Free Radical Scavenging and Antioxidant effects of Tolfenamic Acid in L-NAME-Induced Hypertensive Rats


Published: 2023-12-09

DOI: 10.56557/upjoz/2023/v44i233799

Page: 392-400

R. Dhakshinamoorthi

Department of Biochemistry and Biotechnology, Cardiovascular Biology Laboratory, Annamalai University, Annamalai Nagar-608 002, Tamil Nadu, India.

B. Raja *

Department of Biochemistry and Biotechnology, Cardiovascular Biology Laboratory, Annamalai University, Annamalai Nagar-608 002, Tamil Nadu, India.

K. Ragul

Department of Biochemistry and Biotechnology, Cardiovascular Biology Laboratory, Annamalai University, Annamalai Nagar-608 002, Tamil Nadu, India.

*Author to whom correspondence should be addressed.


Objectives: The aim of this study was to determine the antioxidant and free radical scavenging properties of tolfenamic acid (TA) against N-Nitro-L-arginine methyl ester hydrochloride (L-NAME) induced hypertension.

Materials and Methods: The rats were divided into five groups at random: Group I (control rats), Group II (control TA), Group III (L-NAME), Group IV (L-NAME+TA), and Group V (L-NAME+Enalapril). For four weeks, rats were given L-NAME (40mg/kg body weight) dissolved in drinking water to induce hypertension. Intraperitoneal injections of TA (50mg/kg body weight) and enalapril (0.7 mg/kg body weight) were given to rats.

Results: The results showed that the In vitro free radical scavenging effect of TA on DPPH and ABTS was concentration dependent. In vivo studies with L-NAME resulted in increases in blood pressure, lipid hydroperoxides (LOOH), and thiobarbituric acid reactive substances (TBARS). In addition, the level of the non-enzymatic antioxidant reduced glutathione (GSH) and other enzyme antioxidant activities in the heart and aorta, including superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (Gpx), are reduced. The level of nitric oxide metabolism in the erythrocyte aorta of L-NAME rats was increased.

Conclusions: These findings imply that tolfenamic acid acts as an antihypertensive and antioxidant agent in L-NAME-induced hypertension.

Keywords: Tolfenamic acid, L-NAME, hypertension, free radicals, antioxidants

How to Cite

Dhakshinamoorthi, R., Raja, B., & Ragul, K. (2023). Free Radical Scavenging and Antioxidant effects of Tolfenamic Acid in L-NAME-Induced Hypertensive Rats. UTTAR PRADESH JOURNAL OF ZOOLOGY, 44(23), 392–400.


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Jagannathan R, Patel SA, Ali MK, Narayan KV. Global updates on cardiovascular disease mortality trends and attribution of traditional risk factors. Current diabetes reports. 2019;19:1-12.

Ahmad KA, Yuan Yuan D, Nawaz W, Ze H, Zhuo CX, Talal B, Qilong D. Antioxidant therapy for management of oxidative stress induced hypertension. Free radical research. 2017;51(4):428-438.

Förstermann U. Nitric oxide and oxidative stress in vascular disease. Pflügers Archiv-European Journal of Physiology. 2010;459: 923-939.

Wilcox CS. Oxidative stress and nitric oxide deficiency in the kidney: A critical link to hypertension? American Journal of Physiology-Regulatory, Integrative and Comparative Physiology. 2005;289(4): R913-R935.

Hakami WES. Determination and Environmental Remediation of Non-Steroidal Anti-Inflammatory Drugs from Model and Real Samples.

Salekin S, Sharmin T, Rahman MS. Synthesis, Characterization and Analgesic Activity of Cadmium (II) Complex of Tolfenamic Acid: Synthesis, Characterization and Analgesic Activity. Dhaka University Journal of Pharmaceutical Sciences. 2020;19(1):59-64.

Saravanakumar M, Raja B. Veratric acid, a phenolic acid attenuates blood pressure and oxidative stress in L-NAME induced hypertensive rats. European Journal of Pharmacology. 2011;671(1-3):87-94.

Ghasemi K, Ghasemi Y, Ebrahimzadeh MA. Antioxidant activity, phenol and flavonoid contents of 13 citrus species peels and tissues. Pak J Pharm Sci. 2009; 22(3):277-281.

Liu D, Shi J, Ibarra AC, Kakuda Y, Xue SJ. The scavenging capacity and synergistic effects of lycopene, vitamin E, vitamin C, and β-carotene mixtures on the DPPH free radical. LWT-Food Science and Technology. 2008;41(7):1344-1349.

Nampoothiri LP, Agarwal A, Gupta S. Effect of co-exposure to lead and cadmium on antioxidant status in rat ovarian granulose cells. Archives of toxicology. 2007;81:145-150.

