title: Bioassay-guided evaluation of Dioscorea villosa - an acute and subchronic toxicity, antinociceptive and anti-inflammatory approach.
abstract: BACKGROUND
  Dioscorea villosa (DV) has been used in Brazil as an alternative medicine to attenuate menopause symptoms, as well as for the treatment of joint pain and rheumatoid arthritis. In spite of the popular use of DV for the treatment of various disorders, there are limited scientific data regarding safety aspects of this herb. In this regard, we carried out to evaluated both antinociceptive and anti-inflammatory activities in experimental models and assess the toxic effects of the acute (single dose) and subchronic (30 days) oral administration of dry extract of Dioscorea villosa in rodents.
  
  
  METHODS
  The LC analyses were performed to assess the presence of the diosgenin in samples of DV. The antinociceptive study of DV was performed using models of acetic acid-induced writhing and formalin-induced pain in mice. The anti-inflammatory study was accomplished by leukocyte migration to the peritoneal cavity. A dry extract of DV was tested at doses of 100, 200 and 400 mg/kg (per os or p.o.). The toxicological properties of the dry extract were evaluated by toxicity assays of acute (5 g/kg, single dose) and subchronic (1 g/kg/day, 30 days) treatment. Haematological, biochemical, and histopathological parameters were studied. The results are expressed as mean ± S.D., and statistical analysis of the data were performed with the Student's t-test or one-way analysis of variance (ANOVA) followed by Tukey's test. In all cases differences were considered significant if p < 0.05.
  
  
  RESULTS
  HPLC-DAD analysis of the extract from DV revealed the presence of diosgenin as the major compound. Doses of 200 and 400 mg⁄kg significantly reduced the amount of acetic acid-induced writhing in relation to the vehicle (p < 0.0001). In the first phase, using the formalin-induced neurogenic pain test, only the 400 mg/kg dose of DV showed significant inhibition of neurogenic pain (p < 0.001). In the second phase, 200 and 400 mg/kg of DV showed significant inhibition of inflammatory pain (p < 0.0001). Significant inhibition of leukocyte migration was observed with doses of 100 (p < 0.001), 200 (p < 0.01) and 400 mg/kg (p < 0.01). Haematological, biochemical and histopathological data obtained in both acute and subchronic toxicological assays revealed only unremarkable changes, which are unlikely to indicate DV toxicity with oral administration.
  
