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Geigerin-induced cytotoxicity in a murine myoblast cell line (C2C12)

Geigeria poisoning in sheep, locally known as ‘vermeersiekte’, is an economically important plant poisoning in southern Africa. The toxic principles contained by the toxic plants are believed to be several sesquiterpene lactones, such as geigerin, vermeeric acid and vermeerin, which cause striated m...

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Autores principales: Botha, Christo J., Clift, Sarah J., Ferreira, Gezina C.H., Masango, Mxolisi G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: AOSIS 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6238781/
https://www.ncbi.nlm.nih.gov/pubmed/29113440
http://dx.doi.org/10.4102/ojvr.v84i1.1465
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author Botha, Christo J.
Clift, Sarah J.
Ferreira, Gezina C.H.
Masango, Mxolisi G.
author_facet Botha, Christo J.
Clift, Sarah J.
Ferreira, Gezina C.H.
Masango, Mxolisi G.
author_sort Botha, Christo J.
collection PubMed
description Geigeria poisoning in sheep, locally known as ‘vermeersiekte’, is an economically important plant poisoning in southern Africa. The toxic principles contained by the toxic plants are believed to be several sesquiterpene lactones, such as geigerin, vermeeric acid and vermeerin, which cause striated muscle lesions in small stock. Because of ethical issues surrounding the use of live animals in toxicity studies, there is currently a dire need to establish an in vitro model that can be used to replace traditional animal experimentation. The objective of this study was to determine the cytotoxicity of geigerin in a murine myoblast cell line (C2C12) using methyl-thiazol-tetrazolium (MTT) and lactate dehydrogenase (LDH) assays, annexin V and propidium iodide (PI) flow cytometry and transmission electron microscopy (TEM). Mouse myoblasts were exposed to 2.0 mM, 2.5 mM and 5.0 mM geigerin for 24, 48 and 72 h. A concentration-dependent cytotoxic response was observed. Apoptosis was detected by means of annexin V flow cytometry during the first 24 h and apoptotic bodies were also visible on TEM. According to the LDH and PI flow cytometry results, myoblast cell membranes were not injured. We concluded that the murine myoblast cell line (C2C12) is a suitable model for future studies planned to evaluate the cytotoxicity of other and combinations of sesquiterpene lactones, with and without metabolic activation, implicated in ‘vermeersiekte’ and to elucidate the subcellular effects of these myotoxins on cultured myoblasts.
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spelling pubmed-62387812018-11-26 Geigerin-induced cytotoxicity in a murine myoblast cell line (C2C12) Botha, Christo J. Clift, Sarah J. Ferreira, Gezina C.H. Masango, Mxolisi G. Onderstepoort J Vet Res Original Research Geigeria poisoning in sheep, locally known as ‘vermeersiekte’, is an economically important plant poisoning in southern Africa. The toxic principles contained by the toxic plants are believed to be several sesquiterpene lactones, such as geigerin, vermeeric acid and vermeerin, which cause striated muscle lesions in small stock. Because of ethical issues surrounding the use of live animals in toxicity studies, there is currently a dire need to establish an in vitro model that can be used to replace traditional animal experimentation. The objective of this study was to determine the cytotoxicity of geigerin in a murine myoblast cell line (C2C12) using methyl-thiazol-tetrazolium (MTT) and lactate dehydrogenase (LDH) assays, annexin V and propidium iodide (PI) flow cytometry and transmission electron microscopy (TEM). Mouse myoblasts were exposed to 2.0 mM, 2.5 mM and 5.0 mM geigerin for 24, 48 and 72 h. A concentration-dependent cytotoxic response was observed. Apoptosis was detected by means of annexin V flow cytometry during the first 24 h and apoptotic bodies were also visible on TEM. According to the LDH and PI flow cytometry results, myoblast cell membranes were not injured. We concluded that the murine myoblast cell line (C2C12) is a suitable model for future studies planned to evaluate the cytotoxicity of other and combinations of sesquiterpene lactones, with and without metabolic activation, implicated in ‘vermeersiekte’ and to elucidate the subcellular effects of these myotoxins on cultured myoblasts. AOSIS 2017-10-31 /pmc/articles/PMC6238781/ /pubmed/29113440 http://dx.doi.org/10.4102/ojvr.v84i1.1465 Text en © 2017. The Authors https://creativecommons.org/licenses/by/4.0/ Licensee: AOSIS. This work is licensed under the Creative Commons Attribution License.
spellingShingle Original Research
Botha, Christo J.
Clift, Sarah J.
Ferreira, Gezina C.H.
Masango, Mxolisi G.
Geigerin-induced cytotoxicity in a murine myoblast cell line (C2C12)
title Geigerin-induced cytotoxicity in a murine myoblast cell line (C2C12)
title_full Geigerin-induced cytotoxicity in a murine myoblast cell line (C2C12)
title_fullStr Geigerin-induced cytotoxicity in a murine myoblast cell line (C2C12)
title_full_unstemmed Geigerin-induced cytotoxicity in a murine myoblast cell line (C2C12)
title_short Geigerin-induced cytotoxicity in a murine myoblast cell line (C2C12)
title_sort geigerin-induced cytotoxicity in a murine myoblast cell line (c2c12)
topic Original Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6238781/
https://www.ncbi.nlm.nih.gov/pubmed/29113440
http://dx.doi.org/10.4102/ojvr.v84i1.1465
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