Sinha AK. Colorimetric assay of catalase. Analytical biochemistry. 1972;47(2):389-394.

Rotruck JT, Pope AL, Ganther HE, Swanson AB, Hafeman DG, Hoekstra W. Selenium: Biochemical role as a component of glutathione peroxidase. Science. 1973;179(4073):588-590.

Boxer M, Ellman L, Geller R, Wang CA. Anemia in primary hyperparathyroidism. Archives of Internal Medicine. 1977;137(5): 588-590.

Lowry OH, Rosebrough NJ, Farr AL. Parasites of the common crow (Corvusbrachyrhynchos Brehm, 1822) in insular newfoundland1. Biol Chem. 2011; 193:265-275.

Niehaus Jr WG, Samuelsson B. Formation of malonaldehyde from phospholipid arachidonate during microsomal lipid peroxidation. European journal of biochemistry. 1968;6(1):126-130.

Jiang ZY, Hunt JV, Wolff SP. Ferrous ion oxidation in the presence of xylenol orange for detection of lipid hydroperoxide in low density lipoprotein. Analytical biochemistry. 1992;202(2):384-389.

Majithiya JB, Paramar AN, Balaraman R. Pioglitazone, a PPARγ agonist, restores endothelial function in aorta of streptozotocin-induced diabetic rats. Cardiovascular Research. 2005;66(1):150-161.

Gulcin İ. Antioxidants and antioxidant methods: An updated overview. Archives of toxicology. 2020;94(3):651-715.

Büyükokuroğlu ME, Gülçin I, Oktay M, Küfrevioğlu OI. In vitro antioxidant properties of dantrolene sodium. Pharmacological Research. 2001;44(6): 491-494.

Ionita P. The chemistry of DPPH· free radical and congeners. Int. J. Mol. Sci. 2021;22:1545.

Gulcin I, Mshvildadze V, Gepdiremen A, Elias R. Screening of antioxidant and antiradical activity of monodesmosides and crude extract from Leontice smirnowii Tuber. Phytomedicine. 2006;13: 343–351.

Altay A, Tohma H, Durmaz L, Taslimi P, Korkmaz M, Gulcin I, Koksal E. Preliminary phytochemical analysis and evaluation of in vitro antioxidant, antiproliferative, antidiabetic and anticholinergics effects of endemic Gypsophila taxa from Turkey. J. Food Biochem. 2019;43:e12908.

Munteanu IG, Apetrei C. Analytical methods used in determining antioxidant activity: A review. Int. J. Mol. Sci. 2021;22: 3380.

Balaydın HT, Gulcin I, Menzek A, Goksu S, Sahin E. Synthesis and antioxidant properties of diphenylmethane derivative bromophenols including a natural product. J. Enzym. Inhib. Med. Chem. 2010;25: 685–695.

Kedare SB, Sing RP. Genesis and development of DPPH method of antioxidant assay. J. Food Sci. Technol. 2011;48:412–422.

Cetinkaya Y, Gocer H, Menzek A, Gulcin I. Synthesis and antioxidant properties of (3,4-dihydroxyphenyl) (2,3,4-trihydroxyphenyl) methanone and its derivatives. Arch. Pharm. 2012;345:323–33.

Koksal E, Bursal E, Dikici E, Tozoglu F, Gulcin I. Antioxidant activity of Melissa officinalis leaves. J. Med. Plants Res. 2011;5:217–222.

Ilyasov IR, Beloborodov VL, Selivanova IA. Three ABTS•+ radical cation-based approaches for the evaluation of antioxidant activity: Fast-and slow-reacting antioxidant behavior. Chemical Papers. 2018;72:1917-1925.

Husain N, Kumar A, Radicals F. Reactive oxygen species and natural antioxidants: A review. Advances in Bioresearch. 2012; 3(4):164-175.

Afzal F, Khurshid R, Ashraf M, Kazi AG. Reactive oxygen species and antioxidants in response to pathogens and wounding. In Oxidative damage to plants. Academic Press. 2014;397-424.

Wadhwa N, Mathew BB, Jatawa S, Tiwari A. Lipid peroxidation: mechanism, models and significance. Int J Curr Sci. 2012;3(1); 29-38.

Sarban S, Kocyigit A, Yazar M, Isikan UE. Plasma total antioxidant capacity, lipid peroxidation, and erythrocyte antioxidant enzyme activities in patients with rheumatoid arthritis and osteoarthritis. Clinical biochemistry. 2005;38(11):981-986.

Piacenza L, Zeida A, Trujillo M, Radi R. The superoxide radical switch in the biology of nitric oxide and peroxynitrite. Physiological Reviews. 2022;102(4):1881-1906.