INSIGHTS INTO THE ROLE OF MORUS ALBA IN REVERSING OBESITY-ASSOCIATED HEPATIC STEATOSIS AND RELATED METABOLIC DISORDER IN RATS

  • Hanaa H. Ahmed
  • Fatehya M Metwally
  • Hend Rashad
  • Asmaa M Zaazaa

Abstract

ABSTRACT
Objective: The goal of the present study was to examine the viability of Morus alba (M. alba) ethanolic extract in repression of obesity-associated
hepatic steatosis and related metabolic disorder; dyslipidemia, hyperinsulinemia, and glycemic status.
Methods: Adult female albino rats were randomly assigned into four groups, eight rats each as follows: Group (1) control group received standard
rodent diet for 24 weeks. The other three groups administered high cholesterol diet for 12 weeks and served as obese group, M. alba-treated group,
and simvastatin-treated group.
Results: The current results showed an increment in thoracic circumference (TCX) and abdominal circumferences (AC) as well as body mass index
(BMI) in obese group. In addition, dyslipidemia, hyperinsulinemia, hyperglycemia, and insulin resistance have been elucidated in obese group.
Moreover, hepatic malondialdehyde (MDA), nitric oxide (NO), serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), and bilirubin
values were significantly increased in obese groups versus control group. On the other hand, administration of ethanolic extract of Morus alba or
simvastatin could significantly lessen BMI and in addition to improve dyslipidemia in obese group. Glucose, insulin levels, and insulin resistance value
in serum samples demonstrated a significant reduction in obese group upon treatment with M. alba ethanolic extract or simvastatin. Furthermore,
noticeable depletion in hepatic MDA, NO contents, serum ALT, AST activities, and serum bilirubin level was recorded as a result of treatment with
either ethanolic extract of M. alba or simvastatin. Histopathological examination of liver tissue showed ballooning degeneration in the hepatocytes
(hepatic steatosis) associated with inflammatory cells penetration in portal zone in obese group. Meanwhile, the treatment of obese groups with
ethanolic extract of M. alba or simvastatin was found to restore the structural organization of the liver.
Conclusion: The present findings provide a novel aspect for understanding of the role of M. alba against obesity-associated liver diseases and related
metabolic disorder. The mechanisms underlying these effects seem to depend on the hypolipidemic potential, anti-inflammatory property, and
antioxidant activity of its phytochemicals.
Keywords: Obesity, Morus alba, Dyslipidemia, Hyperinsulinemia, Hyperglycemia, Hepatic steatosis.

