背景:光生物调节在减轻Bothrops蛇咬伤引起的局部影响方面表现出了希望;但是,这种保护的潜在机制尚未完全理解。在这里,光生物调节效应对C2C12成肌细胞再生反应的影响暴露于Bithropsjaracussu毒液(BjsuV),以及所涉及的机制进行了调查。
结果:将C2C12成肌细胞暴露于BjsuV(12.5μg/mL),并用660nm(14.08mW;0.04cm2;352mW/cm2)或780nm(17.6mW;0.04cm2;440mW/cm2)的激光照射10秒钟,以提供3.52和4.4J/cm2的能量密度,总能量为0.1408和0.176J,分别。通过伤口愈合测定评估细胞迁移。MAPKp38-α的表达,NF-█β,Myf5Pax-7MyoD,和肌细胞生成素蛋白通过蛋白质印迹分析进行评估。此外,白细胞介素IL1-β,通过ELISA测量上清液中的IL-6、TNF-α和IL-10水平。PBM应用于C2C12细胞暴露于BjsuV促进细胞迁移,增加肌源性因子(Pax7,MyF5,MyoD和肌原蛋白)的表达,降低了促炎细胞因子的水平,IL1-β,IL-6,TNF-α,并增加抗炎细胞因子IL-10的水平。此外,PBM下调NF-kB的表达,对p38MAKP无影响。
结论:这些数据表明,PBM对肌细胞的保护似乎与肌源性因子的增加以及炎症介质的调节有关。PBM治疗可能提供一种新的治疗策略,通过促进肌肉再生和减少炎症过程来解决蛇咬伤引起的局部效应。
BACKGROUND: Photobiomodulation has exhibited promise in mitigating the local effects induced by Bothrops snakebite envenoming; however, the mechanisms underlying this protection are not yet fully understood. Herein, the effectiveness of photobiomodulation effects on regenerative response of C2C12 myoblast cells following exposure to Bothrops jararacussu venom (BjsuV), as well as the mechanisms involved was investigated.
RESULTS: C2C12 myoblast cells were exposed to BjsuV (12.5 μg/mL) and irradiated once for 10 seconds with laser light of 660 nm (14.08 mW; 0.04 cm2; 352 mW/cm2) or 780 nm (17.6 mW; 0.04 cm2; 440 mW/ cm2) to provide energy densities of 3.52 and 4.4 J/cm2, and total energies of 0.1408 and 0.176 J, respectively. Cell migration was assessed through a wound-healing assay. The expression of MAPK p38-α, NF-Кβ, Myf5, Pax-7, MyoD, and myogenin proteins were assessed by western blotting analysis. In addition, interleukin IL1-β, IL-6, TNF-alfa and IL-10 levels were measured in the supernatant by ELISA. The PBM applied to C2C12 cells exposed to BjsuV promoted cell migration, increase the expression of myogenic factors (Pax7, MyF5, MyoD and myogenin), reduced the levels of proinflammatory cytokines, IL1-β, IL-6, TNF-alfa, and increased the levels of anti-inflammatory cytokine IL-10. In addition, PBM downregulates the expression of NF-kB, and had no effect on p38 MAKP.
CONCLUSIONS: These data demonstrated that protection of the muscle cell by PBM seems to be related to the increase of myogenic factors as well as the modulation of inflammatory mediators. PBM therapy may offer a new therapeutic strategy to address the local effects of snakebite envenoming by promoting muscle regeneration and reducing the inflammatory process.