, Volume 20, Issue 1, pp 27–37 | Cite as

Lupeol, a triterpenoid isolated from Calotropis gigantea latex ameliorates the primary and secondary complications of FCA induced adjuvant disease in experimental rats

  • Venkatesan Saratha
  • Sorimuthu Pillai Subramanian
Research Article


Rheumatoid arthritis (RA) is a chronic inflammatory autoimmune disorder that affects 1% of the adult population worldwide. Calotropis gigantea is a xerophytic, latex producing medicinal plant widely distributed in nature. Different parts of the plant have been traditionally used for the treatment of various ailments including arthritis. In the present study, we have isolated and characterized lupeol, a pentacyclic triterpene from the dialyzable fraction of the latex and evaluated the anti-arthritic properties of lupeol in Freund’s Complete Adjuvant (FCA) induced arthritis in rats. Lupeol (50 mg/kg b.w/day) was administered orally to AA rats for 4 weeks. The alterations in body weight gain, paw volume, RBC, WBC, Hb, EPO, ESR, platelets and PCV were recorded. The activities of serum AST, ALT and ALP were also assayed. The levels of lipid profile were estimated. The levels of pro-inflammatory cytokines as well as anti-inflammatory cytokines such as TNF-α, IL-1β, IL-6 and IL-10 were also analyzed. The results of present study indicate the anti-inflammatory and anti-arthritic activity of lupeol present in the C. gigantea latex.


Calotropis gigantea latex Lupeol adjuvant induced arthritis Hyperlipidemia Pro-inflammatory cytokines 

Supplementary material

10787_2011_95_MOESM1_ESM.pdf (24 kb)
Supplementary material 1 (PDF 24 kb)