References

REFERENCES
1. Himaja N, Shama NS. Herbal wealth for hepatotoxicity: A review.
Asian J Pharm Clin Res 2015;8(1):3-9.
2. Wolf PL. Biochemical diagnosis of liver disease. Indian J Clin Biochem
1999;14(1):59-90.
3. Piyachaturawat P, Chai-ngam N, Chuncharunee A, Komaratat P,
Suksamrarn A. Choleretic activity of phloracetophenone in rats:
Structure-function studies using acetophenone analogues. Eur J
Pharmacol 2000;387(2):221-7.
4. Ward FM, Daly MJ. Hepatic disease. In: Clinical Pharmacy and
Therapeutics. Churchill Livingstone; 1999. p. 195-212.
5. Ahmed HH, Hamza AH, Kotob SE. Corn silk offers multimechanistic
approaches in mitigating obesity in rodents. Asian J Pharm Clin Res
2016;9(4):1-7.
6. Trayhurn P, Beattie JH. Physiological role of adipose tissue: White
adipose tissue as an endocrine and secretory organ. Proc Nutr Soc
2001;60(3):329-39.
7. Xu H, Barnes GT, Yang Q, Tan G, Yang D, Chou CJ, et al. Chronic
inflammation in fat plays a crucial role in the development of obesityrelated
insulin resistance. J
Clin
Invest 2003;112(2):1821-30.
8. Trayhurn P, Wood IS. Adipokines: Inflammation and the pleiotropic
role of white adipose tissue. Br J Nutr 2004;92(3):347-55.
9. Diehl AM, Li ZP, Lin HZ, Yang SQ. Cytokines and the pathogenesis of
non-alcoholic steatohepatitis. Gut 2005;54:303-6.
10. Chen J, Li X. Hypolipidemic effect of flavonoids from mulberry leaves
in triton WR-1339 induced hyperlipidemic mice. Asia Pac J Clin Nutr
2007;16 Suppl 1:290-4.
11. Butt MS, Nazir A, Sultan MT, Schroën K. Morus alba L. Nature’s
functional tonic. Trends Food Sci Technol 2008;19:505-12.
12. Soliman MM, Attia HF, El-Shazly SA, Saleh OM. Biomedical effects
of cinnamon extract on obesity and diabetes relevance in wistar rats.
Am J Biochem Mol Biol 2012;2:133-45.
13. Novelli EL, Diniz YS, Galhardi CM, Ebaid GM, Rodrigues HG,
Mani F, et al. Anthropometrical parameters and markers of obesity in
rats. Lab Anim 2007;41(1):111-9.
14. Sarikaphuti A, Nararatwanchai T, Hashiguchi T, Ito T, Thaworanunta S,
Kikuchi K, et al. Preventive effects of Morus alba L. anthocyanins on
diabetes in Zucker diabetic fatty rats. Exp Ther Med 2013;6(3):689-95.
15. Mbikay M. Therapeutic potential of Moringa oleifera leaves in chronic
hyperglycemia and dyslipidemia: A review. Front Pharmacol 2012;3:24.
16. Meiattini F, Prencipe L, Bardelli F, Giannini G, Tarli P. The
4-hydroxybenzoate/4-aminophenazone chromogenic system used
in the enzymic determination of serum cholesterol. Clin Chem
1978;24(12):2161-5.
17. Bucolo G, David H. Quantitative determination of serum triglycerides
by the use of enzymes. Clin Chem 1973;19(5):476-82.
18. Naito HK. HDL cholesterol. In: Kaplan A, editor. Clinical Chemistry:
Theory, Analysis and Correlation. St. Louis, Toronto, Princeton: C.V.
Mosby Co.; 1984. p. 1207-13, 437.
19. Trinder P. Determination of glucose in blood using glucose oxidase
with an alternative oxygen acceptor. Ann Clin Biochem 1969;6:24-33.
20. Wieland H, Seidel D. A simple specific method for precipitation of low
density lipoproteins. J Lipid Res 1983;24(7):904-9.
21. Eastham RD. Biochemical Values in Clinical Medicine. 7
ed. Bristol
England: John Wright & Sons Ltd.; 1985.
22. Satoh K. Serum lipid peroxide in cerebrovascular disorders determined
by a new colorimetric method. Clin Chim Acta 1978;90:37-43.
23. Montgoery HA, Dymock JF. The determination of nitrate in water.
Analyst 1961;86:414.
24. Young DS. Effect of Drugs on Clinical Laboratory Tests. 3
ed., Vol. 3.
Washington, DC: AACC Press; 1990. p. 6-12.
th
rd
25. Walterv M, Gerade H. A colorimetric method for determination
bilirubin in serum and plasma. Microchem J 1970;15:231.
26. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement
with the Folin phenol reagent. J Biol Chem 1951;193(1):265-75.
27. Matthews DR, Hosker JP, Rudenski AS, Naylor BA, Treacher DF,
Turner RC. Homeostasis model assessment: Insulin resistance and betacell
function
from fasting plasma glucose and insulin concentrations in

