Anti-immunosuppressive Effects of Chromolaena odorata (Lf.) King & Robinson (Asteraceae) Leaf Extract in Cyclophosphamide-injected Balb/C Mice
Leonora P. Nudo1,3 and Elena S. Catap, Ph.D.1,2*
1Natural Sciences Research Institute,
University of the Philippines,Diliman, Quezon City
2Institute of Biology, College of Sciences, University of the Philippines,
Diliman, Quezon City
3Department of Science, School of Sciences and Engineering,
University of Asia and the Pacific, Ortigas Center, Pasig City
Chromolaena odorata (Lf.) King & Robinson leaf extract was evaluated for its immunomodulating activity in Balb/C mice by employing a number of immune response assays namely macrophage phagocytic activity, splenocyte proliferation, production of reactive oxygen species (ROS), and plasma lysozyme levels. The optimum concentration of C. odorata was determined to be 100 µg/mL through in vitro assays. Subsequently, three (3) groups of 12 mice each were used in in vivo experiments, namely: (1) cyclophosphamide-induced (30mg/kg body weight) immunosuppressed mice (Cy- injected or positive control); (2) C. odorata extract + Cy-treated mice (C group); and (3) phosphate-buffered saline (PBS)-injected mice as the negative control group. When compared with PBS-treated mice, the Cy-treated mice showed significantly lower phagocytic activity, cellular proliferation, production of reactive oxygen species (ROS), and plasma lysozyme levels. Significant improvement in macrophage phagocytic activity and cellular proliferation was exhibited by the plant extract-treated mice when compared with Cy-injected mice only. The extract from C. odorata also improved superoxide production and plasma lysozyme activity compared with the Cy-injected mice. These results demonstrated the immunopotentiating activities of the C. odorata leaf extract on the innate immunity of Balb/C mice. Also, the extract could potentially reverse a drug-induced immunosuppression as confirmed through in vivo experiments. Indeed, there is a great potential of the plant to be utilized as source of biologically active products and metabolites for drug development.
Immunomodulation through stimulation or suppression may help in maintaining disease-free state (Ghule et al. 2006). Herbal medicine has become an integral part of standard healthcare, based on a combination of time-honoured traditional usage and ongoing scientific research (Tiwari et al. 2004). Since the ancient systems of medicine, plants have been extensively used as a source of medicine to promote health and to maintain the body’s resistance against infection by potentiating immunity, re-establishing body equilibrium and . . . . . . . . . . . . . . . .
AJAYA KUMAR R, SRIDEVI K, VIJAYA KUMAR N, NADURI S, RAJAGOPAL S. 2004. Anticancer and immunostimulating compounds from Androraphis paniculata. J Ethnopharm 92: 291-295.
BAFNA AR, MISHRA SH. 2006. Immunostimulatory effect of methanol extract of Cucurligo orchioides on immunosuppressed mice. J Ethnopharm 104: 1-4.
BIN-HAFEEZ B, AHMAD I, HAQUE R, RAISUDDIN S. 2001. Protective effect of Cassia occidentalis L. on cyclophosphamide-induced suppression of humoral immunity in mice. J Ethnopharm 75: 13-18.
BORREGAARD N. 1985. The respiratory burst of phagocytosis: Biochemistry and subcellular localization. Immunol Lett 11: 165-171.
CHOI EM, KOO JK, HWANG J-K. 2004. Immune cell stimulating activity of mucopolyssacharide isolated from yam (Dioscorea batatasi). J Ethnopharm 91: 1-6.
GHULE BV, MURUGANANTHAN G, NAKHAT PD, YEOLE PG. 2006. Immunostimulant effects of Capparis zeylanica Linn. leaves. J Ethnopharm 108: 311-315.
GREENBERG S, GRINSTEIN S. 2002. Phagocytosis and innate immunity. Curr Opin Immunol 14: 136-145.
JI Z, TANG Q, ZHANG Z, YANG Y, JIA W, PAN Y. 2007. Immunomodulation of RAW 264.7 macrophages by GLIS, a proteopolysaccharide from Ganoderma lucidum. J Ethnopharm 112: 445-450.
LING SK, PISAR MM, MAN S. 2007. Platelet-activating factor (PAF) receptor binding antagonist activity of the methanol extracts and isolated flavonoids from Chromolaena odorata (L.) King and Robinson. Biol Pharm Bull 30(6): 1150-52.
LUNARDI F, LIMA JHC, ASSREUY J. 2006. Comparative study of respiratory burst induced by phorbol ester and zymosan in human granulocytes. Clin Biochem 39: 78-85.
