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Autophagy induced by purple pitanga (Eugenia uniflora L.) extract triggered a cooperative effect on inducing the hepatic stellate cell death

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Abstract

Activated hepatic stellate cells (HSC) are the major source of collagen I in liver fibrosis. Eugenia uniflora L. is a tree species that is widely distributed in South America. E. uniflora L. fruit—popularly known as pitanga—has been shown to exert beneficial properties. Autophagy contributes to the maintenance of cellular homeostasis and survival under stress situation, but it has also been suggested to be an alternative cell death pathway. Mitochondria play a pivotal role on signaling cell death. Mitophagy of damaged mitochondria is an important cell defense mechanism against organelle-mediated cell death signaling. We previously found that purple pitanga extract induced mitochondrial dysfunction, cell cycle arrest, and death by apoptosis and necrosis in GRX cells, a well-established activated HSC line. We evaluated the effects of 72-h treatment with crescent concentrations of purple pitanga extract (5 to 100 μg/mL) on triggering autophagy in GRX cells, as this is an important mechanism to cells under cytotoxic conditions. We found that all treated cells presented an increase in the mRNA expression of autophagy-related protein 7 (ATG7). Concomitantly, flow cytometry and ultrastructural analysis of treated cells revealed an increase of autophagosomes/autolysosomes that consequentially led to an increased mitophagy. As purple pitanga extract was previously found to be broadly cytotoxic to GRX cells, we postulated that autophagy contributes to this scenario, where cell death seems to be an inevitable fate. Altogether, the effectiveness on inducing activated HSC death can make purple pitanga extract a good candidate on treating liver fibrosis.

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Acknowledgments

The authors acknowledge Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), and Fundação de Amparo à Pesquisa do Rio Grande do Sul (FAPERGS) for financial support; and Embrapa Clima Temperado for their collaboration and supply of fruit samples. The authors thank to Centro de Microscopia e Microanálise (CMM-UFRGS) by technical assistance in confocal and transmission electron microscopy.

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Authors and Affiliations

  1. Programa de Pós-Graduação em Ciências Biológicas-Bioquímica, ICBS, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brasil

    Cristiane C. Denardin, Leo A. M. Martins, Mariana M. Parisi, Moema Queiroz Vieira, Silvia R. Terra, Florencia M. Barbé-Tuana & Fátima Costa Rodrigues Guma

  2. Curso de Farmácia, Universidade Federal do Pampa (UNIPAMPA), Campus Uruguaiana, Uruguaiana, RS, Brasil

    Cristiane C. Denardin

  3. Departamento de Bioquímica, Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos 2600-Anexo, Lab 21, CEP: 90035-003, Porto Alegre, RS, Brasil

    Fátima Costa Rodrigues Guma

  4. Núcleo Integrado de Desenvolvimento em Análises Laboratoriais (NIDAL), Departamento de Tecnologia e Ciência de Alimentos, Universidade Federal de Santa Maria, Santa Maria, RS, Brasil

    Tatiana Emanuelli

  5. Empresa Brasileira de Pesquisa Agropecuária de Clima Temperado, Pelotas, RS, Brasil

    Márcia Vizzotto

  6. Departamento de Histologia e Embriologia, ICB, UFRJ, Rio de Janeiro, RJ, Brasil

    Radovan Borojevic

  7. Centro de Microscopia e Microanálise, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brasil

    Fátima Costa Rodrigues Guma

Authors
  1. Cristiane C. Denardin
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  2. Leo A. M. Martins
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  3. Mariana M. Parisi
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  4. Moema Queiroz Vieira
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  5. Silvia R. Terra
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  6. Florencia M. Barbé-Tuana
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  7. Radovan Borojevic
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  8. Márcia Vizzotto
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  9. Tatiana Emanuelli
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  10. Fátima Costa Rodrigues Guma
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Corresponding author

Correspondence to Fátima Costa Rodrigues Guma.

Additional information

Cristiane C. Denardin and Leo A. M. Martins contributed equally.

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Denardin, C.C., Martins, L.A.M., Parisi, M.M. et al. Autophagy induced by purple pitanga (Eugenia uniflora L.) extract triggered a cooperative effect on inducing the hepatic stellate cell death. Cell Biol Toxicol 33, 197–206 (2017). https://doi.org/10.1007/s10565-016-9366-5

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  • DOI: https://doi.org/10.1007/s10565-016-9366-5

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