Assessment of the in vitro and in vivo genotoxic and antigenotoxic effects of pimaradienoic acid in mammalian cells.

The pimarane-type diterpene, pimaradienoic acid (PA), is known for its diverse biological properties such as antimicrobial, anti-inflammatory and trypanocidal. A preliminary study was undertaken to investigate in vitro the free radical-scavenging potential of PA. In addition, the genotoxic potential of PA and its ability to modulate genotoxicity induced by doxorubicin (DXR) and methyl methanesulfonate (MMS) were studied in Chinese hamster lung fibroblasts (V79 cells) and in male Swiss mice using the comet and micronucleus assays. The DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) assay showed that PA exerted no antioxidant activity when compared to quercetin. The colony-forming assay using V79 cells showed that PA was cytotoxic at concentrations >5.0µg/mL. Therefore, concentrations of 0.625, 1.25, 2.5, and 5.0µg/mL were used for evaluation of the genotoxic and antigenotoxic potential of PA in V79 cells. For genotoxic and antigenotoxic assessment in Swiss mice, three PA doses were tested (20, 40, and 80mg/kg body weight) based on the solubility limit of the diterpene in dimethylsulfoxide and water. The in vitro results demonstrated that PA induced DNA damage at concentrations of 2.5 and 5.0µg/mL in the comet assay. However, no genotoxic effect was observed in the micronucleus test using V79 cells. In the in vivo evaluation of genotoxicity, a significant increase in the frequency of DNA damage was observed in hepatocytes of animals treated with the highest PA dose (80mg/kg) when compared to the control group, but this difference was not seen in the micronucleus test. Furthermore, PA significantly reduced the frequency of DXR- and MMS-induced micronuclei and extent of DNA damage in in vitro and in vivo test systems.

Effects of propolis crude hydroalcoholic extract on chromosomal aberrations induced by doxorubicin in rats.

Propolis has been reported to display a broad spectrum of biological activities such as anticancer, antioxidant, anti-inflammatory, antibiotic and antifungal properties, among others. There is great interest not only in the determination of the chemical composition of propolis but also in the understanding of the mechanisms related to its therapeutic actions. In this respect, the aim of the present investigation was to study the influence of both simultaneous (6, 12 and 24 mg/kg b. w.) and subacute (12 mg/kg b. w.) treatment with a crude hydroalcoholic extract of propolis on the frequency of chromosome aberrations induced by the chemotherapeutic agent doxorubicin (DXR) in Wistar rat bone marrow cells. HPLC analysis of the crude extract allowed the quantification of the phenolic compounds: caffeic acid, P-coumaric acid, aromadendrin 4'-methyl ether, 3-prenyl- P-coumaric acid (drupanin), isosakuranetin, kaempferide, 3,5-diprenyl- P-coumaric acid (artepellin C), baccharin and 2,2-dimethyl-6-carboxyethenyl-2 H-1-benzopyran. A total of 100 metaphase cells/animal were analyzed for chromosome aberration frequency and 1000 cells/animal were counted to obtain the mitotic index. The results showed that the dose of 12 mg propolis/kg b. w., administered either as a single dose or as subacute treatment, caused a statistically significant decrease in the frequency of chromosome damage induced by DXR compared to the group treated only with DXR. This reduction might be, in part, due to the presence of phenolic compounds in the studied propolis, which are able to capture free radicals produced by chemotherapeutic agents such as DXR.