结直肠癌引起成肌细胞恶病质的体外模型研究

Research on in vitro models of cachexia in myoblast cells caused by colorectal cancer

目的 恶病质作为一种全身消耗性疾病,其主要特点为难以逆转的全身骨骼肌质量减少,严重损害患者健康状态。进一步解析恶病质引起骨骼肌消耗的机制以探寻潜在治疗靶点是改善恶病质患者生存质量的重要策略。方法 利用Incucyte持续观察肿瘤细胞上清对成肌细胞增殖、迁移能力的影响,并利用流式细胞术检测成肌细胞凋亡水平改变。通过RT-qPCR检测分析肿瘤细胞上清对体外诱导分化肌管的成肌分化、肌肉萎缩及脂质过氧化等相关基因的表达。利用细胞成像和流式细胞术共同检测肿瘤细胞上清诱导的成肌细胞线粒体损伤情况。结果 通过体外模拟恶病质对成肌细胞和肌管的影响,观察到在肿瘤上清刺激下,成肌细胞迁移能力减弱、细胞凋亡加剧。且诱导分化后的成肌相关基因表达下调、肌肉萎缩相关基因表达上调,同时伴随脂质过氧化,进一步研究发现成肌细胞线粒体膜电位出现了明显变化。结论 恶病质可通过诱导脂质过氧化和线粒体损伤造成骨骼肌萎缩,保护线粒体不受恶病质因子的损伤可能是治疗恶病质骨骼肌减少的潜在策略。

Objective Cachexia,as a systemic wasting disease,is characterized by an irreversible loss of skeletal muscle mass,which seriously impairs the health of patients.Further understanding the mechanism of skeletal muscle wasting caused by cachexia and exploring potential therapeutic targets are important strategies to improve the quality of life of cachexia patients.Methods Incucyte was used to continuously observe the effects of tumor cell supernatant on the proliferation and migration of myoblasts and flow cytometry was used to detect the changes of apoptosis of myoblasts.RT-qPCR was used to detect the expression of myogenic differentiation,muscle atrophy,and lipid peroxidation-related genes in the tumor cell supernatant.Cell imaging and flow cytometry were used to detect mitochondrial damage in myoblasts induced by tumor cell supernatant.Results By simulating the effect of cachexia on myoblasts and myotubes in vitro,it was observed that the migration ability of myoblasts was weakened and apoptosis was aggravated under the stimulation of tumor supernatant.After induction,the expression of myogenic genes was down-regulated,and the expression of muscle atrophy-related genes was up-regulated,accompanied by lipid peroxidation.Further studies showed that the mitochondrial membrane potential of myoblasts changed significantly.Conclusion Cachexia can cause skeletal muscle atrophy by inducing lipid peroxidation and mitochondrial damage.Protecting mitochondria from cachexia factors may be a potential strategy for the treatment of skeletal muscle loss in cachexia.