COMPARATIVE ANALYSIS OF BIOLOGICAL ACTIVITY OF SILYBUM MARIANUM L. FOOD SUPPLEMENTS AVAILABLE ON MARKET: INVITRO STUDY

  • Heba D. Hassanein
  • Mona M. Abdel Mohsen
  • Khaled A. Shams
  • Nahla S. Abdel-azim
  • Mahmoud A. Saleh
  • Nermine A. Ehsan
  • Faiza M. Hammouda Phytochemistry Dept., National Research Centre, 33 El Bohouth st. (former El Tahrir st.), Dokki, Giza, Egypt, P. O. 12622

Abstract

Objective: Silybum marianum L. Food Supplements that contain silymarin is widely used as a therapeutic agent in liver diseases. Many brands are available on the market in USA, Egypt, Europe and other countries. The objective of this study was to compare the biological activity in different preparations of silymarin available on the market in USA and Egypt using paracetamol-induced oxidative stress injury on primary cultured rat hepatocytes.

Methods: Forty four silymarin samples available on the market were collected from USA (24) and Egypt (20) and tested for hepat protective antioxidant effects on primary cultured rat hepatocytes. Cytotoxicity was measured by MTT [3-(4, 5-dimethyl-thiazol-2)-2,5-diphenyl tetrazolium bromide] assay and lactate dehydrogenase (LD) leakage into culture medium. Antioxidant effects were determined by glutathione reductase (GR), and Nitric oxide (NO) assays in silymarin, pretreated rat hepatocytes for 2 h followed by incubation with 25 mM paracetamol over a period of 1 h. Therapeutic index was calculated for each tested sample for comparative analysis.

Results: Silymarin preparations significantly decreased toxicity induced by paracetamol in rat hepatocytes, decreased lactate dehydrogenase leakage and prevented GSH depletion (P<0.01) and returned NO to basal levels in rat hepatocytes. The therapeutic index was 80, 40 and 20 for samples No. 20, 19 and 5 respectively.

Conclusions: The 44 different silymarin preparations tested in this study exhibited variation in antioxidant capacity and in reducing nitric oxide produced as a result of paracetamol injury. This variation in biological activity did not always correspond to the amount of silymarin recorded on samples.

 

Keywords: Silymarin, Primary Cultured Rat Hepatocytes, Antioxidant Effect, Glutathione Reductase, Nitric Oxide

Downloads

Download data is not yet available.