  
  CONCLUSIONS
  We found that DV possesses antinociceptive and anti-inflammatory properties in rodent models. In addition, no acute or subchronic toxicity was evident when the herbal extract was administered orally. These results supporting the folkloric usage of the plant to treat various inflammatory diseases.
authors.forename: Claudio Moreira
authors.initials: CM
authors.lastname: Lima
authors.forename: Adriana Karla
authors.initials: AK
authors.lastname: Lima
authors.forename: Marcelia G Dória
authors.initials: MG
authors.lastname: Melo
authors.forename: Mairim Russo
authors.initials: MR
authors.lastname: Serafini
authors.forename: Dênisson Lima
authors.initials: DL
authors.lastname: Oliveira
authors.forename: Enrik Barbosa
authors.initials: EB
authors.lastname: de Almeida
authors.forename: Rosana Souza Siqueira
authors.initials: RS
authors.lastname: Barreto
authors.forename: Paulo Cesar de Lima
authors.initials: PC
authors.lastname: Nogueira
authors.forename: Valéria Regina de Souza
authors.initials: VR
authors.lastname: Moraes
authors.forename: Edica Ramone Andrade
authors.initials: ER
authors.lastname: Oliveira
authors.forename: Ricardo Luiz Cavalcanti
authors.initials: RL
authors.lastname: de Albuquerque
authors.forename: Lucindo J
authors.initials: LJ
authors.lastname: Quintans-Júnior
authors.forename: Adriano Antunes Souza
authors.initials: AA
authors.lastname: Araújo
mesh_terms: D019342:Acetic Acid; D000284:Administration, Oral; D000700:Analgesics / Q000009:adverse effects / Q000494:pharmacology / Q000627:therapeutic use*; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000009:adverse effects / Q000494:pharmacology / Q000627:therapeutic use*; D001681:Biological Assay; D027742:Dioscorea / Q000009:adverse effects / Q000737:chemistry*; D004144:Diosgenin / Q000009:adverse effects / Q000032:analysis / Q000494:pharmacology / Q000627:therapeutic use*; D005260:Female; D005557:Formaldehyde; D007249:Inflammation / Q000188:drug therapy*; D007962:Leukocytes / Q000378:metabolism; D008297:Male; D051379:Mice; D010146:Pain / Q000139:chemically induced / Q000188:drug therapy*; D008517:Phytotherapy; D010936:Plant Extracts / Q000009:adverse effects / Q000494:pharmacology / Q000627:therapeutic use; D051381:Rats; D017208:Rats, Wistar
chemical_list: D000700:Analgesics; D000893:Anti-Inflammatory Agents; D010936:Plant Extracts; D005557:Formaldehyde; D004144:Diosgenin; D019342:Acetic Acid
references.citation: J Pain. 2007 Feb;8(2):127-36
references.pmid: 16949880
references.citation: Eur J Pharmacol. 1986 Jan 14;120(1):33-41
references.pmid: 3948914
references.citation: Menopause. 2008 Sep-Oct;15(5):819-20
references.pmid: 18779673
references.citation: Eur J Pharmacol. 1980 Jan 11;61(1):17-24
references.pmid: 7353582
references.citation: J Med Food. 2011 Nov;14(11):1389-96
references.pmid: 21663477
references.citation: Pain. 1987 Jul;30(1):103-114
references.pmid: 3614974
references.citation: Fed Proc. 1987 Jan;46(1):118-26
references.pmid: 3100339
references.citation: Eur J Pharmacol. 2007 Dec 8;576(1-3):171-9
references.pmid: 17719028
references.citation: Basic Clin Pharmacol Toxicol. 2010 Dec;107(6):949-57
references.pmid: 20849525
references.citation: Pain. 1992 Oct;51(1):5-17
references.pmid: 1454405
references.citation: J Ethnopharmacol. 2001 May;75(2-3):141-64
references.pmid: 11297844
references.citation: Diabetes. 2000 Sep;49(9):1543-51
references.pmid: 10969839
references.citation: Clin Chim Acta. 2001 Oct;312(1-2):213-9
references.pmid: 11580929
references.citation: Pharmacol Rev. 2001 Dec;53(4):597-652
references.pmid: 11734620
references.citation: Biol Pharm Bull. 2008 Oct;31(10):1977-80
references.pmid: 18827367
references.citation: BMC Complement Altern Med. 2012 Aug 06;12:117
references.pmid: 22866906
references.citation: Eur J Immunol. 1995 Apr;25(4):947-51
references.pmid: 7737298
references.citation: Braz J Med Biol Res. 1988;21(2):341-3
references.pmid: 3203167
references.citation: Br J Pharmacol. 1973 May;48(1):88-96
references.pmid: 4146764
references.citation: Menopause. 2008 Mar-Apr;15(2):374-81
references.pmid: 17882009
references.citation: Toxicology. 2002 Oct 15;179(3):183-96
references.pmid: 12270592
references.citation: Psychopharmacology (Berl). 1994 Jul;115(3):311-9
references.pmid: 7871070
references.citation: Vascul Pharmacol. 2010 Nov-Dec;53(5-6):273-80
references.pmid: 20932938
references.citation: Pharm Biol. 2010 Apr;48(4):411-6
references.pmid: 20645719
references.citation: Biol Pharm Bull. 2008 May;31(5):1017-20
references.pmid: 18451538
references.citation: Phytomedicine. 2001 Jul;8(4):310-22
references.pmid: 11515723
references.citation: Proc Soc Exp Biol Med. 1962 Dec;111:544-7
references.pmid: 14001233
references.citation: Menopause. 2002 Sep-Oct;9(5):329-34
references.pmid: 12218721
references.citation: BMC Complement Altern Med. 2012 Aug 11;12:120
references.pmid: 22883637
references.citation: J Ethnopharmacol. 2012 Jan 31;139(2):330-6
references.pmid: 22138517
references.citation: Nat Immunol. 2005 Dec;6(12):1182-90
references.pmid: 16369557
references.citation: Food Chem Toxicol. 2008 Sep;46(9):3122-31
references.pmid: 18662738
references.citation: Int J Toxicol. 2004;23 Suppl 2:49-54
references.pmid: 15513824
references.citation: J Ethnopharmacol. 2006 Oct 11;107(3):370-3
references.pmid: 16697541
references.citation: Br J Pharmacol. 1987 Aug;91(4):823-32
references.pmid: 2822190
references.citation: Climacteric. 2001 Jun;4(2):144-50
references.pmid: 11428178
references.citation: J Appl Physiol (1985). 2003 Jun;94(6):2127-34
references.pmid: 12547845
references.citation: Am J Physiol. 1997 Aug;273(2 Pt 1):G355-64
references.pmid: 9277414
references.citation: Menopause. 2009 Jan-Feb;16(1):156-63
references.pmid: 18791483