Oriental Pharmacy and Experimental Medicine

, Volume 16, Issue 3, pp 217–224 | Cite as

Antioxidant, genoprotective and immunomodulatory potential of Vitex negundo leaves in experimental arthritis

  • Harsha Lad
  • Ankita Joshi
  • Deepti Dixit
  • Harsha Sharma
  • Deepak BhatnagarEmail author
Research Article


Vitex negundo is a medicinal plant used in Indian and folklore medicines to cure various ailments including arthritis. In the present study, the hydroethanolic extract of V. negundo leaves (VNE) were evaluated for antioxidant, genoprotective and immunomodulatory activity in Freund’s complete adjuvant (FCA) induced arthritis. VNE was evaluated for in vitro antioxidant activity using various parameters including plasmid nicking assay. VNE (200 mg/kg body weight) was orally administered to FCA induced arthritic rats to evaluate its genoprotective effects on peripheral blood leukocytes using the alkaline comet assay. Furthermore, the effects of VNE treatment on serum proinflammatory mediators such as tumor necrosis factor-α (TNF-α) and interleukin -1α (IL-1α) and the hematological parameters such as total RBC count, hematocrit and hemoglobin were measured in arthritic rats. The results showed that VNE exhibited potential in vitro antioxidant and DNA protecting activities in a concentration dependent manner. The phytoconstituents of VNE showed a strong and positive correlation with the antioxidant properties. DNA damage in the peripheral blood leukocytes of arthritic animals was significantly decreased after VNE treatment. VNE showed inhibition of serum proinflammatory cytokines (TNF-α and IL-1α) however, the inhibition was statistically not-significant. The arthritic animals showed significant increase in hemolysis, which was restored by VNE treatment. The results of the present study indicate that VNE may be a source of antioxidant, genoprotective with immunomodulatory activity, which may be attributed to its antioxidant phytoconstituents.


Vitex negundo Antioxidant Plasmid Freund’s adjuvant Comet assay TNF-α IL-1α 


Compliance with ethical standards

Ethical Statement

All the experiments were ap proved and conducted as per the guidelines of the Institutional Animal Ethics Committee (IAEC) (No. Biochem /01/2013–14).

Conflict of Interest

The authors declare that there are no conflicts of interest.