man.
Diabetologia 1985;28(7):412-9.
28. Banchroft JD, Stevens A, Turner DR. Theory and Practice of
Histological Techniques. 4
ed. New York, London, San Francisco,
Tokyo: Churchil Livingstone; 1996.
th
29. Abramoff MD, Magalhaes PJ, Ram SJ. Image processing with image J
Biophotonics Int 2004;11(7):36-42.
30. Lee YH, Pratley RE. The evolving role of inflammation in obesity and
the metabolic syndrome. Curr Diab Rep 2005;5(1):70-5.
31. Rayalam S, Della-Fera MA, Baile CA. Phytochemicals and regulation
of the adipocyte life cycle. J Nutr Biochem 2008;19(11):717-26.
32. Thielecke F, Boschmann M. The potential role of green tea catechins in
the prevention of the metabolic syndrome - A review. Phytochemistry
2009;70:11-24.
33. Rodrigues A, Pereira PC, Vicente AF, Brito JA, Bernardo MA,
Mesquita MF. Food intake, body mass index and body fat mass in
elderly. Asian J Clin Nutr 2012;4:107-15.
34. El-Beshbishy HA, Singab AN, Sinkkonen J, Pihlaja K. Hypolipidemic
and antioxidant effects of Morus alba L. (Egyptian mulberry) root
bark fractions supplementation in cholesterol-fed rats. Life Sci
2006;78(23):2724-33.
35. Wu ZH, Chen YQ, Zhao SP. Simvastatin inhibits ox-LDL-induced
inflammatory adipokines secretion via amelioration of ER stress in
3T3-L1 adipocyte. Biochem Biophys Res Commun 2013;432(2):365-9.
36. Sugimoto M, Arai H, Tamura Y, Murayama T, Khaengkhan P, Nishio T,
et al. Mulberry leaf ameliorates the expression profile of adipocytokines
by inhibiting oxidative stress in white adipose tissue in db/db mice.
Atherosclerosis 2009;204:388-94.
37. Koh KK, Quon MJ, Han SH, Lee Y, Kim, SJ, Park JP, et al. Differential
metabolic effects of pravastatin and simvastatin in hypercholesterolemic
patients. Atherosclerosis 2009;204(2):483-90.
38. Ballantyne CM, Olsson AG, Cook TJ, Mercuri MF, Pedersen TR,
Kjekshus J. Influence of low high-density lipoprotein cholesterol and
elevated triglyceride on coronary heart disease events and response to
simvastatin therapy in 4S. Circulation 2001;104(25):3046-51.
39. Son EL, Pal UK, Mandal PK, Hong GE, Kim SK, Lee CH.
Hypolipidaemic effect of processed sulfur, Allium tuberosum Rottl.
and fermented Allium tuberosum Rottl in rat. Asian J Anim Vet Adv
2012;7:812-21.
40. Fruchart JC, Brewer HB Jr, Leitersdorf E. Consensus for the use of
fibrates in the treatment of dyslipoproteinemia and coronary heart
disease. Fibrate Consensus Group. Am J Cardiol 1998;81:912-7.
41. Raveh O, Pinchuk I, Fainaru M, Lichtenberg D. Kinetics of lipid
peroxidation in mixtures of HDL and LDL, mutual effects. Free Radic
Biol Med 2001;31(11):1486-97.
42. Jain PG, Patil SD, Haswani NG, Girase MV, Surana SJ. Hypolipidemic
activity of Moringa oleifera Lam., Moringaceae, on high fat diet
induced hyperlipidemia in albino rats. Braz J Pharm 2010;20(6):969-73.
43. Lee J, Chae K, Ha J, Park BY, Lee HS, Jeong S, et al. Regulation of
obesity and lipid disorders by herbal extracts from Morus alba, Melissa
officinalis, and Artemisia capillaris in high-fat diet-induced obese
mice. J Ethnopharmacol 2008;115(2):263-70.
44. Venkatesan N, Devaraj SN, Devaraj H. Increased binding of LDL and
VLDL to apo B,E receptors of hepatic plasma membrane of rats treated
with Fibernat. Eur J Nutr 2003;42(5):262-71.
45. Matikainen N, Kahri J, Taskinen MR. Reviewing statin therapy in
diabetes – Towards the best practise. Prim Care Diabetes 2010;4(1):9-15.
46. Yao XM, Ye SD, Zai Z, Chen Y, Li XC, Yang GW, et al. Simvastatin
protects diabetic rats against kidney injury through the suppression
of renal matrix metalloproteinase-9 expression. J Endocrinol Invest
2010;33(2):292-6.
47. Sabri M, Ai J, Marsden PA, Macdonald RL. Simvastatin re-couples
dysfunctional endothelial nitric oxide synthase in experimental
subarachnoid hemorrhage. PLoS One 2011;6(2):e17062.
48. Delbosc S, Cristol JP, Descomps B, Mimran A, Jover B. Simvastatin
prevents angiotensin II-induced cardiac alteration and oxidative stress.
Hypertension 2002;40(2):142-7.
49. Galisteo M, Sánchez M, Vera R, González M, Anguera A, Duarte J,
et al. A diet supplemented with husks of Plantago ovata reduces the
development of endothelial dysfunction, hypertension, and obesity
by affecting adiponectin and TNF-alpha in obese Zucker rats. J Nutr
237
Asian J Pharm Clin Res, Vol 9, Suppl. 2, 2016, 231-238
Metwally et al.
2005;135(10):2399-404.
50. Ginsberg HN, Stalenhoef AF. The metabolic syndrome: Targeting
dyslipidaemia to reduce coronary risk. J Cardiovasc Risk
2003;10(2):121-8.
51. Hansawasdi C, Kawabata J. Alpha-glucosidase inhibitory
effect of mulberry (Morus alba) leaves on Caco-2. Fitoterapia
2006;77(7-8):568-73.
52. Li YG, Ji DF, Zhong S, Lv ZQ, Lin TB, Chen S. Hybrid of
1-deoxynojirimycin and polysaccharide from mulberry leaves treat
diabetes mellitus by activating PDX-1/insulin-1 signaling pathway
and regulating the expression of glucokinase, phosphoenolpyruvate
carboxykinase and glucose-6-phosphatase in alloxan-induced diabetic
mice. J Ethnopharmacol 2011;134(3):961-70.
53. Yada T, Nakata M, Shiraishi T, Kakei M. Inhibition by simvastatin, but
not pravastatin, of glucose-induced cytosolic Ca2 signalling and insulin
secretion due to blockade of L-type Ca2 channels in rat islet beta-cells.
Br J Pharmacol 1999;126(5):1205-13.
54. Mori N, Lee P, Yamamoto I, Nozawa S, Arai T. Insulin treatmentinduced
daily
changes
to
plasma
adiponectin
and
TNF-α
level
and
lipid