  1. Agarwal RB, Rangar VD (2003) Antiinflammatory and antiarthritic activities of lupeol and 19a–H lupeol isolated from Strobilanthus callosus and Strobilanthus ixiocephala roots. Indian J Pharmacol 35:384–387Google Scholar
  2. Alarcón GS (1995) Epidemiology of rheumatoid arthritis. Rheum Dis Clin North Am 21:589–604PubMedGoogle Scholar
  3. Arend WP, Dayer JM (1995) Inhibition of the production and effects of interleukin-1 and tumor necrosis factor alpha in rheumatoid arthritis. Arthritis Rheum 38:151–160PubMedCrossRefGoogle Scholar
  4. Auger I, Lepecuchel L, Roudier J (2002) Interaction between heat-shock protein 73 and HLA-DRB1 alleles associated or not with rheumatoid arthritis. Arthritis Rheum 46:929–933PubMedCrossRefGoogle Scholar
  5. Aworh OC, Kasche V, Apampa O (1994) Purification and some properties of sodom-apple latex proteinases. Food Chem 50:359–362CrossRefGoogle Scholar
  6. Baigent C, Keech A, Kearney PM, Blackwell L, Buck G, Pollicino C (2005) Efficacy and safety of cholesterol-lowering treatment: prospective meta-analysis of data from 90,056 participants in 14 randomised trials of statins. Lancet 366:1267–1278PubMedCrossRefGoogle Scholar
  7. Bani S, Kaul A, Khan B, Ahmad SF, Suri KA, Gupta BD, Satti NK, Qazi GN (2006) Suppression of T lymphocyte activity by lupeol isolated from Crataeva religiosa. Phytother Res 20:279–287PubMedCrossRefGoogle Scholar
  8. Bentley DP, Cavill I, Ricketts C, Peake S (1979) A method for the investigation of reticulo endothelial iron kinetics in man. Br J Haematol 43:619–624PubMedCrossRefGoogle Scholar
  9. Billingham ME (1983) Models of arthritis and the search for anti-arthritic drugs. Pharmacol Ther 21:389–428PubMedCrossRefGoogle Scholar
  10. Bondurant MC, Koury MJ (1986) Anaemia induces accumulation of erythropoietin mRNA in the kidney and liver. Mol Cell Biol 6:2731–2733PubMedGoogle Scholar
  11. Burton W, Morrison A, Maclean A, Ruderman E (2006) Systematic review of studies of productivity loss due to rheumatoid arthritis. Occup Med (Lond) 56:18–27CrossRefGoogle Scholar
  12. Chamundeeswari D, Vasantha J, Gopalakrishnan S, Sukumar E (2003) Free radical scavenging activity of the alcoholic extract of Trewia polycarpa roots in arthritic rats. J Ethnopharmacol 88:51–56PubMedCrossRefGoogle Scholar
  13. Choy EH, Panayi GS (2001) Cytokine pathways and joint inflammation in rheumatoid arthritis. N Engl J Med 344:907–916PubMedCrossRefGoogle Scholar
  14. Collins R, Armitage J, Parish S, Sleigh P, Peto R (2003) MRC/BHF heart protection study of cholesterol-lowering with simvastatin in 5,963 people with diabetes: a randomised placebo-controlled trial. Lancet 361:2005–2016PubMedCrossRefGoogle Scholar
  15. Ding Y, Nguyen HT, Kim SI, Kim HW, Kim YH (2009) The regulation of inflammatory cytokine secretion in macrophage cell line by the chemical constituents of Rhus sylvestris. Bioorg Med Chem Lett 19:3607–3610PubMedCrossRefGoogle Scholar
  16. Domsalla A, Melzig MF (2008) Occurrence and properties of proteases in plant lattices. Planta Med 74:699–711PubMedCrossRefGoogle Scholar
  17. Falholt K, Lund B, Falholt W (1973) An easy colorimetric micromethod for routine determination of free fatty acids in plasma. Clin Chim Acta 46:105–111PubMedCrossRefGoogle Scholar
  18. Fitzpatrick LR, Green C, Frauenhoffer EE, French KJ, Zhuang Y, Maines LW, Upson JJ, Paul E, Donahue H, Mosher TJ, Smith CD (2011) Attenuation of arthritis in rodents by a novel orally-available inhibitor of sphingosine kinase. Inflammopharmacology 19:75–87PubMedCrossRefGoogle Scholar
  19. Fournier C (2005) Where do T cells stand in rheumatoid arthritis? Jt Bone Spine 72:527–532CrossRefGoogle Scholar
  20. Fries JF, Singh G, Lenert L, Furst DE (1990) Aspirin, hydroxychloroquine, and hepatic enzyme abnormalities with methotrexate in rheumatoid arthritis. Arthritis Rheum 33:1611–1619PubMedCrossRefGoogle Scholar
