Planta Med 2010; 76(9): 935-940
DOI: 10.1055/s-0029-1240813
Biological Screening
Original Papers
© Georg Thieme Verlag KG Stuttgart · New York

Anti-Angiogenic and Cytotoxicity Studies of Some Medicinal Plants

Kwok-Wen Ng1 , Salizawati Muhamad Salhimi1 , Amin Malik Shah Abdul Majid1 , Kit-Lam Chan1
  • 1School of Pharmaceutical Sciences, Universiti Sains Malaysia, 11800 Penang, Malaysia
Further Information

Publication History

received Sept. 11, 2009 revised Dec. 11, 2009

accepted Dec. 24, 2009

Publication Date:
28 January 2010 (online)

Abstract

Angiogenesis plays an important role in tumor formation and proliferation. The development of anti-angiogenic agents to block new blood vessel growth will inhibit metastasis and induce apoptosis of the cancer cells. Nine medicinal plants, Strobilanthes crispus, Phyllanthus niruri, Phyllanthus pulcher, Phyllanthus urinaria, Ailanthus malabarica, Irvingia malayana, Smilax myosotiflora, Tinospora crispa and Blumea balsamifera were screened for anti-angiogenic properties using the rat aortic ring assay. Of these, the methanol extracts of Phyllanthus species and Irvingia malayana exhibited the highest activity. At 100 µg/mL, P. pulcher, P. niruri, P. urinaria and I. malayana recorded an inhibition of 78.8 %, 59.5 %, 56.7 % and 46.4 %, respectively, against rat aortic vascular growth. Their activities were further investigated by the tube formation assay involving human umbilical vein endothelial cells (HUVEC) on Matrigel. I. malayana, P. niruri and P. urinaria showed a significant decrease of 45.5, 37.9 and 35.6 %, respectively, whilst P. pulcher showed a much lower decrease of 15.5 % when compared with that of the rat aortic ring assay. All the plant extracts were evaluated for cytotoxicity on a panel of human cancer cell lines using the MTT assay. None of them displayed acute cytotoxicity. The HPLC of P. niruri, P. urinaria and P. pulcher indicated the extracts contained some identical chromatographic peaks of lignans. Further fractionation of I. malayana yielded betulinic acid reported in this plant for the first time and at 100 µg/mL it exhibited a 67.3 % inhibition of vessel outgrowth and 46.5 % inhibition of tube formation.