metabolism
parameters
in dogs
suffering
from
type
1
diabetes mellitus

Asian.
J
Anim
Vet
Adv
2011;6:844-50.
55. Lim HJ, Jin HG, Woo ER, Lee SK, Kim HP. The root barks of Morus
alba and the flavonoid constituents inhibit airway inflammation.
J Ethnopharmacol 2013;149(1):169-75.
56. Prasanna GS, Purnima A. Protective effect of leaf extract of Trichilia
connaroides on hypercholesterolemia induced oxidative stress. Int J
Pharmacol 2011;7:106-12.
57. Esposito LA, Melov S, Panov A, Cottrell BA, Wallace DC.
Mitochondrial disease in mouse results in increased oxidative stress.
Proc Natl Acad Sci U S A 1999;96:4820-5.
58. Radojkovic´ MM, Zekovic´ ZP, Vidovic´ SS, Kocˇar DD, Maškovic´
PZ. Free radical scavenging activity and total phenolic and flavonoid
contents of mulberry (Morus spp. L., Moraceae) extracts. Hemijska Ind
2012;66:547-52.
59. Wang Y, Xiang L, Wang C, Thang C, He Y. Antidiabetic and antioxidant
effects and phytochemicals of mulberry fruit (Morus alba L.)
polyphenol enhanced extract. PLoS One 2013;8:e71144.
60. O’Byrne DJ, Devaraj S, Grundy SM, Jialal I. Comparison of the
antioxidant effects of Concord grape juice flavonoids alpha-tocopherol
on markers of oxidative stress in healthy adults. Am J Clin Nutr
2002;76(6):1367-74.
61. Heeba G, Moselhy ME, Hassan M, Khalifa M, Gryglewski R, Malinski T.
Anti-atherogenic effect of statins: Role of nitric oxide, peroxynitrite
and haem oxygenase-1. Br J Pharmacol 2009;156:1256-66.
62. Choi JW, Pai SH, Kim SK, Ito M, Park CS, Cha YN. Increases in nitric
oxide concentrations correlate strongly with body fat in obese humans.
Clin Chem 2001;47(6):1106-9.
63. Avogaro A, de Kreutzenberg SV. Mechanisms of endothelial
dysfunction in obesity. Clin Chim Acta 2005;360(1-2):9-26.
64. Joost HG, Tschöp MH. NO to obesity: Does nitric oxide regulate fat
oxidation and insulin sensitivity? Endocrinology 2007;148:4545-7.
65. Wang Z, Nakayama T. Inflammation, a link between obesity and
cardiovascular disease. Mediators Inflamm 2010;2010:535918.
66. Fernández-Sánchez A, Madrigal-Santillán E, Bautista M, EsquivelSoto
J,
Morales-González A,
Esquivel-Chirino C,
et
al.
Inflammation,

