A Comparative Study upon the Therapeutic Indices of Some Natural and Synthetic Anti-inflammatory Agents

Document Type : Original Article


1 Department of Immunology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran

2 Department of Pharmacology, School of Medicine, Toxicology Research Centre, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran

3 Department of Pharmacognosy, School of Pharmacy, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran


The aim of the present study was to compare the therapeutic indices of several agents used in treatment of inflammatory conditions which included: Vitamin E, hydro-alcoholic extracts of Glycyrrhiza glabra, Matricaria aurea, dexamethasone, piroxicam and diclofenac using Wehi-164 fibrosarcoma cells.
Materials and Methods
Cytotoxicity evaluation was based on vital dye exclusion assay. Matrix-metalloproteinases inhibition (MMPI) was assessed by gelatinase zymography method. The collected data were used to estimate the IC50 (50% MMPI concentration), LC50 (50% cytotoxicity concentration) and the therapeutic index (LC50/IC50).
Among the natural anti-inflammatory agents used, M. aurea was the least toxic and the most effective inhibitor of MMP. Vitamin E not only increased MMP activity, but also was the most toxic of all the agents tested. Next in terms of toxicity to vitamin E was G. glabra. Diclofenac was more toxic than both piroxicam and dexamethasone.
Findings from this study suggest that medicinal plants reputed to have anti-inflammatory properties are not equally effective and safe. In order to assess the implications of these findings, further in vitro and in vivo studies are needed.


