Zhou Lan, Huang Yu-Fang, Xie Hui, Mei Xiao-Yun, Cao Jun
Basic Theory of Traditional Chinese Medicine Staff Room, Basic Medical College, Nanjing University of Traditional Chinese Medicine, Nanjing, 210029, P. R. China.
Pathological Staff Room, Basic Medical College, Nanjing University of Traditional Chinese Medicine, Nanjing, 210029, P. R. China.
J Integr Neurosci. 2020 Mar 30;19(1):89-99. doi: 10.31083/j.jin.2020.01.1226.
Denervated-dependent skeletal muscle atrophy is a disease induced by skeletal muscle associated peripheral neuro-disconnection. Its specific molecular mechanisms remain unknown. The treating for denervated-dependent skeletal muscle atrophy is applied with an herbal complex Buyang Huanwu Tang used in traditional Chinese medicine and subjected to the established denervated-dependent skeletal muscle atrophy in rat models, and the therapeutic effects and associated mechanisms were evaluated in the pathogenesis of denervated-dependent skeletal muscle atrophy. Denervated-dependent skeletal muscle atrophy in rats was established and randomly divided into eight groups, including Normal control, Model, Positive control, Model + Buyang Huanwu Tang, Model + astragalus extracts, Model + Buyang Huanwu Tang-astragalus, Buyang Huanwu Tang + LY294002, and astragalus extract + LY294002 group. Hematoxylin-eosin staining and quantitative RT-PCR (qRT-PCR) assay were used to examine the inflammatory response of muscle tissues. Quantitative RT-PCR and Western blotting assay were utilized to analyze mRNA and protein expression. Immunohistochemistry assay was used to detect molecule expression in anterior cervical muscle tissues. Motor endplate activity was examined using the wholemount acetylcholinesterase staining method. The wet mass ratio of anterior cervical muscle was measured. The results indicated that Buyang Huanwu Tang treatment significantly alleviated inflammatory response, enhanced acetylcholinesterase activity, and motor endplate functions, and promoted wet mass of anterior cervical muscle compared to denervated-dependent skeletal muscle atrophy rat models ( < 0.05). Buyang Huanwu Tang regulated molecules of PI3K/PKB/GSK3β/FOXO1 signaling pathway. Buyang Huanwu Tang significantly reduced muscle atrophy F-box protein, MuFR-1, Bax and caspase 9 expression, significantly enhanced Bcl-2 expression, and remarkably increased element-binding protein and vascular endothelial growth factor levels, compared to Model group ( < 0.05). Buyang Huanwu Tang suppressed caspase 9 and caspase 3 activity and associated apoptosis. Moreover, PI3K specific blocker, LY294002, significantly inhibited the effects of Buyang Huanwu Tang on the above molecule expression ( < 0.05). In conclusion, Buyang Huanwu Tang improved motor endplate functions of denervated-dependent skeletal muscle atrophy rat model through suppressing mitochondria-mediated apoptosis and activating PI3K/PKB/FOXO1 signaling pathway.
失神经支配依赖性骨骼肌萎缩是一种由骨骼肌相关外周神经切断引起的疾病。其具体分子机制尚不清楚。采用中药补阳还五汤对失神经支配依赖性骨骼肌萎缩进行治疗，并应用于大鼠失神经支配依赖性骨骼肌萎缩模型，评估其在失神经支配依赖性骨骼肌萎缩发病机制中的治疗效果及相关机制。建立大鼠失神经支配依赖性骨骼肌萎缩模型，并随机分为八组，包括正常对照组、模型组、阳性对照组、模型+补阳还五汤组、模型+黄芪提取物组、模型+补阳还五汤-黄芪组、补阳还五汤+LY294002组和黄芪提取物+LY294002组。采用苏木精-伊红染色和定量逆转录-聚合酶链反应（qRT-PCR）检测肌肉组织的炎症反应。利用定量RT-PCR和蛋白质印迹法分析mRNA和蛋白质表达。采用免疫组织化学化学化学检测颈前肌组织中的分子表达。采用全层乙酰胆碱酯酶染色法检测运动终板活性。测量颈前肌的湿重比。结果表明，与失神经支配依赖性骨骼肌萎缩大鼠模型相比，补阳还五汤治疗显著减轻了炎症反应，增强了乙酰胆碱酯酶活性和运动终板功能，并促进了颈前肌的湿重（<0.05）。补阳还五汤调节PI3K/PKB/GSK3β/FOXO1信号通路分子。与模型组相比，补阳还五汤显著降低了肌肉萎缩F-box蛋白、MuFR-1、Bax和caspase 9的表达，显著增强了Bcl-2的表达，并显著提高了元件结合蛋白和血管内皮生长因子水平（<0.05）。补阳还五汤抑制caspase 9和caspase 3活性及相关凋亡。此外，PI3K特异性阻滞剂LY294002显著抑制补阳还五汤对上述分子表达的影响（<0.05）。总之，补阳还五汤通过抑制线粒体介导的凋亡和激活PI3K/PKB/FOXO1信号通路，改善了失神经支配依赖性骨骼肌萎缩大鼠模型的运动终板功能。