oxidative
stress, and obesity.
Int J Mol Sci 2011;12(5):3117-32.
67. Codoñer-Franch P, Tavárez-Alonso S, Murria-Estal R, Megías-Vericat J,
Tortajada-Girbés M, Alonso-Iglesias E. Nitric oxide production is
increased in severely obese children and related to markers of oxidative
stress and inflammation. Atherosclerosis 2011;215(2):475-80.
68. Eo HJ, Park JH, Park GH, Lee MH, Lee JR, Koo JS, et al. Antiinflammatory
and
anti-cancer
activity
of
mulberry (Morus
alba
L.) root

bark.
BMC Complement Altern
Med 2014;14:200.
69. Choi EM, Hwang JK. Effects of Morus alba leaf extract on the
production of nitric oxide, prostaglandin E2 and cytokines in RAW264.7
macrophages. Fitoterapia 2005;76(7-8):608-13.
70. Iliodromitis EK, Andreadou I, Prokovas E, Zoga A, Farmakis D,
Fotopoulou T, et al. Simvastatin in contrast to postconditioning
reduces infarct size in hyperlipidemic rabbits: Possible role
of oxidative/nitrosative stress attenuation. Basic Res Cardiol
2010;105(2):193-203.
71. Piechota-Polanczyk A, Goraca A, Demyanets S, Mittlboeck M,
Domenig C, Neumayer C, et al. Simvastatin decreases free radicals
formation in the human abdominal aortic aneurysm wall via NF-?B.
Eur J Vasc Endovasc Surg 2012;44(2):133-7.
72. Radi ZA, Koza-Taylor PH, Bell RR, Obert LA, Runnels HA, Beebe JS,
et al. Increased serum enzyme levels associated with kupffer cell
reduction with no signs of hepatic or skeletal muscle injury. Am J
Pathol 2011;179:240-7.
73. Oh H, Ko EK, Jun JY, Oh MH, Park SU, Kang KH, et al. Hepatoprotective
and free radical scavenging activities of prenylflavonoids, coumarin,
and stilbene from Morus alba. Planta Med 2002;68(10):932-4.
74. Abbas AM, Sakr HF. Simvastatin and vitamin E effects on cardiac and
hepatic oxidative stress in rats fed on high fat diet. J Physiol Biochem
2013;69(4):737-50.
75. Cui B, Liu S, Lin X, Wang J, Li S, Wang Q, et al. Effects of Lycium
barbarum aqueous and ethanol extracts on high-fat-diet induced
oxidative stress in rat liver tissue. Molecules 2011;16(11):9116-28.
76. Doi K, Kojima T, Makino M, Kimura Y, Fujimoto Y. Studies on the
constituents of the leaves of Morus alba L. Chem Pharm Bull (Tokyo)
2001;49(2):151-3.
77. Ou TT, Kuo CY, Chyau CC, Lee HJ, Peng JS, Wang CJ. Improvement
of lipopolysaccharide-induced hepatic injuries and inflammation with
mulberry extracts. J Sci Food Agric 2013;93(8):1880-6.
Statistics
309 Views | 258 Downloads
Citatons
How to Cite
Ahmed, H. H., F. M. Metwally, H. Rashad, and A. M. Zaazaa. “INSIGHTS INTO THE ROLE OF MORUS ALBA IN REVERSING OBESITY-ASSOCIATED HEPATIC STEATOSIS AND RELATED METABOLIC DISORDER IN RATS”. Asian Journal of Pharmaceutical and Clinical Research, Vol. 9, no. 8, Oct. 2016, pp. 231-8, doi:10.22159/ajpcr.2016.v9s2.13674.
Section
Original Article(s)