  21. Fulgentius N, Lugemwa FU, Huang Y, Michael Bentley D, Michael J, Mendel A (1990) A Heliothis zea antifeedant from the abundant birchbark triterpene betulin. J Agric Food Chem 38:493–496CrossRefGoogle Scholar
  22. Furst DE, Eystone ECK, Fleischmann R, Mease P, Breedveld FC, Smolen JS (2010) Updated consensus statement on biological agents for the treatment of rheumatic diseases 2009. Ann Rheum Dis 69:2–29CrossRefGoogle Scholar
  23. Geetha T, Varalakshmi P (1999) Effect of lupeol and lupeol linoleate on lysosomal enzymes and collagen in adjuvant-induced arthritis in rats. Mol Cell Biochem 201:83–87PubMedCrossRefGoogle Scholar
  24. Geetha T, Varalakshmi P (2001) Anti-inflammatory activity of lupeol and lupeol linoleate in rats. J Ethnopharmacol 76:77–80PubMedCrossRefGoogle Scholar
  25. Geetha T, Varalakshmi P, Latha RM (1998) Effect of triterpenes from Crataeva nurvala stem bark on lipid peroxidation in adjuvant induced arthritis in rats. Pharmacol 37:191–195Google Scholar
  26. Georgiadis AN, Voulgari PV, Argyropoulou MI, Alamanos Y, Elisaf M, Tselepis AD et al (2008) Early treatment reduces the cardiovascular risk factors in newly diagnosed rheumatoid arthritis patients. Semin Arthritis Rheum 38:13–29PubMedCrossRefGoogle Scholar
  27. Gidez LI, Miller GJ, Burstein M, Slagle S, Eder HA (1982) Separation and quantitation of subclasses of human plasma high density lipoproteins by a simple precipitation procedure. J Lipid Res 1206–1223Google Scholar
  28. Gutiérrez JM, Gutierrez Ricart C, Ricart W, Castiñeira MJ, Vendrell J, Richart C (1999) Plasma levels of the soluble fraction of tumor necrosis factor receptors 1 and 2 are independent determinants of plasma cholesterol and LDL-cholesterol concentrations in healthy subjects. Atherosclerosis 146:321–327PubMedCrossRefGoogle Scholar
  29. Hisadome M, Fukuda T, Sumichika H, Hanano T, Adachi KA (2000) A novel anti-rheumatic drug suppresses tumor necrosis factor-alpha and augments interleukin-10 in adjuvant arthritic rats. Eur J Pharmacol 409:331–335PubMedCrossRefGoogle Scholar
  30. Imam S, Azhar I, Hasan MM, Ali MS, Ahmed SW (2007) Two triterpenes lupanone and lupeol isolated and identified from Tamarindus indica linn. Pak J Pharm Sci 20:125–127PubMedGoogle Scholar
  31. Jeekmann WE, Fandrey J, Freede S, Pagel H (1994) Inhibition of erythropoietin production by cytokines, implications for the anaemia involved in inflammatory states. Ann NY Acad Sci 718:300–309CrossRefGoogle Scholar
  32. Kataoka H, Horiyamac S, Yamaki M, Oku H, Ishiguro K, Katagi T (2002) Anti-inflammatory and anti-allergic activities of hydroxylamine and related compounds. Biol Pharm Bull 25:1436–1441PubMedCrossRefGoogle Scholar
  33. Kawakami M, Murase T, Itakura H, Yamada N, Ohsawa N, Takaku F (1986) Lipid metabolism in endotoxic rats: decrease in hepatic triglyceride lipase activity. Microbiol Immunol 30:849–854PubMedGoogle Scholar
  34. King J (1965) The phosphohydrolases-acid and alkaline phosphatases. In: Van D (ed) Practical Clinical Enzymology. Nostrand Company Limited, London, pp 191–208Google Scholar
  35. Lam FF, Wong HH, Ng ES (2004) Time course and substance P effects on the vascular and morphological changes in adjuvant-induced monoarthritic rats. Int Immunopharmacol 4:299–310PubMedCrossRefGoogle Scholar
  36. Lee DM, Weinblatt ME (2001) Rheumatoid arthritis. Lancet 358:903–911PubMedCrossRefGoogle Scholar
  37. Lemeshko VV, Brovkovich VM, Listopad AP (1991) A simple method for estimation of phospholipids in biological objects. Ukr Biokhim Zh 63:60–66PubMedGoogle Scholar
  38. Liz-Grana M, Gomez-Reino Carnota JJ (2005) Tumour necrosis factor. Genetics, cell action mechanism and involvement in inflammation. Allergol Immunol Clin 16:140–149Google Scholar
