Photobiomodulation Protects and Promotes Differentiation of C2C12 Myoblast Cells Exposed to Snake Venom

Snakebites is a neglected disease and in Brazil is considered a serious health problem, with the majority of the snakebites caused by the genus Bothrops. Antivenom therapy and other first-aid treatments do not reverse local myonecrose which is the main sequel caused by the envenomation. Several stud...

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Published in:	PloS one Vol. 11; no. 4; p. e0152890
Main Authors:	Silva, Luciana Miato Gonçalves, Silva, Camila Aparecida Alves da, Silva, Aline da, Vieira, Rodolfo Paula, Mesquita-Ferrari, Raquel Agnelli, Cogo, José Carlos, Zamuner, Stella Regina
Format:	Journal Article
Language:	English
Published:	United States Public Library of Science 08-04-2016 Public Library of Science (PLoS)
Subjects:	Adenosine Triphosphate - metabolism Analysis Animals Antivenom Apoptosis - drug effects Apoptosis - radiation effects ATP Biology and Life Sciences Bites and stings Blotting, Western Bothrops Bothrops jararacussu Cell differentiation Cell Differentiation - drug effects Cell Differentiation - radiation effects Cell Proliferation - drug effects Cell Proliferation - radiation effects Cells, Cultured Cytotoxicity Differentiation (biology) Engineering and Technology Flux density Genetic aspects Immunoenzyme Techniques Infrared lasers Irradiated Irradiation Lasers Light therapy Low level Low-Level Light Therapy Medicine and Health Sciences Mice Myoblasts Myoblasts - drug effects Myoblasts - pathology Myoblasts - radiation effects MyoD protein Myogenin Myonecrosis Myotubes Physiological aspects Protective Agents Snake bites Snake Venoms - toxicity Snakes Studies Therapy Toxicity Venom Brazil
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Summary:	Snakebites is a neglected disease and in Brazil is considered a serious health problem, with the majority of the snakebites caused by the genus Bothrops. Antivenom therapy and other first-aid treatments do not reverse local myonecrose which is the main sequel caused by the envenomation. Several studies have shown the effectiveness of low level laser (LLL) therapy in reducing local myonecrosis induced by Bothropic venoms, however the mechanism involved in this effect is unknown. In this in vitro study, we aimed to analyze the effect of LLL irradiation against cytotoxicity induced by Bothrops jararacussu venom on myoblast C2C12 cells. C2C12 were utilized as a model target and were incubated with B. jararacussu venom (12.5 μg/mL) and immediately irradiated with LLL at wavelength of red 685 nm or infrared 830 nm with energy density of 2.0, 4.6 and 7.0 J/cm2. Effects of LLL on cellular responses of venom-induced cytotoxicity were examined, including cell viability, measurement of cell damage and intra and extracellular ATP levels, expression of myogenic regulatory factors, as well as cellular differentiation. In non-irradiated cells, the venom caused a decrease in cell viability and a massive release of LDH and CK levels indicating myonecrosis. Infrared and red laser at all energy densities were able to considerably decrease venom-induced cytotoxicity. Laser irradiation induced myoblasts to differentiate into myotubes and this effect was accompanied by up regulation of MyoD and specially myogenin. Moreover, LLL was able to reduce the extracellular while increased the intracellular ATP content after venom exposure. In addition, no difference in the intensity of cytotoxicity was shown by non-irradiated and irradiated venom. LLL irradiation caused a protective effect on C2C12 cells against the cytotoxicity caused by B. jararacussu venom and promotes differentiation of these cells by up regulation of myogenic factors. A modulatory effect of ATP synthesis may be suggested as a possible mechanism mediating cytoprotection observed under laser irradiation.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Conceived and designed the experiments: SRZ RAMF. Performed the experiments: LMGS CAAS AS. Analyzed the data: SRZ CAAS LMGS RPV. Contributed reagents/materials/analysis tools: SRZ JCC RPV. Wrote the paper: SRZ RAMF LMGS RPV. Competing Interests: The authors have declared that no competing interests exist.
ISSN:	1932-6203 1932-6203
DOI:	10.1371/journal.pone.0152890