  1. Altindag O, Karakoc M, et al. (2007) Increased DNA damage and oxidative stress in patients with rheumatoid arthritis. Clin Biochem 40:167–171CrossRefPubMedGoogle Scholar
  2. Bashir S, Harris G, et al. (1993) Oxidative DNA damage and cellular sensitivity to oxidative stress in human autoimmune diseases. Ann Rheum Dis 52:659–666CrossRefPubMedPubMedCentralGoogle Scholar
  3. Chapple IL (1997) C reactive oxygen species and antioxidants in inflammatory diseases. J Clin Periodontol 24:287–296CrossRefPubMedGoogle Scholar
  4. Chattopadhyay P, Hazarika S, et al. (2012) Vitex negundo inhibits cyclooxygenase-2 inflammatory cytokine mediated inflammation on carageneen induced rat hind paw edema. Pharmacog Res 4:134–137CrossRefGoogle Scholar
  5. De Vizcaya-Ruiz A, Rivero-Muller A, et al. (2003) Hematotoxicity response in rats by the novel copper-based anticancer agent: casiopeina II. Toxicology 194:103–113CrossRefPubMedGoogle Scholar
  6. Devi PR, Kumari SK, et al. (2007) Effect of Vitex negundo leaf extract on the free radicals scavengers in complete Freund's adjuvant induced arthritic rats. Ind J Clin Biochem 22:143–147CrossRefGoogle Scholar
  7. Dharmasiri MG, Jayakody JRAC, et al. (2003) Anti-inflammatory and analgesic activities of mature fresh leaves of Vitex negundo. J Ethnopharmacol 87:199–206CrossRefPubMedGoogle Scholar
  8. Dixit D, Dixit AK, et al. (2013) Radioprotective effect of Terminalia chebula Retzius extract against γ-irradiation-induced oxidative stress. Biomed Aging Pathol 3:83–88CrossRefGoogle Scholar
  9. Droge W (2002) Free radicals in the physiological control of cell function. Review. Physiol Rev 82:47–95CrossRefPubMedGoogle Scholar
  10. Gandhi NM, Nair CKK (2005) Radiation protection by Terminalia chebula: some mechanistic aspects. Mol Cell Biochem 277:43–48CrossRefPubMedGoogle Scholar
  11. Garcia O, Romero I, et al. (2007) Measurement of DNA damage on silver stained comets using free Internate software. Mutat Res 627:186–190CrossRefPubMedGoogle Scholar
  12. Geckil H, Ates B, et al. (2005) Antioxidant, free radical scavenging and metal chelating characteristics of propolis. Am J Biochem Biotech 1:27–31CrossRefGoogle Scholar
  13. Hajizadeh S, DeGroot J, et al. (2003) TeKoppele JM, Tarkowski a, Collins LV. Extracellular mitochondrial DNA and oxidatively damaged DNA in synovial fluid of patients with rheumatoid arthritis. Arthritis Res Ther 5:234–240CrossRefGoogle Scholar
  14. Halliwell B, Gutteridge JMC (1985) Chronic inflammation and autoimmune disease. In: free radicals in biology and medicine, clarendon press, 279Ioannides C (2000) xenobiotic metabolism and bioactivation by cytochromes P450. In: Wiseman H, Goldfarb P, Ridgway T, Wiseman A (eds) Biomolecular free radical toxicity: causes and prevention. Chichester, John Wiley and Sons Limited, p. 103Google Scholar
  15. Halliwell B, Gutteridge JMC (1999) Free radicals in biology and medicine, 3rd edn. Oxford University Press, OxfordGoogle Scholar
  16. Hua BL, Chi CW, et al. (2008) Antioxidant properties in vitro and total phenolic contents in methanol extracts from medicinal plants. LWT 41:385–390CrossRefGoogle Scholar
  17. Karaman A, Binici DN, et al. (2011) Comet assay and analysis of micronucleus formation in patients with rheumatoid arthritis. Mutat Res 721:1–5CrossRefPubMedGoogle Scholar
  18. Kavitha C, Ramesh M, et al. (2012) Toxicity of Moringa oleifera seed extract on some hematological and biochemical profiles in a freshwater fish, Cyprinus carpio. Experiment Toxicol Pathol 64:681–687CrossRefGoogle Scholar
  19. Kumar PP, Kumaravel S, et al. (2010) Screening of antioxidant activity, total phenolics and GC-MS study of Vitex negundo. Afr J Biochem Res 4:191–195Google Scholar
  20. Lad H, Dixit D, et al. (2015) Antioxidant and antiinflammatory effects of Vitex negundo on Freund’s complete adjuvant induced arthritis. Int J Pharma Pharmaceutical Sci 7:81–85Google Scholar
  21. Lakshmanashetty RH, Nagaraj VB, et al. (2010) In vitro antioxidant activity of Vitex negundo L. Leaf Extracts Chiang Mai J Sci 37(3):489–497Google Scholar
  22. Laughton MJ, Halliwell B, et al. (1987) Antioxidant and prooxidant actions of the plant phenolics quercetin, gossypol and myricetin. Biochem Pharmacol 36:717–720CrossRefGoogle Scholar
