116-23.
23. Abdel Motaal A, Shaker S. Anticancer and antioxidant activities of standardized whole fruit, pulp, and peel extracts of egyptian pomegranate. Open Conf Proc J 2011;211:41-5.
24. Shin J, Lee HJ, Jung DB, Jung JH, Lee HJ, Lee EO, et al. Suppression of STAT3 and HIF-1 alpha mediates antiangiogenic activity of betulinic acid in hypoxic PC-3 prostate cancer cells. Plos One 2011;6:e21492. Doi:10.1371/journal.pone.0021492. [Article in Press]
25. The Council European Economic Community Directive of 24 November 1986 on the approximation of laws, regulations and administrative provisions of the Member States regarding the protection of animals used for experimental and other scientific purposes (86/609/EEC); 1986.
26. Folch J, Lees M, Sloane-Stanley GH. A simple method for the isolation and purification of total lipides from animal tissues. J Biol Chem 1957;226:497–509.
27. Bancroft JD, Stevens A, Dawswon MP. Theory and practice of histological techniques. 4th ed. Churchill Livingstone, Edinbergh, London and New York; 1996. p. 273-92.
28. Girija K, Lakshman K. Antihyperlipidemic activity of methanol extracts of three plants of amaranthus in triton-WR 1339 induced hyperlipidemic rats. Asian Pac J Trop Dis 2011;1:1-4.
29. Pande V, Dubey S. Antihyperlipidemic activity of Sphaeranthus indicus on atherogenic diet induced hyperlipidemia in rats. Int J Green Pharm 2009;2:159-61.
30. Adisakwattana S, Chanathong B. Alpha-glucosidase inhibitory activity and lipid-lowering mechanisms of moringa oleifera leaf extract. Eur Rev Med Pharmacol Sci 2011;15:803-8.
31. Rahbar AR, Nabipour I. The hypolipidemic effect of Citrullus colocynthis on patients with hyperlipidemia. Pak J Biol Sci 2010;13:1202-7.
32. Bavarva JH, Narasimhacharya AVRL. Antihyperglycemic and hypolipidemic effects of costus speciosus in alloxan induced diabetic rats. Phytother Res 2008;22:620-6.
33. Vijayaraj PS, Muthukumar K, Sabarirajan J, Nachiappan V. Evaluation of antihyperlipidemic activity of ethanolic extract of Cassia auriculata flowers. Indian J Biochem Biophys. 2011;48:54-8.
34. Hajhashemi V, Abbasi N. Hypolipidemic activity of anethum graveolens in rats. Phytother Res 2008;22:372–5.
35. Brahma Srinivas Rao Desu, Saileela CH. The anti-hyperlipidemic activity of methanolic extract of rhinacanthus nasutus. Int J Res Pharm Chem 2013;3:708-11.
36. Nishant P. Ameliorative effects of herbal combinations in hyperlipidemia. Oxid Med Cell Longevity 2011:160-408.
37. Singh K, Chander R, Kapoor NK. Guggulsterone, a potent hypolipidaemic, prevents oxidation of low-density lipoprotein. Phytother Res 1997;11:291-4.
38. Liu JF, Ma Y, Wang Y, Du ZY, Shen JK, Peng HL. Reduction of lipid accumulation in HepG2 cells by luteolin is associated with activation of AMPK and mitigation of oxidative stress. Phytother Res 2011;25:588-96.
39. Kumar V, Fausto N, Abul Abbas. Robbins and cotran pathological basics of disease. 7 ed. Elsevier; 2004.
40. Kumar K, Vijayalakshmi K. In vitro antioxidant activity and cytotoxic potential of punica granatum peel and Vitis vinifera seeds on hep g2 cell lines. Int J Future Biotechnol 2013;1:1-7.
41. Sudhof TC, Goldstein JL, Brown MS, Russell DW. The LDL receptor gene: a mosaic of exons shared with different proteins. Science 1985;228:815-22.
42. Kovanen PT. Regulation of plasma cholesterol by hepatic low-density lipoprotein receptors. Am Heart J 1987;113:464-9.
43. Brown MS, Herz J, Goldstein JL. LDL-receptor structure. Calcium cages, acid baths, and recycling receptors. Nature 1997;388:629-30.
44. Yokoyama C, Wang XD, Briggs MR, Admon A, Wu J, Hua XX, et al. Srebp-1, a basic-helix-loop-helix-leucine zipper protein that controls transcription of the low-density lipoprotein receptor gene. Cell 1993;75:187-97.
45. Hua X, Yokoyama C, Wu J, Briggs MR, Brown MS, Goldstein JL, et al. SREBP-2, a second basic-helix-loop-helix-leucine zipper protein that stimulates transcription by binding to a sterol regulatory element. Proc Natl Acad Sci USA 1993;90:11603-7.
46. Gunstone FD, Harwood L, Dijkstra. The lipid handbook with Cd-Rom. Boca Raton: 3 ed. CRC Press; 2007.
47. Lewis JR, Mohanty SR. Nonalcoholic fatty liver disease: a review and update. Dig Dis Sci 2010;55:560-78.
48. Portincasa P, Grattagliano I, Palmieri VO, Palasciano G. Nonalcoholic steatohepatitis: recent advances from experimental models to clinical management. Clin Biochem 2005;38:203-17.
49. Xu KZ, Zhu C, Kim MS, Yamahara J, Li Y. Pomegranate flower ameliorates fatty liver in an animal model of type 2 diabetes and obesity. J Ethnopharmacol 2009;123:280-7.
50. Jang A, Srinivasan P, Lee NY, Song HP, Lee JW, Lee M, et al. Comparison of hypolipidemic activity of synthetic gallic acid linoleic acid ester with a mixture of gallic acid and linoleic acid, gallic acid, and linoleic acid on high-fat diet induced obesity in C57BL/6 Cr Slc mice. Chem Biol Interact 2008;174:109-17.
51. Zang Y, Zhang L, Igarashi K, Yu C. The anti-obesity and anti-diabetic effects of kaempferol glycosides from unripe soybean leaves in high-fat-diet mice. Food Function 2015;6:834-41.
52. Jadhav R, Puchchakayala G. Hypoglycemic and antidiabetic activity of flavonoids: boswellic acid, ellagic acid, quercetin, rutin on streptozotocin-nicotinamide induced type 2 diabetic rats. Int J Pharm Pharm Sci 2012;4:251-6.
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Taha, O. A., A. Barakat, H. K. Abdelhakim, M. A. Shemis, and Z. Zakaria. “HYPOLIPIDEMIC AND ANTI-FATTY LIVER EFFECTS EXERTED BY STANDARDIZED PUNICA GRANATUM L. PEEL EXTRACT IN HEPG2 CELL-LINE AND HIGH-FAT DIET-INDUCED MICE”. International Journal of Pharmacy and Pharmaceutical Sciences, Vol. 8, no. 6, Apr. 2016, pp. 156-61, https://innovareacademics.in/journals/index.php/ijpps/article/view/11226.
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