Clastogenicity of Piper cubeba (Piperaceae) seed extract in an in vivo mammalian cell system

Abstract

The plant Piper cubeba is widely distributed in tropical and subtropical regions and is used medically for various purposes but has not yet been evaluated for genotoxicity. We used male and female Swiss mice and Wistar rats and the comet assay and micronucleus test to investigate the mutagenic potential of a crude extract of P. cubeba seeds. The rodents were administered 0.5 g kg -1 , 1.0 g kg -1 and 1.5 g kg -1 of the extract by gavage. For the Swiss mice, peripheral blood was collected 24 h after treatment for the comet assay, and at 48 and 72 h for the micronucleus test. For the Wistar rats, peripheral blood and hepatic cells were collected for the comet assay and bone marrow cells were collected for the micronucleus test 24 h after treatment. At 1.5 g kg -1 , the highest dose tested, the extract induced a statistically significant increase in both the mean number of micronucleated polychromatic erythrocytes and the level of DNA damage in the rodent cell types analyzed. Under our experimental conditions, the P. cubeba seed extract was genotoxic in vivo when administered orally to mice and rats.

Genotoxic effects of (−)‐cubebin in somatic cells of mice

“…On the other hand, crude P . cubeba seed extract was genotoxic in rodents (Junqueira et al ., 2007). Data about the possible mutagenicity or genotoxicity effects from the main components of the above extract have not been reported before.…”

“…Regarding genotoxicity, Junqueira et al . (2007) reported that the crude P . cubeba seed extract was genotoxic in vivo when administered orally to mice and rats.…”

“…Choi and Hwang (2003) demonstrated significant anti-inflammatory and analgesic activities of the methanolic extract from fruits of P. cubeba. Regarding genotoxicity, Junqueira et al (2007) reported that the crude P. cubeba seed extract was genotoxic in vivo when administered orally to mice and rats.…”

Determination of the Mutagenicity Potential of Dillsun Herbal Medicine by Single Cell Gel Electrophoresis in Rat Hepatocytes

“…The parameters used for statistical analysis were the tail moment and tail length (distance from the middle of nucleoid core to the end of the tail). Cells were visually scored into comet classes according to their tail characteristics: Class 0 = there was no tail, Class 1 = the tail was shorter than the diameter of the head (nucleus), Class 2 = the tail was 1 to 2x the diameter of the nucleus and Class 3 = the tail was longer than 2x the diameter of the nucleus (15)(16)(17). Lack of nucleus in DNA or very wide tailed DNA was deleted from this study because it could be a cell (18,19).…”

Determination of the Mutagenicity Potential of Sankol Herbal Medicine by Single Cell Gel Electrophoresis in Rat Hepatocytes in Comparison With H2O2

“…The extent and distribution of DNA damage indicated by comet assay was evaluated by examining cells. The cells were visually scored into comet classes according to tail size class (16, 17, 18) Class 0 = no tail, Class 1 = tail shorter than the diameter of the head (nucleous) , Class 2 = tail length 1 to 2x the diameter of the head and Class 3 = tail longer than 2x the diameter of the head. Comet without head and those with nearly all the DNA in the tail or with a very wide tail, were excluded from the evaluation because they probably represented dead cells (19, 20) tail length and the mutagenic Index were calculated based on the following formula MI = (0NMC+ 1SMC+ 2MMC+ 3LMC) /200, or could be expressed as NMC = No migration cells (score 0), SMC = Short migration cells (score 1) ,MMC = Medium migration cells (score 2), LMC = Long migration cells (score 3).…”