Chemical characterization and cytotoxic, genotoxic, and mutagenic properties of Baccharis trinervis (Lam, Persoon) from Colombia and Brazil.

PHARMACOLOGY RELEVANCE: Baccharis trinervis (Lam, Persoon) leaves are used in the traditional medicine for the treatment of high fevers, edema, inflammation, sores and muscle cramps, snakebites and as antiseptic. AIM OF THE STUDY: To investigate the cytotoxic, genotoxic, and mutagenic effects of extracts and fractions of B. trinervis from Brazil and Colombia in Chinese Hamster Ovary (CHO) cells, and to examine the mutagenic activity in Salmonella typhimurium. MATERIAL AND METHODS: Aqueous extracts (AE) of aerial parts of B. trinervis from Brazil (B) and Colombia (C) were fractioned in ethyl acetate fraction (EAF), butanol extract (BF), and aqueous residue fraction (ARF). Qualitative chemical screening and determination of total flavonoid content were made. Identification of chemical constituents was performed by High Performance Liquid Chromatography (HPLC) and High Resolution Mass Spectrometry (HRMS). For the in vitro tests, CHO cells were treated for 3h with extracts and fractions. The cytotoxic activity was evaluated by clonal survival and 3-(4.5-dimethylthiazole-2-yl)-2.5-biphenyl tetrazolium bromide reduction assay (MTT). Genotoxic and mutagenic effects were evaluated by the alkaline comet assay and Cytokinesis-blockage micronucleus test (CBMN), respectively. Additionally, Salmonella/microsome assay was carried out to determinate the mutagenic effects in EAF from Brazil and Colombia. RESULTS: Phytochemical analyses indicated the presence of saponins and flavonoids. AE and EAF were the samples with the highest quantity of total flavonoids. HPLC showed the presence of luteolin only in AEC, and caffeic acid, ellagic acid, rosmarinic acid, and rutin were identified in AEB and AEC (AEC>AEB). The HRMS in positive mode of EAFB and EAFC showed presence of two carboxylic acids, coumarin, and two terpenoids. In addition, were identified one terpenoid and two carboxylic acids in AE, BF and ARF of B. trinervis from both countries in negative mode. Dose-dependent cytotoxic effects were observed in CHO cells treated with B. trinervis extracts and fractions by using clonal survival and MTT at concentrations higher than 0.05mg/mL. All the extracts and fractions induced DNA strand breaks in CHO cells with dose-dependent response, mostly EAFB and EAFC. The EAF from Brazil and Colombia showed mutagenic effect at 0.5mg/mL, while the other fractions did not show a significant difference in relation to the control. No mutagenic effects were found in EAF from both countries by the Salmonella/microsome assay. CONCLUSIONS: Cytotoxic and genotoxic effects were demonstrated in all extracts and fractions used, although only EAF showed mutagenic effects by CBMN, but not by Salmonella/microsome assay. Our results suggest that flavonoids, phenylpropanoids, coumarins, and diterpenes may be responsible for the cytotoxic, genotoxic and mutagenic effects observed.

Chemical and toxicological effects of medicinal Baccharis trimera extract from coal burning area.

The entire process of power generation, extraction, processing and use of coal strongly impact water resources, soil, air quality and biota leads to changes in the fauna and flora. Pollutants generated by coal burning have been contaminating plants that grow in area impacted by airborne pollution with high metal contents. Baccharis trimera is popularly consumed as tea, and is widely developed in Candiota (Brazil), one of the most important coal burning regions of the Brazil. This study aims to investigate the phytochemical profile, in vivo genotoxic and mutagenic potential of extracts of B. trimera collected from an exposed region to pollutants generated by coal burning (Candiota City) and other unexposed region (Bagé City), using the Comet assay and micronucleus test in mice and the Salmonella/microsome short-term assay. The HPLC analyses indicated higher levels of flavonoids and phenolic acids for B. trimera aqueous extract from Bagé and absence of polycyclic aromatic hydrocarbons for both extracts. The presence of toxic elements such as cobalt, nickel and manganese was statistically superior in the extract from Candiota. For the Comet assay and micronucleus test, the mice were treated with Candiota and Bagé B. trimera aqueous extracts (500-2000 mg/kg). Significant genotoxicity was observed at higher doses treated with B. trimera aqueous extract from Candiota in liver and peripheral blood cells. Micronuclei were not observed but the results of the Salmonella/microsome short-term assay showed a significant increase in TA98 revertants for B. trimera aqueous extract from Candiota. The extract of B. trimera from Candiota bioacumulated higher levels of trace elements which were associated with the genotoxic effects detected in liver and peripheral blood cells.

