College of Basic Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai, China.
Experiment Center for Science and Technology, Shanghai University of Traditional Chinese Medicine, Shanghai, China.
J Ethnopharmacol. 2022 Aug 10;294:115323. doi: 10.1016/j.jep.2022.115323. Epub 2022 Apr 26.
In traditional Chinese medicine (TCM), Zuogui Wan (ZGW) is a classical prescription for senile disorders and delay aging. Modern studies show that ZGW promotes central nerve cell regeneration, prevents and cures osteoporosis, enhances the body's antioxidant capacity, regulates the body's immune function, and promotes mesenchymal stem cells (MSCs) proliferation.
It has been shown that MSCs aging is closely associated with organism's aging and age-related disorders. The study aimed to define the effects of ZGW on the aging bone marrow mesenchymal stem cells (BMSCs) and to identify the mechanisms of ZGW delaying BMSCs senescence.
Network pharmacology analysis combined with GEO data mining, molecular docking and experimental validation were used to evaluate the mechanisms by which ZGW delays MSCs senescence (MSCS). LC-MS was used for quality control analysis of ZGW.
PPI network analysis revealed that EGF, TNF, JUN, MMPs, IL-6, MAPK8, and MYC are components of the core PPI network. GO and KEGG analyses revealed that oxidative stress, regulation of response to DNA damage stimuli, and Wnt signaling were significantly enriched. GEO database validation also indicated that Wnt signaling closely correlated with MSCs aging. Molecular docking analysis of the top-13 active components in the "ZGW-Targets-MSCS" network indicated that most components have strong affinity for key proteins in Wnt signaling, suggesting that modulation of Wnt signaling is an important mechanism of ZGW activity against MSCS. Further experimental validation found that ZGW indeed regulates Wnt signaling and suppresses the expression of age-related factors to enhance cell proliferation, ameliorate DNA damage, and reduce senescence-related secretory phenotype (SASP) secretion, thereby maintaining multidirectional differentiation of rat BMSCs. Similar results were obtained using the Wnt inhibitor, XAV-939.
Together, our data show that ZGW slows BMSCs aging by suppressing Wnt signaling.
在传统中医（TCM）中，左归丸（ZGW）是一种治疗老年疾病和延缓衰老的经典方剂。现代研究表明，ZGW 能促进中枢神经细胞再生，预防和治疗骨质疏松症，增强机体抗氧化能力，调节机体免疫功能，促进间充质干细胞（MSCs）增殖。
已证明 MSCs 的衰老与机体的衰老和与年龄相关的疾病密切相关。本研究旨在确定 ZGW 对衰老骨髓间充质干细胞（BMSCs）的作用，并确定 ZGW 延缓 BMSCs 衰老的机制。
采用网络药理学分析结合 GEO 数据挖掘、分子对接和实验验证的方法，评价 ZGW 延缓 MSCs 衰老的机制（MSCS）。LC-MS 用于 ZGW 的质量控制分析。
PPI 网络分析显示，EGF、TNF、JUN、MMPs、IL-6、MAPK8 和 MYC 是核心 PPI 网络的组成部分。GO 和 KEGG 分析显示，氧化应激、对 DNA 损伤刺激的反应调节以及 Wnt 信号通路显著富集。GEO 数据库验证也表明，Wnt 信号通路与 MSCs 衰老密切相关。“ZGW-Targets-MSCS”网络中前 13 种活性成分的分子对接分析表明，大多数成分与 Wnt 信号通路中的关键蛋白具有很强的亲和力，提示调节 Wnt 信号通路是 ZGW 活性对抗 MSCs 的重要机制。进一步的实验验证发现，ZGW 确实调节了 Wnt 信号通路，抑制了与年龄相关的因子的表达，从而增强了细胞的增殖能力，改善了 DNA 损伤，减少了衰老相关的分泌表型（SASP）分泌，从而维持了大鼠 BMSCs 的多向分化。使用 Wnt 抑制剂 XAV-939 也得到了类似的结果。
综上所述，我们的数据表明，ZGW 通过抑制 Wnt 信号通路来减缓 BMSCs 的衰老。