  39. Lucetti DL, Lucetti EC, Bandeira MA, Veras HN, Silva AH, Leal LK (2010) Anti-inflammatory effects and possible mechanism of action of lupeol acetate isolated from Himatanthus drasticus (Mart.) Plumel. J Inflamm (Lond) 7:60CrossRefGoogle Scholar
  40. Maria M, Engeniusz M, Miroslaw K, Maria K, Iwona P (1983) Adjuvant induced disease in rats. Clinical findings and morphological and biochemical changes in blood and histological changes in internal organs. Rheumatologia 21:213–245Google Scholar
  41. Mathis S, Jala VR, Haribabu B (2007) Role of leukotriene B4 receptors in rheumatoid arthritis. Autoimmun Rev 7:12–17PubMedCrossRefGoogle Scholar
  42. Means RT, Krantz SB (1996) Inhibition of human erythroid colony-forming units by interferons alpha and beta: differing mechanisms despite shared receptor. Exp Hematol 24:204–208PubMedGoogle Scholar
  43. Megraw RE, Dunn DE, Biggs HG (1979) Manual and continuous-flow colorimetry of triacylglycerols by a fully enzymic method. Clin Chem 25:273–278PubMedGoogle Scholar
  44. Mosser DM (2003) The many faces of macrophage activation. J Leukoc Biol 73:209–212PubMedCrossRefGoogle Scholar
  45. Neves C, Jorge R, Barcelos A (2009) The network of methotrexate toxicity. Acta Reumatol Port 34:11–34PubMedGoogle Scholar
  46. Nguemfo EL, Dimo T, Dongmo AB, Azebaze AG, Alaoui K, Asongalem AE, Cherrah Y, Kamtchouing P (2009) Anti-oxidative and anti-inflammatory activities of some isolated constituents from the stem bark of Allanblackia monticola Staner L.C (Guttiferae). Inflammopharmacol 17:37–41CrossRefGoogle Scholar
  47. Nielsen OJ, Andersen LS, Ludwigsen E, Bouchelouche P, Hansen TM, Birgens H (1990) Anaemia of rheumatoid arthritis: serum erythropoietin concentrations and red cell distribution width in relation to iron status. Ann Rheum Dis 49:349–353PubMedCrossRefGoogle Scholar
  48. Olhagen B (1965) On the aetiopathogenesis of rheumatoid arthritis. Ann Clin Res 7:119–128Google Scholar
  49. Pahwa R, Chatterjee VC (1998) The toxicity of Indian Calotropis procera RBr latex in the black rat, Rattus rattus Linn. Vet Hum Toxicol 30:305–308Google Scholar
  50. Parekh AC, Jung DH (1970) Cholesterol determination with ferric acetate–uranium acetate reagent and sulphuric acid ferrous sulphate reagent. Anal Chem 42:1423–1427CrossRefGoogle Scholar
  51. Pearson CM (1963) Experimental joint disease observations on adjuvant-induced arthritis. J Chronic Dis 16:863–874PubMedCrossRefGoogle Scholar
  52. Preetha SP, Kanniappan M, Selvakumar E, Nagaraj M, Varalakshmi P (2006) Lupeol ameliorates aflatoxin B1-induced peroxidative hepatic damage in rats. Comp Biochem Physiol C Toxicol Pharmacol 143:333–339PubMedCrossRefGoogle Scholar
  53. Rahman MA, Wilcock CC (1991) A taxonomic revision of Calotropis (Asclepiadaceae). Nord J Bot 11:301–408CrossRefGoogle Scholar
  54. Rainsford KD (1982) Adjuvant polyarthritis in rats: is this a satisfactory model for screening anti-arthritic drugs. Agents Actions 12:452–458PubMedCrossRefGoogle Scholar
  55. Ramos MV, Aguiar VC, Silva Xavier AA, Lima MW, Bandeira GP, Etchells JP (2006) Latex proteins from the plant Calotropis procera are partially digested upon in vitro enzymatic action and are not immunologically detected in faecal material. Fitoterapia 77:251–256PubMedCrossRefGoogle Scholar
  56. Rehman Q, Lane NE (2001) Therapeutic approaches for preventing bone loss in inflammatory arthritis. Arthritis Res 3:221–227PubMedCrossRefGoogle Scholar
  57. Reinhart WH (1992) The influence of iron deficiency on erythrocyte deformability. Br J Haematol 80:550–555PubMedCrossRefGoogle Scholar
  58. Saleem M (2009) Lupeol, a novel anti-inflammatory and anti-cancer dietary triterpenes. Cancer Lett 285:109–115PubMedCrossRefGoogle Scholar
  59. Saratha V, Subramanian S, Sivakumar S (2010) Evaluation of wound healing potential of Calotropis gigantea latex studied on excision wounds in experimental rats. Med Chem Res 19:936–947CrossRefGoogle Scholar
