Efeito do laser de baixa intensidade sobre células musculares submetidas à ação do veneno de serpentes do gênero bothrops: mecanismo de morte celular

Abstract

The envenoming caused by snakes of the Bothrops genus is considered a neglected tropical disease and is frequently associated with severe and complex local manifestations; such as edema, intense pain, hemorrhage and myonecrosis. The specific and conventional treatment for this condition is the use of the anti-venom serum, which neutralizes the majority of the circulating venom, thus minimizing its systemic effects. However, its local action is reduced and, therefore, the patient may present permanent tissue damage and, in some cases, even amputation of the affected member. Thus, therapies are sought that can complement the action of the serum and minimize the local effects of the venom. Previous studies in our group have demonstrated that the low level laser (LLL) increases the viability of C2C12 muscle cells incubated with the venom of Bothrops jararacussu and further promotes the differentiation of these cells. Thus, the objective of this work was to evaluate the effect of LLL on C2C12 muscle cells submitted to B. jararaca, B. jararacussu and B. moojeni venoms for their mechanism of cell death (necrosis and apoptosis). To do so, the cells were incubated with the respective venoms and immediately irradiated with LLL at the wavelength of 660 nm, energy density of 2.5 J/cm2, potency of 10 mW, area of 0.045 cm2 and time of application of 20 seconds. The cells were incubated for 2 hours and then flow cytometry was performed for analysis of cell death. Cell viability was also evaluated along with the dosage of inflammatory mediators such as interleukins IL-1 β, IL-6, IL-10 and Tumor Necrosis Factor α (TNF-α). Our results showed that the application of LLL decreases cell death in the different venoms by necrosis and apoptosis; increases cell viability; decreases the release of IL-6 to all venoms and IL-1 β for the venoms of B. jararaca and B. moojeni, but does not influence the release of IL-10 and TNF-α. Thus, the LLL protects the muscle cells against the action of the venoms studied; increasing cell viability by decreasing cell death (necrosis and apoptosis) and minimizing the inflammatory process.