References

1. Kvasnicka F, Biba B, Sevcik R. Analysis of the active components of silymarin. J Chromatogr A 2003;990:239-45.
2. Corchete P. Silybum marianum (L.) Gaertn. The Source of Silymarin. In: Bioactive Molecules and Medicinal Plants. Ramawat KG, Merillon JM. (eds.); 2008. p. 123-48.
3. Radimer K, Bindewald B, Hughes J, Ervin B, Swanson C, Picciano MF. Dietary Supplement Use by US Adults: Data from the National Health and Nutrition Examination Survey, 1999–2000. Am J Epidemiol 2004;160:339-49.
4. Saller R, Meier R, Brignoli R. The use of silymarin in the treatment of liver diseases. Drugs 2001;61:2035-63.
5. Fried M, Navarro V, Afdhal N, Belle S, Wahed A, Hawke R, et al. Effect of silymarin (Milk Thistle) on liver disease in patients with chronic hepatitis C unsuccessfully treated with interferon therapy. JAMA 2012;308:274-82.
6. Patel N, Joseph C, Corcoran GB, Ray SD. Silymarin modulates doxorubicin-induced oxidative stress, Bcl-xL and p53 expression while preventing apoptotic and necrotic cell death in the liver. Toxicol Appl Pharmacol 2010;245:143-52.
7. Blumenthal M. The complete german commission E monographs: therapeutic guide to herbal medicines. Austin: American Botanical Council; 1998.
8. Mulrow C, Lawrence V, Jacobs B. Milk thistle: effects on liver disease and cirrhosis and clinical adverse effects. Evidence Report/Technology Assessment No. 21 (Contract 290-97-0012 to the San Antonio Evidence-based Practice Center, based at the University of Texas Health Science Center at San Antonio, and The Veterans Evidence-based Research, Dissemination, and Implementation Center, a Veterans Affairs Services Research and Development Center of Excellence). AHRQ Publication No. 01-E025. Rockville, Maryland: Agency for Healthcare Research and Quality; 2000.
9. Robinson M, Zhang X. The world medicines situation 2011. Traditional medicines: Global situation, issues and challenges. WHO, Geneva; 2011.
10. Yamamoto Y, Yamashita S. Plasma ratio of ubiquinol and ubiquinone as a marker of oxidative stress. Mol Aspects Med 1997;18:79-84.
11. Krinsky NI. Mechanism of action of biological antioxidants. Proc Soc Exp Biol Med 1992;200:248-54.
12. Baynes J, Dominiczak MH. Antioxidant defenses and reactive oxygen species. In: Medical Biochemistry, Ch. 11, Mosby. 2nd ed.; 1999. p. 133-7.
13. Burke AS, MacMillan-Crow LA, Hinson JA. Reactive nitrogen species in acetaminophen-induced mitochondrial damage and toxicity in mouse hepatocytes. Chem Res Toxicol 2010;23:1286–92.
14. Ni HM, Bockus A, Boggess N, Jaeschke H, Ding WX. Activation of autophagy protects against acetaminophen-induced hepatotoxicity. Hepatology 2012;55:222–32.
15. Yang X, Salminen WF. Kava extract, an herbal alternative for anxiety relief, potentiates acetaminophen-induced cytotoxicity in rat hepatic cells. Phytomedicine 2011;18:592–600.
16. Jaeschke H, Williams CD, McGill MR, Xie Y, Ramachandran A. Models of drug induced liver injury for evaluation of phytotherapeutics and other natural products. Food Chem Toxicol 2013;55:279–89.
17. Jaeschke H, McGill MR, C Williams D, Ramachandran A. Current issues with acetaminophen hepatotoxicity–a clinically relevant model to test the efficacy of natural products. Life Sci 2011;88:737–45.
18. Matsuura T, Kaibori M, Araki Y, Matsumiya M, Yamamoto Y, Ikeya Y, et al. Japanese herbal medicine, inchinkoto, inhibits inducible nitric oxide synthase induction in interlukin-1β-stimulated hepatocytes. Hepatol Res 2012;42:76-90.
19. Seglen PO. Preparation of isolated rat liver cells. Methods Cell Biol 1976;13:29-83.
20. Kiso Y, Tohkin M, Hikino H. Assay method forantihepatotoxic activity using galactosamine-induced cytotoxicity in primary-cultured hepatocytes. J Nat Prod 1983;46:841-7.
21. NIH Guide for the Care and Use of Laboratory Animals. NIH Publication: US Department of Health, Education and Welfare. Research Triangle Park, North Carolina; 1985.
22. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxic assay. J Immunol Methods 1983;65:263-71.
23. Carmichael J, DeGraff W, Gazdar A, Minna J, Mitchell J. Evaluation of tetrazolium based semiautomated colourimetric assay: assessment of chemosensetivity testing. Cancer Res 1987;47:936-42.
24. King J. The dehydrogenase of oxidoreductase-lactate dehydrogenase. In: Practical Clinical Enzymology. JC King. Van Nostrand London, UK; 1965. p. 83-93.
25. Goldberg DM, Spooner RJ. Glutathione reductase In: Bergmeyer HU, Bergmeyer J. GraBl M. (ed). Methods of enzymatic analysis. 3rd ed. Vol. 111. Verlag Chemie, Weinheim; 1983. p. 258-65.
26. Montgomery HAC, Dymock JF. The determination of nitrite in water. Analyst 1961;86:414–6.
27. Angulo P, Pate T, Jorgensen RA, Therneau TM, Lindor KD. Silymarin in the treatment of patients with primary biliary cirrhosis with a suboptimal response to ursodeoxycholicacid. Heaptol 2000;32:897-900.
28. Jacobs B, Dennehy C, Ramirez G, Sapp J, Lawrence VA. Milk thistle for the treatment of liver diseases. A systemic review and meta-analysis. Am J Med 2002;113:506-15.
29. Stickel F, Schuppan D. Herbal medicine in the treatment of liver diseases. Dig Liver Dis 2007;39:293-304.
30. Hikino H, Kiso Y, Wagner H, Fiebig M. Antihepatotoxic actions of flavonolignans from Silybum marianum fruits. Planta Med 1984;50:248-50.
31. Kidd P, Head C. A Review of the bioavailability and clinical efficacy of milk thistle phytosome: a silybin-phosphatidylcholine complex (Siliphos). Alternative Med Rev 2005;10:193-203.
32. Valenzuela A, Aspillaga M, Vial S, Guerra R. Selectivity of silymarin on the increase of the glutathione content in different tissues of the rat. Planta Med 1989;55:420-2.
33. Savio D, Harrasser PC, Basso G. Softgel capsule technology as an enhancer device for the absorption of natural principles in humans. A bioavailability cross-over randomized study on silybin. Arzneimittelforschung 1998;48:1104-6.
34. Manach C, Scalbert A, Morand C, Remesy C, Jimenez L. Polyphenols: food sources and bioavailability. Am J Clin Nutr 2004;79:727-47.
35. Flora K, Hahn M, Rosen H. Milk thistle (Silybum marianum) for the therapy of liver disease. Am J Gastroenterol 1998;93:139-43.
36. Koch HP, Ritschel WA. Bioavailability of silymarin, I: Volumes of distribution of silybin, silydianin, and silychristin from in-vitro data (author's transl). Arch Pharm 1981;314:515-7.
37. Feher J, Deak G, Muzes G. Liver-protective action of silymarin therapy in chronic alcoholic liver diseases. Orv Hetil 1989;130:2723-7.
38. Becker DE. Drug therapy in dental practice: general principles part 2-pharmacodynamic considerations. Anesthesia Progress 2007;54:19–24.
39. Muller PY, Milton MN. The determination and interpretation of the therapeutic index in drug development. Nat Rev Drug Discovery 2012;11:751-61.
Statistics
269 Views | 557 Downloads
How to Cite
Hassanein, H. D., M. M. A. Mohsen, K. A. Shams, N. S. Abdel-azim, M. A. Saleh, N. A. Ehsan, and F. M. Hammouda. “COMPARATIVE ANALYSIS OF BIOLOGICAL ACTIVITY OF SILYBUM MARIANUM L. FOOD SUPPLEMENTS AVAILABLE ON MARKET: INVITRO STUDY”. International Journal of Pharmacy and Pharmaceutical Sciences, Vol. 7, no. 11, Sept. 2015, pp. 55-60, https://innovareacademics.in/journals/index.php/ijpps/article/view/7958.
Section
Original Article(s)