Elsevier

Journal of Ethnopharmacology

Volume 230, 10 February 2019, Pages 109-116
Journal of Ethnopharmacology

Safety assessment of Piper cernuum Vell. (Piperaceae) leaves extract: Acute, sub-acute toxicity and genotoxicity studies

https://doi.org/10.1016/j.jep.2018.10.035Get rights and content

Abstract

Ethnopharmacological relevance

Piper cernuum Vell (Piperaceae) is a native species from Atlantic rain forest, popularly known as pariparoba. Its leaves have been commonly used by rural and urban communities from State of São Paulo, Brazil, to treat pain (orally and topically), and hepatic and renal complications.

Aim of the study

In this study we evaluated the acute and sub-acute toxicity, genotoxicity and mutagenicity of hydroalcoholic extract obtained from P. cernuum leaf using in vivo and in vitro methods.

Material and methods

In the acute toxicity study, mice were orally treated with P. cernuum extract (2000 mg/kg, p.o.). General behavior and mortality were observed for up to 14 days. In the sub-acute toxicity study, P. cernuum extract was given orally as a single administration to the rats at doses of 50 or 250 mg/kg/day, for 28 days. General behavior, body weight, biochemical and hematological parameters, organ coefficients and pathological morphology were analyzed. The P. cernuum mutagenicity was evaluated using mammalian cell micronucleus assay. Additionally, in vitro toxicity profile of the extract was assessed through cytotoxicity, hemolytic activity, and genotoxicity assay.

Results

Data from comet assay demonstrates that high concentrations of P. cernuum extract induce genotoxicity. However, no evidence of hemolytic, cytotoxic or mutagenicity activity was found. In addition, the acute and sub-acute toxicity studies did not show significant changes in body weight, general behavior, hematology and biochemical parameters, organ weight and liver and kidney histopathological analysis.

Conclusions

Together, the results herein obtained indicate that P. cernuum leaves extract did not present significant toxicity when administered to male or female rats. Additionally, no significant alteration in hematological, biochemical and morphological parameters were observed. Data obtained in vitro shows that extract did not present cytotoxicity and mutagenicity. However, the extract induces in vitro genotoxicity, but in high concentration. Further studies are necessary to evaluate the safety of long-term exposure to P. cernuum leaves extract added to in vivo genotoxicity.

Introduction

The use of natural products has considerably increased in recent years, and this increase is associated with the fact that the population believes that natural products did not present toxic effects (Moreira et al., 2010, Yuan et al., 2016). However, most of natural products has not been scientifically evaluated, and therefore, little is known about their adverse effects, compromising the use of these products in folk medicine (Wiesner, 2014). In this context, an increment in researches to confirm the efficacy and mainly the safety of natural products used by population is needed (Agbaire et al., 2013).

An example of natural product with little information in the literature is Piper cernuum Vell (Piperaceae), popularly known as Pariparoba, a native species from the Atlantic rainforest (Mariot et al., 2002). This species is considered a medicinal plant, and is commonly used by rural and urban inhabitants of Region of Vale do Ribeira, State of São Paulo, Brazil to treat pain conditions, such as bellyache and muscle pain (topically), besides hepatic and renal complications (Di Stasi et al., 2002).

Pharmacologically, there is a lack of information about the P. cernuum's leaves extract effects, just few works showing its antileishmanial (Capello et al., 2015) and antimicrobial (Costantin et al., 2001) effects. Similarly, the literature is scarce of information about the P. cernuum safety profile. It is known that this medicinal plant is popularly used, therefore we emphasize the data presented in this study about acute and sub-acute toxicity, genotoxicity and cytotoxicity of P. cernuum leaves extract.

Section snippets

Plant material and preparation of extract

P. cernuum Vell (Piperaceae) leaves were collected (a cultivated specie) in Blumenau, a city in the state of Santa Catarina, Brazil (Latitude: 26°58'43.8"S, Longitude: 49°03'43.0"W, and Altitude: 200 m) in March (summer) 2015. Prof. Andre Luis de Gasper, Curator Herbarium of Dr. Roberto Miguel Klein from Universidade Regional de Blumenau, classified the material. Samples of the specimen were deposited in this Herbarium with the number 41606.

The leaves were dried at 40 °C for seven days,

Results and discussion

As previously mentioned, P. cernuum leaves is used in the folk medicine as infusion or macerate with alcohol to treat pain conditions, and renal and hepatic complications (Mariot et al., 2002, Di Stasi et al., 2002). However, there is few data about its pharmacological and toxicological effects. In this context, we investigated the acute and sub-acute toxicity, mutagenic, genotoxicity and cytotoxicity profile of P. cernuum hydroalcoholic extract using in vitro and in vivo experimental models.

Conclusion

Our data demonstrates for the first time that P. cernuum leaves extract is relative safe to acute and sub-acute use. However, it is important to point out that extract presents genotoxic effects in vitro only in high concentrations. In this way, further studies are required to evaluate the safety of long-term exposure, as well as the genotoxic effects in vivo.

Acknowledgements

This study was supported by government grants from Conselho Nacional de Desenvolvimento Científico e Tecnológico, Brazil (CNPq, process n. 444682/2014-7) and Fundação de Amparo a Pesquisa e Inovação do Estado de Santa Catarina (FAPESC), Brazil. Universidade do Vale do Itajaí (UNIVALI), Brazil, also provided financial support.

Conflict of interest

The authors declare no conflict of interests.

Author's contributions

J.R. Santin, J.C. Vitorino, and A. Malheiros conceived and designed the study; F.R. Wolff, M.F. Broering, J.D. Jurcevic and T. Zermiani performed the experiments, analyzed the data. J.R. Santin and A. Malheiros wrote the paper; A. Malheiros and F.R. Wolff collected the plant and carried out the extraction. All authors read and approved the final manuscript.

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