背景:心血管疾病是全球发病率和死亡率的主要原因。缺血性心脏病是对细胞结构和功能最有害的疾病之一。再灌注治疗后,一系列副作用变得明显,包括改变的细胞活力,氧化应激增强,激活自噬,和增加细胞凋亡。光生物调节(PBM)已用于心脏缺氧的实验模型中,以增强线粒体反应并改善受损组织的生化变化。然而,PBM对缺氧/复氧培养心肌细胞的影响尚未完全确定。
方法:将H9C2心肌细胞暴露于浓度为300μMCoCl2的缺氧中24小时,然后通过在常氧培养基中孵育16小时的复氧。在缺氧后使用GaAlAs激光(850nm)以lJ/cm2的强度进行处理。将细胞分为三组:CT组(在常氧条件下维持的细胞),HR组(细胞维持在缺氧和复氧条件下,不治疗),HR+PBM组(在经历PBM处理的缺氧和复氧条件下维持的细胞)。使用MTT分析细胞活力,通过蛋白质印迹评估蛋白质表达。使用具有Tukey事后检验的单向ANOVA进行数据分析。当p<0.05时,差异显著。
结果:PBM在1J/cm2的强度下减轻了由缺氧/复氧引起的细胞存活的改变。此外,它显着增加了Nrf2,HSP70,mTOR,LC3II,LC3II/I,和Caspase-9,同时降低PGC-1α的表达,SOD2,黄嘌呤氧化酶,Beclin-1LC3I,还有Bax.
结论:PBM在1J/cm2的强度下逆转了与氧化应激相关的变化,线粒体生物发生,自噬,以及心肌细胞培养物中缺氧和复氧引起的细胞凋亡。
BACKGROUND: Cardiovascular diseases are the leading cause of morbidity and mortality worldwide. Ischemic heart disease is one of the most harmful conditions to cellular structure and function. After reperfusion treatment, a spectrum of adverse effects becomes evident, encompassing altered cell viability, heightened oxidative stress, activated autophagy, and increased apoptosis. Photobiomodulation (PBM) has been utilized in experimental models of cardiac hypoxia to enhance mitochondrial response and ameliorate biochemical changes in injured tissue. However, the effects of PBM on cultured cardiomyocytes subjected to hypoxia/reoxygenation are not yet well established.
METHODS: H9C2 cardiomyocytes were exposed to hypoxia with concentrations of 300 μM CoCl2 for 24 h, followed by 16 h of reoxygenation through incubation in a normoxic medium. Treatment was conducted using GaAIAs Laser (850 nm) after hypoxia at an intensity of 1 J/cm2. Cells were divided into three groups: Group CT (cells maintained under normoxic conditions), Group HR (cells maintained in hypoxia and reoxygenation conditions without treatment), Group HR + PBM (cells maintained in hypoxia and reoxygenation conditions that underwent PBM treatment). Cell viability was analyzed using MTT, and protein expression was assessed by western blot. One-way ANOVA with the Tukey post hoc test was used for data analysis. Differences were significant when p < 0.05.
RESULTS: PBM at an intensity of 1 J/cm2 mitigated the alterations in cell survival caused by hypoxia/reoxygenation. Additionally, it significantly increased the expression of proteins Nrf2, HSP70, mTOR, LC3II, LC3II/I, and Caspase-9, while reducing the expression of PGC-1α, SOD2, xanthine oxidase, Beclin-1, LC3I, and Bax.
CONCLUSIONS: PBM at intensities of 1 J/cm2 reverses the changes related to oxidative stress, mitochondrial biogenesis, autophagy, and apoptosis caused by hypoxia and reoxygenation in a culture of cardiomyocytes.