Department of Traditional Chinese Medicine, Shenzhen Hospital, Southern Medical University, No. 1333, Xinhu Road, Bao 'an District, Shenzhen, Guangdong, 518000, China; School of Traditional Chinese Medicine, Southern Medical University, No. 1023-1063, Shatai South Road, Baiyun District, Guangzhou, Guangdong, 510000, China.
Department of Traditional Chinese Medicine, Shenzhen Hospital, Southern Medical University, No. 1333, Xinhu Road, Bao 'an District, Shenzhen, Guangdong, 518000, China.
J Ethnopharmacol. 2023 Dec 5;317:116737. doi: 10.1016/j.jep.2023.116737. Epub 2023 Jun 8.
The combination of Astragulus embranaceus (Fisch.) Bge (Huangqi) and Dioscorea opposita Thunb (Shanyao) are one of the most widely accepted herb pairs in traditional Chinese medicine prescriptions for treating sarcopenia. However, the mechanisms underlying the combination of these herbs for anti-sarcopenia treatment are not yet fully understood.
To investigate the potential effect of the Astragulus embranaceus (Fisch.) Bge and Dioscorea opposita Thunb herb pair (Ast-Dio) on sarcopenia in mice that have been induced with senile type 2 diabetes mellitus, as well as to explore the underlying mechanisms related to the Rab5a/mTOR signaling pathway and mitochondrial quality control.
Network pharmacology was utilized to identify the main active ingredients of Ast-Dio and potential therapeutic targets for sarcopenia. Gene Ontology function and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were conducted to explore the underlying mechanisms of Ast-Dio in treating sarcopenia. The high-performance liquid chromatography method coupled with triple-quadrupole tandem mass spectrometry was developed to quantify the major constituents of Ast-Dio. Male C57/BL6 mice, aged 12 months, induced with type 2 diabetes mellitus via streptozotocin were divided into three groups for 8 weeks: the model group, Ast-Dio treatment group (7.8 g/kg), and metformin treatment group (100 mg/kg). Normal control groups included mice aged 3 and 12 months, respectively. The study monitored changes in fasting blood glucose levels, grip strength, and body weight during 8 weeks of intragastric administration. Liver and kidney function in mice was evaluated by measuring the levels of serum creatinine, alanine transaminase, and aspartate transaminase. Skeletal muscle mass condition was evaluated by muscle weight, and hematoxylin and eosin staining. Protein and mRNA expressions related to muscle atrophy, mitochondrial quality control, and the Rab5a/mTOR signaling pathway were detected using immunofluorescence staining, immunohistochemical staining, Western blotting, and quantitative real-time polymerase chain reaction. In addition, transmission electron microscopy was employed to investigate the condition of mitochondria in the groups.
Through the prediction analysis of network pharmacology, we identified mTOR as one of the primary targets for Ast-Dio therapy of sarcopenia. Gene Ontology functional enrichment analysis revealed that mitochondrial control quality is crucial in the treatment of sarcopenia with Ast-Dio. Our findings showed that senile type 2 diabetes mellitus induced muscle mass loss and a reduction in grip strength, both of which were dramatically restored by Ast-Dio treatment. Notably, Ast-Dio increased Myogenin expression while decreasing Atrogin-1 and MuRF-1 expression. Additionally, Ast-Dio activated Rab5a/mTOR and its downstream effector AMPK. Moreover, Ast-Dio modulated mitochondrial quality control by decreasing Mitofusin-2 expression while increasing the expression of TFAM, PGC-1α, and MFF.
Our results suggest that Ast-Dio treatment may alleviate sarcopenia in mice with senile type 2 diabetes mellitus through its effects on the Rab5a/mTOR pathway and mitochondrial quality control.
黄芪（Huangqi）和山药（Shanyao）的组合是中药方剂中治疗肌肉减少症最广泛接受的草药组合之一。然而，这种草药组合治疗肌肉减少症的机制尚不完全清楚。
研究黄芪（Astragulus embranaceus (Fisch.) Bge）和山药（Dioscorea opposita Thunb）草药组合（Ast-Dio）对衰老型 2 型糖尿病诱导的肌肉减少症小鼠的潜在影响，并探讨与 Rab5a/mTOR 信号通路和线粒体质量控制相关的潜在机制。
利用网络药理学方法鉴定 Ast-Dio 的主要活性成分和治疗肌肉减少症的潜在治疗靶点。通过基因本体论功能和京都基因与基因组百科全书途径富集分析探讨 Ast-Dio 治疗肌肉减少症的潜在机制。采用高效液相色谱法结合三重四极杆串联质谱法开发了一种用于定量分析 Ast-Dio 主要成分的方法。将 12 个月大的雄性 C57/BL6 小鼠通过链脲佐菌素诱导 2 型糖尿病，分为 3 组，进行 8 周的治疗：模型组、Ast-Dio 治疗组（7.8 g/kg）和二甲双胍治疗组（100 mg/kg）。正常对照组包括 3 个月和 12 个月大的小鼠。研究监测了 8 周灌胃期间空腹血糖水平、握力和体重的变化。通过测量血清肌酐、丙氨酸转氨酶和天冬氨酸转氨酶水平来评估小鼠的肝肾功能。通过肌肉重量和苏木精-伊红染色评估骨骼肌状况。使用免疫荧光染色、免疫组织化学染色、Western blot 和实时定量聚合酶链反应检测与肌肉萎缩、线粒体质量控制和 Rab5a/mTOR 信号通路相关的蛋白和 mRNA 表达。此外，还通过透射电子显微镜观察了各组线粒体的情况。
通过网络药理学的预测分析，我们确定 mTOR 是 Ast-Dio 治疗肌肉减少症的主要靶点之一。基因本体论功能富集分析表明，线粒体质量控制对 Ast-Dio 治疗肌肉减少症至关重要。我们的研究结果表明，衰老型 2 型糖尿病诱导肌肉质量减少和握力下降，而 Ast-Dio 治疗可显著恢复这些症状。值得注意的是，Ast-Dio 增加了 Myogenin 的表达，同时降低了 Atrogin-1 和 MuRF-1 的表达。此外，Ast-Dio 激活了 Rab5a/mTOR 及其下游效应物 AMPK。此外，Ast-Dio 通过降低 Mitofusin-2 的表达和增加 TFAM、PGC-1α 和 MFF 的表达来调节线粒体质量控制。
我们的结果表明，Ast-Dio 治疗可能通过其对 Rab5a/mTOR 通路和线粒体质量控制的影响，缓解衰老型 2 型糖尿病小鼠的肌肉减少症。
