Original Research

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

Christo J. Botha, Sarah J. Clift, Gezina C.H. Ferreira, Mxolisi G. Masango
Onderstepoort Journal of Veterinary Research | Vol 84, No 1 | a1465 | DOI: https://doi.org/10.4102/ojvr.v84i1.1465 | © 2017 Christo J. Botha, Sarah J. Clift, Gezina C.H. Ferreira, Mxolisi G. Masango | This work is licensed under CC Attribution-NoDerivatives 4.0
Submitted: 05 May 2017 | Published: 31 October 2017

About the author(s)

Christo J. Botha, Department of Paraclinical Sciences, University of Pretoria, South Africa
Sarah J. Clift, Department of Paraclinical Sciences, University of Pretoria, South Africa
Gezina C.H. Ferreira, Department of Paraclinical Sciences, University of Pretoria, South Africa
Mxolisi G. Masango, Food, Feed and Veterinary Public Health, Agricultural Research Council-Onderstepoort Veterinary Institute, South Africa


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Abstract

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.

Keywords

Cytotoxicity; Geigeria; Myoblasts; Sesquiterpene lactones; Vermeersiekte

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