  60. Schopf L, Savinainen A, Anderson K, Kujawa J, DuPont M, Silva M, Siebert E, Chandra S, Morgan J, Gangurde P, Wen D, Lane J, Xu Y, Hepperle M, Harriman G, Ocain T, Jaffee B (2006) IKKbeta inhibition protects against bone and cartilage destruction in a rat model of rheumatoid arthritis. Arthritis Rheum 54:3163–3173PubMedCrossRefGoogle Scholar
  61. Shedlofsky SL, Swim AT, Robinson JM, Gallicchio VS, Cohen DA, McClain CJ (1987) Interleukin-1 (IL-1) depresses cytochrome P450 levels and activities in mice. Life Sci 40:2331–2336PubMedCrossRefGoogle Scholar
  62. Sholichin M, Yamasaki K, Kasai R, Tanaka O (1980) 13C Nuclear Magnetic Resonance of Lupane type triterpenes, Lupeol, Betulin and Betulinic acid. Chem Pharm Bull 28:1006–1008CrossRefGoogle Scholar
  63. Simon LS (2000) DMARDs in the treatment of rheumatoid arthritis: current agents and future developments. Int J Clin Pract 54:243–249PubMedGoogle Scholar
  64. Singh H, Kumar S, Dewan S, Kumar VL (2000) Inflammation induced by latex of Calotropis procera—a new model to evaluate anti-inflammatory drugs. J Pharmacol Toxicol Methods 43:219–224PubMedCrossRefGoogle Scholar
  65. Skogh T, Gustafsson D, Kjellberg M, Husberg M (2003) Twenty eight joint count disease activity score in recent onset rheumatoid arthritis using C reactive protein instead of erythrocyte sedimentation rate. Ann Rheum Dis 62:681–682PubMedCrossRefGoogle Scholar
  66. Sokka T (2003) Work disability in early rheumatoid arthritis. Clin Exp Rheumatol 21:71–74Google Scholar
  67. Stoerk HC, Bielinski TC, Budzilovich T (1954) Chronic polyarthritis in rats injected with spleen in adjuvant. Am J Pathol 30:616–621Google Scholar
  68. Subramanian S, Saratha V (2010) Evaluation of antibacterial activity of Calotropis gigantea latex extract on selected pathogenic bacteria. J Pharm Res 3:517–552Google Scholar
  69. Van Doornum S, McColl G, Wicks IP (2004) Atorvastatin reduces arterial stiffness in patients with rheumatoid arthritis. Ann Rheum Dis 63:1571–1575PubMedCrossRefGoogle Scholar
  70. Van Doornum S, Brand C, King B, Sundararajan V (2006) Increased case fatality rates following a first acute cardiovascular event in patients with rheumatoid arthritis. Arthritis Rheum 54:2061–2068PubMedCrossRefGoogle Scholar
  71. Wang Y, Fang Y, Huang W, Zhou X, Wang M, Zhong B (2005) Effect of sinomenine on cytokine expression of macrophages and synoviocytes in adjuvant arthritis rats. J Ethnopharmacol 98:37–43PubMedCrossRefGoogle Scholar
  72. Weber J, Werre JM, Julius HW, Marx JJ (1988) Decreased iron absorption in patients with active rheumatoid arthritis, with and without iron deficiency. Ann Rheum Dis 47:404–409PubMedCrossRefGoogle Scholar
  73. White WB, West CR, Borer JS, Gorelick PB, Lavange L, Pan SX (2007) Risk of cardiovascular events in patients receiving celecoxib: a meta-analysis of randomized clinical trials. Am J Cardiol 99:91–98PubMedCrossRefGoogle Scholar
  74. William Carey M, Rao NV, Kumar BR, Mohan GK (2010) Anti-inflammatory and analgesic activities of methanolic extract of Kigelia pinnata DC flower. J Ethnopharmacol 130:79–82CrossRefGoogle Scholar
  75. Williams RO (1998) Rodent models of arthritis: relevance for human disease. Clin Exp Immunol 114:330–332PubMedCrossRefGoogle Scholar
  76. Winder CV, Lembke LA, Stephens MD (1969) Comparative bioassay of drugs in adjuvant-induced arthritis in rats: flufenamic acid, mefenamic acid, and phenylbutazone. Arthritis Rheum 12:472–482PubMedCrossRefGoogle Scholar
  77. Winter CA, Risley EA, Nuss CW (1962) Carageenan induced edema in hind paw of the rats as an assay for anti-inflammatory drugs. Proc Soc Exp Bio Med 111:544–547Google Scholar

Copyright information

© Springer Basel AG 2011

Authors and Affiliations

  • Venkatesan Saratha
    • 1
  • Sorimuthu Pillai Subramanian
    • 1
  1. 1.Department of BiochemistryUniversity of MadrasChennaiIndia

Personalised recommendations