References

  • 1 Tosetti F, Ferrari N, Flora S D, Albini A. “Angioprevention”: angiogenesis is a common and key target for cancer chemopreventive agents.  FASEB J. 2002;  16 2-14
  • 2 Folkman J. Anti-angiogenesis: new concept for therapy of solid tumors.  Ann Surg. 1972;  175 409-416
  • 3 Mariotti M, Maier J A M. Angiogenesis: an overview. Forough R New frontiers in angiogenesis. Berlin; Springer 2006: 1-29
  • 4 Abu-Bakar M F, Teh A H, Rahmat A, Othman F, Hashim N, Fakurazi S. Antiproliferative properties and antioxidant activity of various types of Strobilanthes crispus tea.  Int J Cancer Res. 2006;  2 152-158
  • 5 Ismail M, Manickam E, Danial A M, Rahmat A, Yahaya A. Chemical composition and antioxidant activity of Strobilanthes crispus leaf extract.  J Nutr Biochem. 2000;  11 536-542
  • 6 Bagalkotkar G, Sagineedu S R, Saad M S, Stanslas J. Phytochemicals from Phyllanthus niruri Linn. and their pharmacological properties: a review.  J Pharm Pharmacol. 2006;  58 1559-1570
  • 7 Ajaiyeoba E, Kingston D. Cytotoxicity evaluation and isolation of a chroman derivative from Phyllanthus amarus aerial part extract.  Pharm Biol. 2006;  44 668-671
  • 8 Joy K L, Kuttan R. Inhibition by Phyllanthus amarus of hepatocarcinogenesis induced by N-nitrosodiethylamine.  J Clin Biochem Nutr. 1998;  24 133-139
  • 9 Rajeshkumar N V, Kuttan R. Phyllanthus amarus extract administration increases the life span of rats with hepatocellular carcinoma.  J Ethnopharmacol. 2000;  73 215-219
  • 10 Calixto J B, Santos A R S, Paulino N, Cechinel Filho V, Yunes R A. The plants of the genus Phyllanthus as a potential source of new drugs.  Cienc Cult. 1997;  9 422-432
  • 11 Kumar D, Kumar S, Vasudeva N. A review: chemistry and biological activities of genus Phyllanthus.  Int J Plant Sci. 2006;  1 339-346
  • 12 Huang S T, Yang R C, Lee P N, Yang S H, Liao S K, Chen T Y, Pang J H S. Anti-tumor and anti-angiogenic effects of Phyllanthus urinaria in mice bearing Lewis lung carcinoma.  Int Immunopharmacol. 2006;  6 870-879
  • 13 Bharali R, Tabassum J, Azad M R H. Chemopreventive action of Phyllanthus urinaria Linn on DMBA-induced skin carcinogenesis in mice.  Indian J Exp Biol. 2003;  41 1325-1328
  • 14 Giridharan P, Somasundaram S T, Perumal K, Vishwakarma R A, Karthikeyan N P, Velmurugan R, Balakrishnan A. Novel substituted methylenedioxy lignan suppresses proliferation of cancer cells by inhibiting telomerase and activation of c-myc and caspases leading to apoptosis.  Br J Cancer. 2002;  87 98-105
  • 15 Sureban S M, Subramaniam D, Rajendran P, Ramanujam R P, Dieckgraefe B K, Houchen C W, Anant S. Therapeutic effects of Phyllanthus species: induction of TNF-alpha-mediated apoptosis in HepG2 hepatocellular carcinoma cells.  Am J Pharmacol Toxicol. 2006;  1 65-71
  • 16 Nguyen-Pouplin J, Tran H, Phan-Tuyet A, Dolecek C, Farrar J, Tran-Tinh H, Caron P, Bodo B, Grellier P. Antimalarial and cytotoxic activities of ethnopharmacologically selected medicinal plants from South Vietnam.  J Ethnopharmacol. 2007;  109 417-427
  • 17 Nessa F, Ismail Z, Mohamed N, Haris M R H M. Free radical-scavenging activity of organic extracts and of pure flavonoids of Blumea balsamifera DC leaves.  Food Chem. 2004;  88 243-252
  • 18 Osaki N, Koyano T, Kowithayakorn T, Hayashi M, Komiyama K, Ishibashi M. Sesquiterpenoids and plasmin-inhibitory flavonoids from Blumea balsamifera.  J Nat Prod. 2005;  68 447-449
  • 19 Noor-Rain A, Khozirah S, Mohd-Ridzuan M A R, Ong B K, Rohaya C, Rosilawati M, Hamdino I, Badrul A, Zakiah I. Antiplasmodial properties of some Malaysian medicinal plants.  Trop Biomed. 2007;  24 29-35
  • 20 Cichewicz R H, Kouzi S A. Chemistry, biological activity, and chemotherapeutic potential of betulinic acid for theprevention and treatment of cancer and HIV infection.  Med Res Rev. 2004;  24 90-114
  • 21 Berger A C, Wang X Q, Zalatoris A, Cenna J, Watson J C A. Murine model of ex vivo angiogenesis using aortic disks grown in fibrin clot.  Microvasc Res. 2004;  68 179-187
  • 22 Mosmann T. Rapid colorimetric assay for cellular growth and survival application to proliferation and cytotoxicity assays.  J Immunol Methods. 1983;  65 55-63
  • 23 Hang C C, Lien Y C, Liu K, Chen C S, Lee S S. Lignans from Phyllanthus urinaria.  Phytochemistry. 2003;  63 825-833
  • 24 Murugaiyah V, Chan K L. Determination of four lignans in Phyllanthus niruri L. by a simple high-performance liquid chromatography method with florescence detection.  J Chromatogr A. 2007;  1154 198-204
  • 25 Anupam S, Rayneet T S, Sukhdev S H. Determination of phyllanthin and hypophyllanthin by high-performance liquid chromatography in Phyllanthus amarus.  Phytochem Anal. 1993;  4 226-229
  • 26 Murugaiyah V, Chan K L. Antihyperuricemic lignans from the leaves of Phyllanthus niruri.  Planta Med. 2006;  72 1-6
  • 27 Kwon H J, Shim J H, Kim J H, Cho H Y, Yum Y N, Kim S H, Yu J. Betulinic acid inhibits growth factor-induced in-vitro angiogenesis via the modulation of mitochondrial function in endothelial cell.  Jpn J Cancer Res. 2002;  93 417-425
  • 28 Grants D S, Kinsella J L, Kibbey M C, LaFlamme S, Burbelo P D, Goldstein A L, Kleinman H K. Matrigel induces thymosin beta4 gene in differentiating endothelial cell.  J Cell Sci. 1995;  108 3685-3694
  • 29 Villaschi S, Nicosia R F. Angiogenesis role of endogenous basic fibroblast growth factors released by rat aorta after injury.  Am J Pathol. 1993;  143 181-190
  • 30 Pisha E, Chai H, Lee L S, Chagwedera T E, Farnworth N R, Cordell G A, Beecher C W, Fong H H, Kinghorn A D, Brown D M. Discovery of betulinic acid as a selective inhibitor of human melanoma that functions by induction of apoptosis.  Nat Med. 1995;  1 1046-1051
  • 31 Zuco V, Supino R, Righetti R C, Cleris L, Marchesi E, Gambacorti-Passerini C, Formelli F. Selective cytotoxicity of betulinic acid on tumor cell lines, but not on normal cells.  Cancer Lett. 2002;  175 17-25

Prof. Dr. Kit-Lam Chan

School of Pharmaceutical Sciences
Universiti Sains Malaysia

11800 Penang

Malaysia

Phone: + 60 46 53 38 88 ext. 26 96

Email: kitlamc@gmail.com

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