Neuropharmacological and genotoxic evaluation of ethanol extract from Erythrina falcata leaves, a plant used in Brazilian folk medicine

The aim of the present study was to determine neurobehavioral and genotoxic activities of ethanol extract of Erythrina falcata Benth., Fabaceae, leaves on rats. Animals were treated with ethanol extract of E. falcata (100, 300 or 500 mg/kg; i.p.) and the open field and elevated plus-maze tasks were used as behavioral models to investigate a possible effect on the locomotor and exploratory activity and anxiety, respectively. Genotoxic effect was investigated using the Comet assay. Ethanol extract of E. falcata leaves decreased the number of crossings and rearings in the open field task and increased the latency to start locomotion, though it was not able to affect habituation to apparatus measured 24h after the first session. Behavioral parameters in the plus-maze test were not affected by E. falcata. Ethanol extract did not increase damage index and damage frequency in blood or brain, indicating no genotoxic effect. The results suggest that ethanol extract of E. falcata leaves was able to affect locomotion, exploration, and motivation of animals without anxiolytic/anxiogenic effect, indicating a possible depressant action on the central nervous system. Furthermore, the lack of DNA damage in brain is an indicative that ethanol extract of E. falcata leaves may not induce neurotoxic effects.

The antigenotoxic activity of latex from Himatanthus articulatus

Himatanthus articulatus (Vahl) Woodson (Apocynaceae) is a native plant to the Amazon popularly used to treat ulcers, tumors, inflammations, cancer, syphilis and malaria. The aim of the present study was to investigate the in vivo genotoxic/antigenotoxic and mutagenic potential of this plant, using the comet and the micronucleus assays in mice. Female and male adult mice were treated with different doses of H. articulatus latex by gavage for two consecutive days. For the experiments, the latex was serially diluted with water to 1:2 (D1); 1:4 (D½) and 1:8 (D») and administered to the animals. The blood slides were exposed to hydrogen peroxide (ex vivo) to evaluate antigenotoxic effect. Under the experimental conditions used in this study, the latex of H. articulatus did not increase the frequency of DNA damage as measured by the comet assay and micronucleus test in treated mice, indicating a non-genotoxic and non-mutagenic activity. In relation to the antigenotoxicity, latex exerted protective effect against DNA damage induced by hydrogen peroxide. Therefore, our results add new information about the antigenotoxic potential of H. articulatus latex, which is popularly used in the Amazon to treat different pathologies.

Evaluation of effects of dichloromethane fraction from Platonia insignis on pilocarpine-induced seizures

The objective of present study was to evaluate the antioxidant and anticonvulsant activities of dichloromethane fraction (DMF) from Platonia insignis Mart., Clusiaceae. The DMF from P. insignis (2 mg/kg) was tested by intraperitoneal (i.p.) to evaluate effects on lipid peroxidation level, nitrite formation, as well as on locomotor and anticonvulsant activities. Wistar rats were treated with, (saline/Tween 80 0.5 percent, i.p., control group), DMF (2 mg/kg, i.p., DMF group), pilocarpine (400 mg/kg, i.p., P400 group), or the combination of DMF (2 mg/kg, i.p.) and pilocarpine (400 mg/kg, i.p., DMF plus P400). After the treatments all groups were observed for 24 h. In P400 group rats there was a decrease in the motor activity when compared with control group. In DMF plus P400 co-administered rats was observed an increase in motor activity when compared with P400 group. In P400 group rats there was a significant increase in lipid peroxidation and nitrite levels. In DMF plus P400 co-administered rats, antioxidant treatment significantly reduced the lipid peroxidation level and nitrite content after seizures. Previous findings strongly support the hypothesis that oxidative stress occurs in rat striatum during pilocarpine-induced seizures, and our results imply that strong neuprotective effect on this brain region could be achieved using DMF from P. insignis.

Antiproliferative effect of a traditional remedy, Himatanthus articulatus bark, on human cancer cell lines.

ETHNOPHARMACOLOGICAL RELEVANCE: Himatanthus articulatus (Vahl) Woodson (Apocynaceae) is a tree occurring in the Amazon region. The local population uses its bark against to external tumors and cancer. AIM OF THE STUDY: Evaluated the antiproliferative activity of the crude extract and their fractions against human tumors cells. MATERIALS AND METHODS: The antiproliferative responses of the crude extract and their fractions were colorimetrically evaluated by sulforhodamine B (SRB) assay against HT-29 (colon adenocarcinoma); NCI-H460 (non-small cell lung carcinoma); MCF-7 (breast cancer); OVCAR-3 (ovarian adenocarcinoma) and RXF-393 (renal cell carcinoma) as well as against NIH-3T3 (mouse embryo fibroblast cell). RESULTS: The cytotoxicity data from the crude extract allowed considering active only in the NCI-H460 cell line. However, the antiproliferative activity was much more pronounced at the chloroformic fraction in all cell lines tested. The butanolic fraction demonstrated activity against to HT-29, MCF-7, RXF-393 and OVCAR-3 cells. The ethyl acetate fraction demonstrated activity only in RXF-393 and the aqueous residue did not present antiproliferative effect. CONCLUSIONS: Through the popular use we find that the crude extracts of Himatanthus articulatus bark displayed weak cytotoxic. However, the butanolic fraction showed to be active only against to tumor cell lines while chloroformic fraction possesses high activity being similar to the positive control. Both fractions have been selected for future bio-guided fractionation and isolation of active compounds.