An Da Hyeon, Lee Chan Ho, Kwon Yeeun, Kim Tae Hee, Kim Eun Ji, Jung Jae In, Min Sangil, Cheong Eun Ju, Kim Sohyun, Kim Hee Kyu, Choi Sun Eun
Department of Forest Biomaterials Engineering, Kangwon National University, Chuncheon 24341, Gangwon State, Republic of Korea.
Dr.Oregonin Inc., #802 Bodeum Hall, Kangwondaehakgil 1, Chuncheon 24341, Gangwon State, Republic of Korea.
Pharmaceuticals (Basel). 2024 Dec 10;17(12):1661. doi: 10.3390/ph17121661.
Sarcopenia is characterized by the loss of muscle mass and function, increases in mortality rate, and risk of comorbidities in the elderly. This study evaluated the effects of hot water extract (AJHW) and its active compound, oregonin, on muscle atrophy and apoptosis in vitro. AJHW underwent phytochemical analysis. C2C12 cells were subjected to HO and dexamethasone to induce oxidative stress and muscle loss, after which AJHW and oregonin were administered to assess their impacts on cell viability, apoptosis, muscle protein synthesis stimulation, and muscle protein degradation inhibition. Cell viability was assessed via an MTT assay, and apoptosis was analyzed by measuring Bcl-2, Bax, cleaved caspase-3, and cleaved PARP through Western blotting. Western blotting and RT-PCR were utilized to analyze MyoD, Myogenin, Atrogin-1, and MuRF1 protein and gene expression in a muscle atrophy model, as well as the Akt/mTOR and FoxO3α pathways. AJHW was confirmed to contain oregonin, an active compound. AJHW and oregonin significantly increased cell viability and reduced apoptosis by upregulating Bcl-2 and downregulating Bax, cleaved caspase-3, and cleaved PARP. They significantly enhanced muscle protein synthesis through the upregulation of MyoD and Myogenin, while diminishing muscle degradation by downregulating Atrogin-1 and MuRF1. The activation of the Akt/mTOR pathway and inhibition of the FoxO3α pathway were also observed. AJHW and oregonin effectively prevented muscle cell apoptosis, promoted muscle protein synthesis, and inhibited muscle protein degradation in vitro. These results suggest that AJHW and oregonin could serve as therapeutic agents to prevent and treat sarcopenia.
肌肉减少症的特征是肌肉质量和功能丧失、死亡率增加以及老年人患合并症的风险增加。本研究评估了热水提取物（AJHW）及其活性化合物奥勒冈因对体外肌肉萎缩和细胞凋亡的影响。对AJHW进行了植物化学分析。将C2C12细胞暴露于过氧化氢和地塞米松以诱导氧化应激和肌肉损失，之后给予AJHW和奥勒冈因以评估它们对细胞活力、细胞凋亡、肌肉蛋白合成刺激和肌肉蛋白降解抑制的影响。通过MTT法评估细胞活力，并通过蛋白质印迹法测量Bcl-2、Bax、裂解的半胱天冬酶-3和裂解的聚（ADP-核糖）聚合酶来分析细胞凋亡。利用蛋白质印迹法和逆转录-聚合酶链反应分析肌肉萎缩模型中MyoD、肌细胞生成素、肌肉萎缩相关蛋白-1和肌肉特异性泛素连接酶1的蛋白质和基因表达，以及Akt/mTOR和叉头框蛋白O3α途径。证实AJHW含有活性化合物奥勒冈因。AJHW和奥勒冈因通过上调Bcl-2和下调Bax、裂解的半胱天冬酶-3和裂解的聚（ADP-核糖）聚合酶，显著提高细胞活力并减少细胞凋亡。它们通过上调MyoD和肌细胞生成素显著增强肌肉蛋白合成，同时通过下调肌肉萎缩相关蛋白-1和肌肉特异性泛素连接酶1减少肌肉降解。还观察到Akt/mTOR途径的激活和叉头框蛋白O3α途径的抑制。AJHW和奥勒冈因在体外有效预防肌肉细胞凋亡，促进肌肉蛋白合成并抑制肌肉蛋白降解。这些结果表明，AJHW和奥勒冈因可作为预防和治疗肌肉减少症的治疗药物。
