Xie Yue, Zhang Jianpo, Yang Shengqi, Zhai Weifeng, Zhao Hailiang, Shen Zhan, Guo Ji, Jia Yongwei
Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China.
Guanghua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 200052, China.
Heliyon. 2024 Mar 19;10(6):e28095. doi: 10.1016/j.heliyon.2024.e28095. eCollection 2024 Mar 30.
To explore whether Bushen Huoxue Formula (BSHXF) improves the angiogenesis ability of transplanted endothelial progenitor cells (EPCs) in endplate and its potential mechanism in delaying intervertebral disc degeneration (IDD).
BSHXF was analyzed via Ultra-High Performance Liquid Chromatography (UPLC). Rabbit axial compression lumbar IDD models were constructed and the effects of BSHXF, EPCs, and their combination in IDD were determined by MRI, histological evaluation, TUNEL, and immunofluorescence assays. Additionally, CCK-8 assay, flow cytometry, and tube formation assay were used to evaluate EPCs viability, proliferation, cell cycle and the angiogenesis ability of EPCs between groups.
BSHXF and transplanted EPCs both attenuate the process of IDD in the rabbit model assessed by MRI, HE staining and Masson staining. TUNEL-positive NP cells were significantly reduced in the BSHXF group, EPCs group, and EPC + BSHXF group compared to the model group (P < 0.05), with the EPC + BSHXF group showing the most significant therapeutic effect. Immunofluorescence detection showed that VEGF, CD34 expression and quantity of microvessels in the endplate significantly increased in the EPC + BSHXF group compared to all the other groups (P < 0.05). Besides, the CCK-8 assay showed an upregulation of EPC viability and the tube formation assay demonstrated a significant increase in tube length and branching in EPCs cultured with BSHXF-containing serum (P < 0.05). Furthermore, BSHXF-containing serum increased VEGF expression in EPCs cultured in vitro (P < 0.05).
Both BSHXF and EPCs transplantation play an important role in increasing endplate angiogenesis and attenuating IDD. BSHXF can enhance the viability and tube-forming ability of EPCs and endplate microcirculation.
探讨补肾活血方（BSHXF）是否能提高移植的内皮祖细胞（EPCs）在终板的血管生成能力及其延缓椎间盘退变（IDD）的潜在机制。
采用超高效液相色谱（UPLC）分析补肾活血方。构建兔轴向压缩腰椎间盘退变模型，通过磁共振成像（MRI）、组织学评估、TUNEL和免疫荧光分析确定补肾活血方、内皮祖细胞及其联合应用对椎间盘退变的影响。此外，采用CCK-8法、流式细胞术和管腔形成试验评估各组内皮祖细胞的活力、增殖、细胞周期及血管生成能力。
通过MRI、苏木精-伊红（HE）染色和马松染色评估，补肾活血方和移植的内皮祖细胞均能减轻兔模型中的椎间盘退变过程。与模型组相比，补肾活血方组、内皮祖细胞组和内皮祖细胞+补肾活血方组的TUNEL阳性髓核细胞显著减少（P<0.05），其中内皮祖细胞+补肾活血方组的治疗效果最为显著。免疫荧光检测显示，与其他所有组相比，内皮祖细胞+补肾活血方组终板中血管内皮生长因子（VEGF）、CD34表达及微血管数量显著增加（P<0.05）。此外，CCK-8法显示内皮祖细胞活力上调，管腔形成试验表明，用含补肾活血方血清培养的内皮祖细胞的管腔长度和分支显著增加（P<0.05）。此外，含补肾活血方血清增加了体外培养的内皮祖细胞中VEGF的表达（P<0.05）。
补肾活血方和内皮祖细胞移植在增加终板血管生成和减轻椎间盘退变方面均起重要作用。补肾活血方能增强内皮祖细胞的活力和管腔形成能力以及终板微循环。
