Xin Li, Feng Huan-Cun, Zhang Qiang, Cen Xiao-Lin, Huang Rong-Rong, Tan Guo-Yao, Zhang Qun
Office of Clinical Trial of Drug, Guangdong Provincial Key Laboratory of Bone and Joint Degeneration Diseases, The Third Affiliated Hospital, Southern Medical University, Guangzhou, 510630, Guangdong, China; Department of Pharmacy, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, 510120, Guangdong, China.
Office of Clinical Trial of Drug, Guangdong Provincial Key Laboratory of Bone and Joint Degeneration Diseases, The Third Affiliated Hospital, Southern Medical University, Guangzhou, 510630, Guangdong, China.
J Ethnopharmacol. 2025 Feb 10;338(Pt 1):119023. doi: 10.1016/j.jep.2024.119023. Epub 2024 Nov 1.
Osteoporosis (OP) is a degenerative bone disease commonly associated with reduced bone density and increased fracture risk.
This study aimed to validate the therapeutic effects of Simiao wan (SMW) on OP and explore the underlying mechanism, particularly focusing on the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway.
The chemical components of SMW were identified using UPLC-Q-TOF-MS/MS. The obtained compounds were then input into the TCMSP, TargetNet, and SwissTargetPrediction databases to predict potential targets. OP-related targets were collected from the GeneCards and DisGeNET databases, and intersecting targets were identified through a Venn diagram. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed on the intersecting targets using the Database for Annotation, Visualization and Integrated Discovery (DAVID). SMW extract was subsequently used to treat osteoblasts in vitro, and its toxicity on osteoblasts was assessed using Cell Counting Kit-8 (CCK-8) and lactate dehydrogenase (LDH) release assays. Osteoblast differentiation and activity were further evaluated using alizarin red staining, alkaline phosphatase staining, and Western blot analyses to validate the activation of network pharmacological signaling pathways.
A total of 121 potential targets were identified for SMW in the treatment of OP, with AKT1 as the primary target. The PI3K/AKT pathway emerged as a key signaling pathway potentially involved in SMW's therapeutic effects o OP. Toxicity assessments showed no significant toxicity of SMW on osteoblasts. Additionally, SMW promoted osteoblast proliferation, alkaline phosphatase activity, calcium nodule deposition, and the expression of osteogenic markers (osteocalcin (OCN), runt-related transcription factor 2 (RunX2), and collagen I), and activated the PI3K/AKT signaling pathway. The PI3K/AKT pathway inhibitor LY294002 partially reversed the SMW-induced mineral deposition and expression of OCN, RunX2, and collagen I.
SMW demonstrated effective multi-target and multi-pathway therapeutic potential in the treatment of OP, with a significant impact on the PI3K/AKT signaling pathway.
骨质疏松症（OP）是一种退行性骨病，通常与骨密度降低和骨折风险增加有关。
本研究旨在验证四妙丸（SMW）对OP的治疗效果，并探索其潜在机制，尤其关注磷脂酰肌醇3激酶（PI3K）/蛋白激酶B（AKT）信号通路。
采用超高效液相色谱-四极杆飞行时间串联质谱（UPLC-Q-TOF-MS/MS）鉴定SMW的化学成分。将所得化合物输入中药系统药理学数据库与分析平台（TCMSP）、TargetNet和瑞士药物靶点预测数据库（SwissTargetPrediction）以预测潜在靶点。从基因卡片（GeneCards）和疾病基因数据库（DisGeNET）收集OP相关靶点，并通过韦恩图确定交集靶点。使用注释、可视化和综合发现数据库（DAVID）对交集靶点进行基因本体论（GO）和京都基因与基因组百科全书（KEGG）富集分析。随后用SMW提取物体外处理成骨细胞，并用细胞计数试剂盒-8（CCK-8）和乳酸脱氢酶（LDH）释放试验评估其对成骨细胞的毒性。使用茜素红染色、碱性磷酸酶染色和蛋白质印迹分析进一步评估成骨细胞分化和活性，以验证网络药理学信号通路的激活。
共鉴定出121个SMW治疗OP的潜在靶点，其中AKT1为主要靶点。PI3K/AKT通路是可能参与SMW对OP治疗作用的关键信号通路。毒性评估显示SMW对成骨细胞无明显毒性。此外，SMW促进成骨细胞增殖、碱性磷酸酶活性、钙结节沉积以及成骨标志物（骨钙素（OCN）、 runt相关转录因子2（RunX2）和I型胶原）的表达，并激活PI3K/AKT信号通路。PI3K/AKT通路抑制剂LY294002部分逆转了SMW诱导的矿物质沉积以及OCN、RunX2和I型胶原的表达。
SMW在治疗OP方面显示出有效的多靶点、多途径治疗潜力，对PI3K/AKT信号通路有显著影响。
