Wang Keke, Zeng Hao, Yang Hua
State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing, 210009, China.
Chin Med. 2024 Dec 31;19(1):180. doi: 10.1186/s13020-024-01034-5.
Lovastatin, the main lipid-lowering component in red yeast rice, is a golden anti-lipid drug, but its long-term application is continuously challenged by potential skeletal muscle atrophy. Daidzein, an isoflavone derived from soybeans and many Chinese medicines, shows therapeutic potential in treating muscle-related diseases and metabolic disorders. However, whether daidzein can improve lovastatin-induced muscle atrophy and the specific mechanism needs to further study.
Lovastatin-induced mice and zebrafish muscle atrophy models were used to validate the protective effect of daidzein in vivo. And the lovastatin-induced C2C12 myotube atrophy model was employed to validate the therapeutic efficacy and investigate the specific mechanism of daidzein in vitro. We combined specific siRNA targeting FOXO3a and AMPK-selective inhibitor, agonist to elucidate AMPK/FOXO3a-dependent muscle-protective mechanism of daidzein. The anti-atrophy effects of daidzein through blockage of abnormal activation of AMPK/FOXO3a was presented in Immunofluorescence, H&E staining, Western blot, qRT-PCR. Serum creatine kinase level was detected by ELISA and we used mouse muscle grip instrument to detect the strength of mouse muscles.
In this study, we demonstrated that daidzein could dose-dependently alleviate lovastatin-induced mice skeletal muscle atrophy, reduce serum creatine kinase, and improve muscle grip strength in mice. Mechanistically, daidzein inhibited lovastatin-induced FOXO3a phosphorylation caused by AMPK activation, thereby inhibiting FOXO3a nuclear translocation to restrain the expression of muscle-related proteins Atrogin-1 and MuRF-1. In C2C12 myotube, administration of AMPK-selective inhibitor Compound C recapitulated the therapeutic effects of daidzein against lovastatin-induced myotubes atrophy, while the anti-atrophy effects of daidzein were lost in the presence of AMPK-selective agonist MK-3903. In lovastatin-induced mice muscle atrophy models, Compound C elicited similar anti-atrophy effects as daidzein, but this effect was not potentiated when it was applied in combination with daidzein, suggesting that daidzein exerted therapeutic efficacy dependent on blockage of AMPK activity.
Our study identified daidzein as an effective component that ameliorated lovastatin-induced skeletal muscle atrophy through blockage of abnormal activation of AMPK/FOXO3a and transcriptional activation of genes encoding downstream muscle-related proteins. Our results also highlighted the therapeutic potential by regulating the AMPK/FOXO3a axis in management of statin-induced myotoxicity.
洛伐他汀是红曲中的主要降脂成分，是一种黄金降脂药物，但其长期应用不断受到潜在骨骼肌萎缩的挑战。大豆苷元是一种来源于大豆和多种中药的异黄酮，在治疗肌肉相关疾病和代谢紊乱方面显示出治疗潜力。然而，大豆苷元是否能改善洛伐他汀引起的肌肉萎缩及其具体机制尚需进一步研究。
采用洛伐他汀诱导的小鼠和斑马鱼肌肉萎缩模型在体内验证大豆苷元的保护作用。并采用洛伐他汀诱导的C2C12肌管萎缩模型在体外验证大豆苷元的治疗效果并研究其具体机制。我们联合靶向FOXO3a的特异性siRNA和AMPK选择性抑制剂、激动剂来阐明大豆苷元的AMPK/FOXO3a依赖性肌肉保护机制。通过免疫荧光、苏木精-伊红染色、蛋白质免疫印迹、实时定量聚合酶链反应展示了大豆苷元通过阻断AMPK/FOXO3a的异常激活产生的抗萎缩作用。通过酶联免疫吸附测定法检测血清肌酸激酶水平，并使用小鼠肌肉握力仪检测小鼠肌肉力量。
在本研究中，我们证明大豆苷元可以剂量依赖性地减轻洛伐他汀诱导的小鼠骨骼肌萎缩，降低血清肌酸激酶，并提高小鼠的肌肉握力。机制上，大豆苷元抑制了由AMPK激活引起的洛伐他汀诱导的FOXO3a磷酸化，从而抑制FOXO3a核转位，以抑制肌肉相关蛋白Atrogin-1和MuRF-1的表达。在C2C12肌管中，给予AMPK选择性抑制剂化合物C重现了大豆苷元对洛伐他汀诱导的肌管萎缩的治疗效果，而在存在AMPK选择性激动剂MK-3903的情况下，大豆苷元的抗萎缩作用消失。在洛伐他汀诱导的小鼠肌肉萎缩模型中，化合物C产生了与大豆苷元相似的抗萎缩作用，但与大豆苷元联合应用时这种作用并未增强，这表明大豆苷元发挥治疗效果依赖于阻断AMPK活性。
我们的研究确定大豆苷元是一种有效的成分，它通过阻断AMPK/FOXO3a的异常激活和编码下游肌肉相关蛋白的基因的转录激活来改善洛伐他汀诱导的骨骼肌萎缩。我们的结果还突出了通过调节AMPK/FOXO3a轴在他汀类药物诱导的肌毒性管理中的治疗潜力。
