, Volume 26, Issue 3, pp 769–778 | Cite as

Evaluation of the anti-inflammatory effect of an anti-platelet agent crinumin on carrageenan-induced paw oedema and granuloma tissue formation in rats

  • Uddipak Rai
  • Amrita Rawal
  • Sanjay Singh
Original Article


The present study was designed to explore the anti-inflammatory activity of an anti-platelet agent crinumin, by various in vitro and in vivo inflammation models. Firstly, crinumin protein was purified through cation exchange chromatography; then, in vitro activity was estimated by albumin denaturation assay and HRBC membrane stabilization assay. Carrageenan-induced paw oedema and cotton pellet-induced granuloma models were used for in vivo anti-inflammatory activity assessment in rats. In both models, rats were pre-treated for 7 days with crinumin (25–50 µg/ml) and diclofenac sodium (50 µg/ml). Expression of P-selectin (in serum and plasma) through ELISA and NF-κB (in paw and granulomatous tissues) through western blotting was checked. Our results showed that crinumin at both doses (25 or 50 µg/kg of b.w.) significantly (p < 0.05) reduced the paw oedema formation in a dose-dependent manner in the second phase of inflammation and significant (p < 0.05) reduction of wet and dry weight of granuloma was observed indicating the anti-inflammatory potential of crinumin. Crinumin decreased the expression of P-selectin and NF-κB indicating its potential role in decreasing platelet activation and healing inflammation. Histopathological studies additionally proved the efficacy of drug in treating inflammation. The results of the study suggest that the crinumin might have an inhibitory role in atherosclerosis as platelet aggregation and inflammation are the key processes involved in atherosclerotic disorders.


Crinumin Atherosclerosis P-selectin Carrageenan Platelet aggregation 



This study was financially supported by Institute Research Project grant from the Indian Institute of Technology (BHU), Varanasi, India (Number: IIT (BHU)/R&D/IRP/2015-16/2903/L). We would like to thank Dr. S.P. Singh, Professor in Department of Biochemistry, Banaras Hindu University for their support in providing research facilities.

Compliance with ethical standards

Conflict of interest

The authors declare that there are no conflicts of interest.