  23. Lee JC, Kim HR, et al. (2002) Antioxidant property of an ethanol extract of the stem of Opuntia Ficus – indica Var. Saboten J Agri Food Chem 50:6490–6649CrossRefGoogle Scholar
  24. Mehta AB, Hoffbrand AV (1999) Haematological aspects of systemic disease. In: Hoffbrand AV, Catovsky D, Tuddenham EGD (eds) Postgraduate Haematology; Chapt 59, 5th edn. Blackwell Publishing Ltd, HobokenGoogle Scholar
  25. Mukinda JT, Syce JA (2007) Acute and chronic toxicity of the aqueous extract of Artemisia Afra in rodents. J Ethnopharmacol 112:138–144CrossRefPubMedGoogle Scholar
  26. Mustafa RA, Azizah AH, et al. (2010) Total phenolic compounds, flavonoids and radical scavenging activity of selected tropical plants. J Food Sci 75:C28–C35CrossRefPubMedGoogle Scholar
  27. Negri G, Teixeira EW (2011) Hydroxycinnamic acid amide derivatives, phenolic compounds and antioxidant activities of extracts of pollen samples from Southeast Brazil. J Agric Food Chem 59:5516–5522CrossRefPubMedGoogle Scholar
  28. Panday GS, Chunekar KC (1998) Bhav prakash nighantu Varanasi: Chaukhambha Bharati Academy p. 344–345Google Scholar
  29. Pandey A, Bani S, et al. (2012) Anti-arthritic activity of agnuside mediated through the down-regulation of inflammatory mediators and cytokines. Inflamm Res 61:293–304CrossRefPubMedGoogle Scholar
  30. Ryan KA, Smith MF Jr, et al. (2004) Reactive oxygen and nitrogen species differentially regulate toll-like receptor 4-mediated activation of NF kappa B and interleukin-8 expression. Infect Immun 72:2123–2130CrossRefPubMedPubMedCentralGoogle Scholar
  31. Sabnis M (2006) Chemistry and pharmacology of ayurvedic medicinal plants, 1st edn. Varanasi: Chaukambha Amarbharati Prakashan; p. 363–366Google Scholar
  32. Shahidi F (2000) Antioxidants in food and food antioxidants. Nahrung. 44:158–163CrossRefPubMedGoogle Scholar
  33. Singh H, Dixit A, et al. (2015) Comparative evaluation of total phenolic content, total flavonoid content and DPPH free radical scavenging activity of different plant parts of Vitex negundo. Int J Pharm Pharm Sci 7:144–147Google Scholar
  34. Tandon VR (2005) Medicinal uses and biological activities of Vitex negundo. Nat Pro Rad 4:162–165Google Scholar
  35. Telang RS, Chatterjee S, et al. (1999) Studies on analgesic and anti-inflammatory activities of Vitex negundo Linn. Ind J Pharmacol 31:363–366Google Scholar
  36. Valko M, Leibfritz D, et al. (2007) Free radicals and antioxidants in normal physiological functions and human disease. Int J Biochem Cell Biol 39:44–84CrossRefPubMedGoogle Scholar
  37. Venkatesha SH, Berman BM, et al. (2011) Herbal medicinal products target defined biochemical and molecular mediators of inflammatory autoimmune arthritis. Bioorg Med Chem 19:21–29CrossRefPubMedGoogle Scholar
  38. Wong C, Li H, et al. (2006) A systematic survey of antioxidant activity of 30 Chinese medicinal plants using the ferric reducing antioxidant power assay. Food Chem 97:705–711CrossRefGoogle Scholar
  39. Yadav AS, Bhatnagar D (2007a) Modulatory effect of spice extracts on iron induced lipid peroxidation in rat liver. Biofactors 29:147–157CrossRefPubMedGoogle Scholar
  40. Yadav AS, Bhatnagar D (2007b) Free radical scavenging activity, metal chelation and antioxidant power of some of the Indian spices. Biofactors 31:219–227CrossRefPubMedGoogle Scholar
  41. Yizhong C, Qiong L, et al. (2004) Antioxidant activity and phenolic compounds of 112 traditional Chinese medicinal plants associated with anticancer. Life Sci 74:2157–2184CrossRefGoogle Scholar
  42. Zargar M, Azizah AH, et al. (2011) Bioactive compounds and antioxidant activity of different extracts from Vitex negundo leaf. J Med Plants Res 5:2525–2532Google Scholar
  43. Zheng CJ, Zhao XX, et al. (2014) Therapeutic effects of standardized Vitex negundo seeds extract on complete Freund's adjuvant induced arthritis in rats. Phytomedicine 21:838–846CrossRefPubMedGoogle Scholar

Copyright information

© Institute of Korean Medicine, Kyung Hee University and Springer Science+Business Media Dordrecht 2016

Authors and Affiliations

  • Harsha Lad
    • 1
  • Ankita Joshi
    • 1
  • Deepti Dixit
    • 1
  • Harsha Sharma
    • 1
  • Deepak Bhatnagar
    • 1
    Email author
  1. 1.School of BiochemistryDevi Ahilya UniversityIndoreIndia

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