1. DiMartino MJ, High W, Galloway WA, Crimmin MJ. Preclinical antiartherric activity of matrix metalloproteinase inhibitors. Ann N Y Acad Sci 1994; 732:413-413.
2. Janssens S, Lijinen HR. What has been learned about the cardiovascular effects of matrix metalloproteinases from moue models? Cardiovasc Res 2006; 69:585-594.
3. Egelblad M, Werb Z. New functions for matrix metalloproteinases in cancer progression. Nat Rev Cancer 2002; 2:161-174.
4. Johnson C, Sung HJ, Lessner SM, Fini ME, Galis ZS. Matrix metalloproteinase-9 is required for adequate angiogenic revascularization of ischemic tissues: potential role in capillary branching. Circ Res 2004; 94:262¬268.
5. Barrett AJ, Rawlings ND, Woessnar JF. Handbook of Proteolytic Enzymes. Academic Press; 2004.
6. Woessner JF Nagase H. I nhibition of the MMPs. Textbook of Matrix Metalloproteinases and TIMPs. Oxford University Press; 2000.
7. Chirco R, Liu XW, Jung KK, Kim HR. Novel functions of TIMPs in cell signaling. Cancer Metastasis Rev 2006; 25:99-113.
8. Whittaker M, Floyd CD, Brown P, Gearing AJ. Design and therapeutic application of matrix metalloproteinase inhibitors. Chem Rev 1999; 99:2735-2776.
9. Smalley WE, Ray WA, Daugherty JR, Griffin MR. Nonsteroidal anti-inflammatory drugs and the incidence of hospitalizations for peptic ulcer disease in elderly persons. Am J Epidemiol 1995; 141:539-545.
10. Tipton DA, Lyle B, Babich H, Dabbous MKh. In vitro cytotoxicity and anti-inflammatory effects of myrrh oil on human gingival fibroblasts and epithelial cells. In Vitro Toxicol 2003; 17:301-310
11. Kimura I, Yoshikawa M, Kobayashi S, Sugihara Y, Suzuki M, Oominami H, et al. New triterpenes, myrrhanol A and myrrhanone A, from guggulgum resin and their potent anti-inflammatory effect on adjuvant-induced air pouch granuloma of mice. Bioorg Med Chem Lett 2001; 11:985-989.
12. Sheela ML, Ramakrishna MK, Salimath BP. Angiogenic and proliferative effects of the cytokine VEGF in Ehrlich ascites tumor cells is inhibited by Glycyrrhiza glabra. Int Immunopharmacol 2006; 6:494-498.
13. Rajesh MG, Latha MS. Protective activity of Glycyrrhiza glabra Linn. on carbon tetrachloride-induced peroxidative damage. Indian J Pharmacol 2004; 36:284-287.
14. Al-Ismail KM, Aburjai T. Antioxidant activity of water and alcohol extracts of chamomile flowers, anise seeds and dill seeds. J Sci Food Agric 2004; 84:173-178.
15. Abu-Rabia A. Palestinian plant medicines for treating renal disorders: An inventory and brief history. Alter Complem Ther? 2005; 11:295-300.
16. Orbe J, Rodn'guez JA, Arias R, Belzunce M, Nespereira B, Pe'rez-Ilzarbe M, et al. Antioxidant vitamins increase the collagen content and reduce MMP-1 in a porcine model of atherosclerosis: implications for plaque stabilization. Atherosclerosis 2003; 167:45-53.
17. Mirshafiey A, Khodadadi A, Rehmy BH, Khorramizadeh MR, Eslami MR, Razavi A, et al. Sodium alginate as a novel therapeutic option in experimental colitis. Scand J Immunol 2005; 61:316-321.
18. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 1983; 65:53-63.
19. Williamson EM, Okapako DT, Evans FJ. Pharmacological Methods in phytotherapy research, volume 1: Selection, preparation and pharmacological evaluation of plant material. John Willey and Sons; Chichester? 1989.p.15-23.
20. Heussen C, Dowdle EB. Electrophoretic analysis of plasminogen activator in polyacrylamide gels containing sodium dodecyl sulfate and copolymerized substrates. Ann Biochem 1980; 102:196-202.
21. Saunders FR, Hughes A, Heather M, Wallace HM. Does polyamine depletion influence NSAID-induced cytotoxicity in colorectal cancer? Abstracts Toxicol 2006; 226:12-77.
22. Tomisato W, Tanaka K, Katsu T, Kakuta H, Sasaki K, Tsutsumi S, et al. Membrane permeabilization by non- steroidal anti-inflammatory drugs. Biochem Biophys Res Commun 2004; 323: 1032-1039.
23. Overall CM, Lopez-Otin C. Strategies for inhibition of cancer: Innovation for the post-trial era. Nat Rev Cancer 2002; 2: 657-672.
24. Parks WC, Wilson CL, Lopez-Boado YS. Matrix metalloproteinases as modulators of inflammation and innate immunity. Nat Rev Immunol 2004; 4:617-629
25. Traber M, Atkinson J. Vitamin E: antioxidant and nothing more. Free Radic Biol Med 2007; 43:4-15.
26. Azzi A. Molecular mechanism of a-tocopherol action. Free Radic Biol Med 2007; 43: 16-21.
27. Chakraborti S, Mandal A, Das S, Chakrabort Ti. Inhibition of Na+/Ca2+ exchanger by peroxynitrite in microsomes of pulmonary smooth muscle: role of matrix metalloproteinase-2. Biochim Biophys Acta 2004; 1671:70- 78.
28. Slack RS. The effect of vitamin A and E on neuroblastoma growth and differentiation. Thesis submitted in partial fulfilment of the requirements for the degree of Doctorate of Philosophy in Biochemistry. Ottawa, Canada, 1991.
29. Theriault A, Chao JT, Wang Q, Gapor A, Adeli K. Tocotrienol: a review of its therapeutic potential. Clin Biochem 1999; 32: 309-319.
30. Kline K, Yu W, Sanders BG.Vitamin E: Mechanisms of action as tumor cell growth inhibitors. J Nutr 2001; 131:161S-163S.
31. Fotakis G, Timbrell JA. In vitro cytotoxicity assays: Comparison of LDH, neutral red, MTT and protein assay in hepatoma cell lines following exposure to cadmium chloride. Toxicol Lett 2006; 160:171-177.
32. Shariftabrizi A, Nifli AP, Ansari M, Saadat F, Ebrahimkhani MR, Alizadeh N, et al. Matrix metalloproteinase 2 secretion in WEHI 164 fibrosarcoma cells is nitric oxide-related and modified by morphine. Eur J Pharmacol 2006; 530:33-39.
33. Romero D, Gomez-Zapata M, Luna A, Garcia-Fernandez AJ.Morphological characterisation of BGM (Buffalo Green Monkey) cell line exposed to low doses of cadmium chloride. Toxicol In Vitro 2003; 17:293-299.