MANOSROI A, SARAPHANCHOTIWITTHAYA A, MANOSROI J. 2005. In vitro immunomodulatory effect of Pouteria cambodiana (Pierre ex Dubard) Baehni extract. J Ethnopharm 101: 90-94.
MOSSMAN T. 1983. Rapid colorimetric assay for cell cellular growth and survival: application to proliferation and cytotoxicity assays. J Ethnopharm 65: 55-63.
NGONO NGANE A, EBELLE ETAME R, NDIFOR F, BIYITI L, AMVAM ZOLLO PH, BOUCHET P. 2006. Antifungal activity of Chromolaena odorata (L.) King & Robinson (Asteraceae) of Cameroon. Chemotherapy 52: 103-106.
OWOYELE VB, ADEDIJI JO, SALODAYE AO. 2005. Anti-inflammatory activity of aqueous leaf extract of Chromolaena odorata. Inflammopharmacology 13(5- 6): 479-484.
PHANTT, HUGHESMA, CHERRYGW. 1998. Enhanced proliferation of fibroblasts and endothelial cells treated with an extract of the leaves of Chromolaena odorata (Eupolin), an herbal remedy for treating wounds. Plast Reconstr Surg 101(3): 756-765.
PHAN TT, ALLEN J, HUGHES MA, CHERRY G, WOJNAROWSKA F. 2000. Upregulation of adhesion complex proteins and fibronectin by human keratinocytes treated with an aqueous extract from the leaves of Chromolaena odorata (Eupolin). Eur J Dermatol 10(7): 522.
PHAN TT, HUGHES MA, CHERRY GW. 2001a. Effects of an aqueous extract from the leaves of Chromolaena odorata (Eupolin) on the proliferation of human keratinocytes and on their migration in an in vitro model of reepithelialization. Wound Repair Regen 9(4): 305-313.
PHAN TT, WANG L, SEE P, GRAYER RJ, CHAN SY, LEE ST. 2001b. Phenolic compounds of Chromolaena odorata protect cultured skin cells from oxidative damage: implication for cutaneous wound healing. Biol Pharm Bull 24(12): 1373-79.
SECOMBES CJ. 1998. Fish Immunology Workshop (Workshop Manual). Australia: University of Tasmania.
SUKSAMRARN A, CHOTIPONG A, SUAVANSRI T, BOONGIRD S, TIMSUKSAI P, VIMUTTIPONG S, CHUAYNUGUL A. 2004. Antimycobacterial activity and cytotoxicity of flavonoids from the flowers of Chromolaena odorata. Arch Pharm Res 27(5): 507-511.
TALEB-CONTINI SH, KANASHIRO A, KABEYA LM, POLIZELLO ACM, LUCISANO-VALIM YM, OLIVEIRA DCR. 2006. Immunomodulatory effects of methoxylated flavonoids from two Chromolaena species: structure-activity relationships. Phytother Res 20(7): 573-575.
TALEB-CONTINI SH, SALVADOR MJ, BALANCO JMF, ALBUQUERQUE S, OLIVEIRA DCR. 2003. Short communication: Antiprotozoal effect of crude extracts and flavonoids isolated from Chromolaena hirsuta (Asteraceae). Inflammopharmacology 13(5- 6): 479-484.
TIWARI U, RASTOGI B, SINGH P, SARAF DK, VYAS SP. 2004. Immunomodulatory effects of aqueous extract of Tridax precumbens in experimental animals. J Ethnopharm 92: 113-119.
VITAL PG. 2008. Antimicrobial activity and cytotoxicity of Chromolaena odorata (Lf.) King & Robinson and Uncaria perrottetii (A. Rich) Merr. extracts. [MS Thesis] Quezon City, Philippines: University of the Philippines, Diliman. (Available at the UPD Library)
VITAL PG, RIVERA WL. 2009. Antimicrobial activity and cytotoxicity of Chromolaena odorata (Lf.) King & Robinson and Uncaria perrottetii (A. Rich) Merr. extracts. J Med Plant Res 3(7): 511-518.
ZELIKOFF JT. 1997. Techniques in fish immunology (Workshop Manual). Australia: University of Tasmania.
ZELIKOFF JT, WANG W, ISLAM N, TWERDOK LE, CURRY M, BEAMAN J, FLESHER E. 1996. Assays of reactive oxygen intermediates and antioxidant enzymes in medaka (Oryza latipes): Potential biomarkers for predicting the effects of environmental pollution. Techniques in Aquatic Toxicology. (G. Ostrander ed.) Florida, USA: CRC Press. p. 178-206.
ZHU X-L, CHEN A-F, LIN B-Z. 2007. Ganoderma lucidum polysaccharides enhance the function of immunological effector cells in immunosuppressed mice. J Ethnopharm 111: 219-226.