Protective Effect of Proteins Derived from Seed Explant Cultures of Calotropis procera in a Preclinical Model of Arthritis

Abstract

Proteins isolated from the in vitro cultures of callus and roots developed from the germinating seed explants of Calotropis procera were evaluated for their protective effect in a rat model of arthritis at 1 and 5 mg/kg doses in two independent sets of experiments comprising of respective controls. Joint swelling, functional parameters, markers of inflammation, oxidative stress, tissue histology, and cyclooxygenase-2 expression in these rats were compared with those treated with the standard anti-inflammatory drug diclofenac on day 3. Both callus and root proteins produced a dose-dependent reduction in joint swelling, and at the 5-mg/kg dose, their inhibitory effect was more pronounced (57 and 55 % inhibition) than that of diclofenac (42 and 46 % in two sets). Both of the protein fractions alleviated functional limitations in arthritic rats and normalized the levels of prostaglandin E₂, tumor necrosis factor-α, and other biochemical markers of inflammation and oxidative stress, maintained tissue architecture, and suppressed cyclooxygenase-2 expression compared to arthritic controls. This study demonstrates that the proteins derived from the in vitro culture of C. procera explants have potential in the treatment of arthritis.

Key words

Calotropis procera - Apocynaceae - callus - inflammation - medicinal plant - roots

• References

• 1 Gad HA, El-Ahmady SH, Abou-Shoer MI, Al-Aziz MM. Application of chemometrics in authentication of herbal medicines: a review. Phytochem Anal 2013; 24: 1-24
• 2 Hedberg I. Botanical methods in ethnopharmacology and the need for conservation of medicinal plants. J Ethnopharmacol 1993; 38: 121-128
• 3 Canter PH, Thomas H, Ernst E. Bringing medicinal plants into cultivation: opportunities and challenges for biotechnology. Trends Biotechnol 2005; 23: 180-185
• 4 Bourgaud F, Gravot A, Milesi S, Gontier E. Production of plant secondary metabolites: a historical perspective. Plant Sci 2001; 161: 839-851
• 5 Kumar VL, Arya S. Medicinal uses and pharmacological properties of Calotropis procera . In: Govil JN, editor Recent progress in medicinal plants, Vol. 11. Houston, TX: Studium Press; 2006: 373-388
• 6 Kumar VL, Basu N. Anti-inflammatory activity of latex of Calotropis procera . J Ethnopharmacol 1994; 44: 123-125
• 7 Dewan S, Sangraula H, Kumar VL. Preliminary studies on the analgesic activity of latex of Calotropis procera . J Ethnopharmacol 2000; 73: 307-311
• 8 Alencar NM, Oliveira JS, Mesquita RO, Lima MW, Vale MR, Etchells JP, Freitas CD, Ramos MV. Pro- and anti-inflammatory activities of the latex from Calotropis procera (Ait) R.Br. are triggered by compounds fractionated by dialysis. Inflamm Res 2006; 55: 559-564
• 9 Soares PM, Lima SR, Matos SG, Andrade MM, Patrocínio MC, de Freitas CD, Ramos MV, Criddle DN, Cardi BA, Carvalho KM, Assreuy AM, Vasconcelos SM. Antinociceptive activity of Calotropis procera latex in mice. J Ethnopharmacol 2005; 99: 125-129
• 10 Teixeira FM, Ramos MV, Soares AA, Oliveira RSB, Almeida-Filho LCP, Oliveira JS, Marinho-Filho JDB, Carvalho CPS. In vitro tissue culture of the medicinal shrub Calotropis procera to produce pharmacologically active proteins from plant latex. Process Biochem 2011; 46: 1118-1124
• 11 Kumar VL, Sharma N, da Costa Souza IC, Ramos MV, da Silveira Carvalho CP. Proteins derived from in vitro culture of the callus and roots of Calotropis procera ameliorate acute inflammation in the rat paw. Appl Biochem Biotechnol 2015; 175: 1724-1731
• 12 Crofford LJ. Use of NSAIDs in treating arthritis. Arthritis Res Ther 2013; 15 (Suppl. 03) S2
• 13 Kumar VL, Chaudhary P, Ramos MV, Mohan M, Matos MPV. Protective effect of proteins derived from the latex of Calotropis procera against inflammatory hyperalgesia in monoarthritic rats. Phytother Res 2011; 25: 1336-1341
• 14 Kumar VL, Roy S. Protective effect of latex of Calotropis procera in Freundʼs complete adjuvant induced monoarthritis. Phytother Res 2009; 23: 1-5
• 15 Kumar VL, Chaudhary P, Oliveira RM, Ramos MV. Calotropis procera latex proteins ameliorate functional limitations associated with adjuvant induced inflammation in rat. Musculoskelet Biol 2014; 1: 1
• 16 Lau D, Baldus S. Myeloperoxidase and its contributory role in inflammatory vascular disease. Pharmacol Ther 2006; 111: 16-26
• 17 Salvemini D, Wang ZQ, Wyatt PS, Bourdon DM, Marino MH, Manning PT, Currie MG. Nitric oxide: a key mediator with early and late phase of carrageenan-induced rat paw inflammation. Br J Pharmacol 1996; 118: 829-838
• 18 Kunsch C, Sikorski JA, Sundell CL. Oxidative stress and the use of antioxidants for the treatment of rheumatoid arthritis. Cur Med Chem Immunol Endocr Metab Agents 2005; 5: 249-258
• 19 Choy EH, Panayi GS. Cytokine pathways and joint inflammation in rheumatoid arthritis. N Engl J Med 2001; 344: 907-916
• 20 Gan TJ. Diclofenac: an update on its mechanism of action and safety profile. Curr Med Res Opin 2010; 26: 1715-1731
• 21 Guruprasad B, Chaudhary P, Choedon T, Kumar VL. Artesunate ameliorates functional limitations in Freundʼs complete adjuvant-induced monoarthritis in rat by maintaining oxidative homeostasis and inhibiting COX-2 expression. Inflammation 2015; 38: 1028-1035
• 22 Ellman GL. Tissue sulfhydryl groups. Arch Biochem Biophys 1959; 82: 70-77
• 23 Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 1979; 95: 351-358
• 24 Ikeda U, Kanbe T, Shimada K. Adrenomedullin increases inducible nitric oxide synthase in rat vascular smooth muscle cells stimulated with interleukin-1. Hypertension 1996; 27: 1240-1244
• 25 Bradley PP, Priebat DA, Christensen RD, Rothstein G. Measurement of cutaneous inflammation: estimation of neutrophil content with an enzyme marker. J Invest Dermatol 1982; 78: 206-209