Chen Guiling, Liu Aoyin, He Meixing, Zheng Xianhui, Zhu Wei
The Second Clinical College of Guangzhou University of Chinese Medicine, Guangzhou 510120, China.
The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510120, China.
ACS Omega. 2025 Jul 20;10(29):31638-31648. doi: 10.1021/acsomega.5c02499. eCollection 2025 Jul 29.
This study aims to investigate the effects of TGL (total glycosides from lily) and its constituent regaloside B, when absorbed in the blood, on the osteogenic differentiation, migration, and angiogenesis of human umbilical cord-derived mesenchymal stem cells (hUCMSCs).
hUCMSCs were treated with TNF-α and IFN-γ to induce an inflammatory microenvironment mimicking conditions. Cell cycle progression and surface markers of the hUCMSCs were analyzed by flow cytometry. Alizarin Red staining was performed to quantify extracellular calcium deposition, reflecting the osteogenic effects of TGL and regaloside B. The relative mRNA and protein expression levels of PTGS1, VEGF, HIF-1α, and CCL5 were assessed by qRT-PCR and Western blotting (WB). Cell migration ability was examined using wound healing assays. TGL components in rat plasma were identified by UHPLC-LTQ-Orbitrap-MS analysis. The effects of regaloside B on CCL5, CXCL9, CXCL10, VCAM, ICAM, IL8, and IDO mRNA levels were verified under the induction of TNF-α and IFN-γ.
TGL suppressed TNF-α/IFN-γ-induced expression of chemokine CCL5 and angiogenic factors (PTGS1, VEGF), inhibited cell migration, and enhanced osteogenic differentiation of hUCMSCs ( < 0.05). Pharmacokinetic analysis revealed that regaloside B, a systemic metabolite of TGL detected in rat plasma, significantly inhibited TNF-α and IFN-γ-induced chemokines and angiogenic factors in hUCMSCs in comparison with the same dose of TGL ( < 0.05).
TGL and its bioactive metabolite regaloside B demonstrated modulatory effects on TNF-α/IFN-γ-induced alterations in osteogenic differentiation, migratory capacity, and angiogenesis regulatory potential of hUCMSCs, suggesting that regaloside B mediates the therapeutic effects of TGL.
本研究旨在探讨百合总苷（TGL）及其成分瑞香苷B吸收入血后对人脐带间充质干细胞（hUCMSCs）成骨分化、迁移及血管生成的影响。
用肿瘤坏死因子-α（TNF-α）和干扰素-γ（IFN-γ）处理hUCMSCs以诱导模拟炎症微环境。通过流式细胞术分析hUCMSCs的细胞周期进程和表面标志物。进行茜素红染色以定量细胞外钙沉积，反映TGL和瑞香苷B的成骨作用。通过实时定量聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法（WB）评估环氧化酶-1（PTGS1）、血管内皮生长因子（VEGF）、缺氧诱导因子-1α（HIF-1α）和趋化因子配体5（CCL5）的相对mRNA和蛋白质表达水平。使用伤口愈合试验检测细胞迁移能力。通过超高效液相色谱-线性离子阱-轨道阱质谱（UHPLC-LTQ-Orbitrap-MS）分析鉴定大鼠血浆中的TGL成分。在TNF-α和IFN-γ诱导下验证瑞香苷B对CCL5、CXC趋化因子配体9（CXCL9）、CXC趋化因子配体10（CXCL10）、血管细胞黏附分子（VCAM）、细胞间黏附分子（ICAM）、白细胞介素8（IL8）和吲哚胺2,3-双加氧酶（IDO）mRNA水平的影响。
TGL抑制TNF-α/IFN-γ诱导的趋化因子CCL5和血管生成因子（PTGS1、VEGF）的表达，抑制细胞迁移，并增强hUCMSCs的成骨分化（<0.05）。药代动力学分析显示，瑞香苷B是在大鼠血浆中检测到的TGL的全身代谢产物，与相同剂量的TGL相比，其显著抑制TNF-α和IFN-γ诱导的hUCMSCs中的趋化因子和血管生成因子（<0.05）。
TGL及其生物活性代谢产物瑞香苷B对TNF-α/IFN-γ诱导的hUCMSCs成骨分化、迁移能力和血管生成调节潜能的改变具有调节作用，提示瑞香苷B介导了TGL的治疗作用。