  1. Anbarasi A, Vidhya R (2015) Evaluation of in vitro anti-inflammatory activity of Tephrosia purpurea (Seed). Asian J Pharm Res 5:83–89CrossRefGoogle Scholar
  2. Borthakur A, Bhattacharyya S, Dudeja PK et al (2007) Carrageenan induces interleukin-8 production through distinct Bcl10 pathway in normal human colonic epithelial cells. Am J Physiol Gastrointest Liver Physiol 292:G829–G838CrossRefPubMedGoogle Scholar
  3. Bradford MM (1976) A rapid and sensitive for the quantitation of microgram quantitites of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefPubMedGoogle Scholar
  4. Burkill IH (1966) A dictionary of the economic products of the Malay peninsula. Ministry of Agriculture and Cooperative, Kuala LumpurGoogle Scholar
  5. Chaudhari MG, Joshi BB, Mistry KN (2013) In vitro anti-diabetic and anti-inflammatory activity of stem bark of Bauhinia purpurea. Bull Pharm Med Sci 1:139–150Google Scholar
  6. Chen M, Geng JG (2006) P-selectin mediates adhesion of leukocytes, platelets, and cancer cells in inflammation, thrombosis, and cancer growth and metastasis. Arch Immunol Ther Exp 54:75–84CrossRefGoogle Scholar
  7. Gawaz M, Langer H, May AE (2005) Platelets in inflammation and atherogenesis. J Clin Invest 115:3378–3384CrossRefPubMedPubMedCentralGoogle Scholar
  8. Ghani A (1998) Medicinal plants of Bangladesh: chemical constituents and uses. Asiatic Society of Bangladesh, DhakaGoogle Scholar
  9. Hussein SZ, Yusoff KM, Makpol S et al (2013) Gelam honey attenuates carrageenan-induced rat paw inflammation via NF-κB pathway. PLoS ONE 8:e72365CrossRefPubMedPubMedCentralGoogle Scholar
  10. Ioannis P, Constantinos G, Christos T et al (2014) NF-κB signaling at the crossroads of inflammation and atherogenesis: searching for new therapeutic links. Expert Opin Ther Targets 18:1089–1101CrossRefGoogle Scholar
  11. Karthik K, Bharath RKP, Venu PR et al (2013) Evaluation of anti-inflammatory activity of Canthium parviflorum by in vitro method. Indian J Res Pharm Biotechnol 1:729–730Google Scholar
  12. Kim YH, Kim KH, Han CS et al (2008) Anti-inflammatory activity of Crinum asiaticum Linne var. japonicum extract and its application as a cosmeceutical ingredient. J Cosmet Sci 59:419–430PubMedGoogle Scholar
  13. Leelaprakash G, Dass SM (2011) Invitro anti-inflammatory activity of methanol extract of Enicostemma axillare. Int J Drug Dev Res 3:189–196Google Scholar
  14. Libby P, Ridker PM, Maseri A (2002) Inflammation and atherosclerosis. Circulation 105:1135–1143CrossRefPubMedGoogle Scholar
  15. Mizushima Y, Kobayashi M (1968) Interaction of anti-inflammatory drugs with serum proteins, especially with some biologically active proteins. J Pharm Pharmacol 20:169–173CrossRefPubMedGoogle Scholar
  16. Müller K, Aichele S, Herkommer M et al (2010) Impact of inflammatory markers on platelet inhibition and cardiovascular outcome including stent thrombosis in patients with symptomatic coronary artery disease. Atherosclerosis 213:256–262CrossRefPubMedGoogle Scholar
  17. Okpo SO, Fatokun F, Adeyemi OO (2001) Analgesic and anti-inflammatory activity of Crinum glaucum aqueous extract. J Ethnopharmacol 78:207–211CrossRefPubMedGoogle Scholar
  18. Olajide OA, Makinde JM, Awe SO (1999) Effects of the aqueous extract of Bridelia ferruginea stem bark on carrageenan-induced oedema and granuloma tissue formation in rats and mice. J Ethnopharmacol 66:113–117CrossRefPubMedGoogle Scholar
  19. Ombrello C, Block RC, Morrell CN (2010) Our expanding view of platelet functions and its clinical implications. J Cardiovasc Transl Res 3:538–546CrossRefPubMedPubMedCentralGoogle Scholar
  20. Ridker PM (2001) High-sensitivity C-reactive protein potential adjunct for global risk assessment in the primary prevention of cardiovascular disease. Circulation 103:1813–1818CrossRefPubMedGoogle Scholar
  21. Shenoy S, Shwetha K, Prabhu K et al (2010) Evaluation of anti-inflammatory activity of Tephrosia purpurea in rats. Asian Pac J Trop Med 3:193–195CrossRefGoogle Scholar
  22. Singh KA, Kumar R, Rao GRK et al (2010) Crinumin, a chymotrypsin-like but glycosylated serine protease from Crinum asiaticum: purification and physicochemical characterisation. Food Chem 119:1352–1358CrossRefGoogle Scholar
  23. Singh KA, Nayak MK, Jagannadham MV et al (2011) Thrombolytic along with anti-platelet activity of crinumin, a protein constituent of Crinum asiaticum. Blood Cells Mol Dis 47:129–132CrossRefPubMedGoogle Scholar
  24. Steinhubl SR, Badimon JJ, Bhatt DL et al (2007) Clinical evidence for anti-inflammatory effects of antiplatelet therapy in patients with atherothrombotic disease. Vasc Med 12:113–122CrossRefPubMedGoogle Scholar
  25. Winter CA, Porter CC (1957) Effect of alterations in side chain upon anti-inflammatory and liver glycogen activities of hydrocortisone esters. J Am Pharm Assoc 46:515–519CrossRefGoogle Scholar
  26. Zhanhe J, Alan WM (2000) Amaryllidaceae. Science Press, Beijing, p 264Google Scholar

Copyright information

© Springer International Publishing AG 2017

Authors and Affiliations

  1. 1.Department of Pharmaceutical Engineering & TechnologyIndian Institute of Technology (BHU)VaranasiIndia

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