数字中医药（英文）2026，Vol.9Issue(3)：397-411,15.DOI:10.1016/j.dcmed.2026.08.007
补肾活血方通过调控内皮祖细胞SIRT1/CXCL12/CXCR4/CXCR7轴对大鼠椎间盘退变及终板微循环的保护作用
Protective effects of Bushen Huoxue Formula on intervertebral disc degeneration and endplate microcirculation in rats via the SIRT1/CXCL12/CXCR4/CXCR7 axis in endothelial progenitor cells
摘要
Abstract
Objective To investigate the efficacy of the Bushen Huoxue Formula(补肾活血方,BSHXF)in alleviating intervertebral disc degeneration(IVDD),improving endplate microcirculation,and enhancing endothelial progenitor cell(EPC)function,while investigating the potential role of the sirtuin 1(SIRT1)/C-X-C motif chemokine ligand 12(CXCL12)/C-X-C motif chemokine receptor 4(CXCR4)/C-X-C motif chemokine receptor 7(CXCR7)signaling path-way. Methods This study comprised both in vivo and in vitro experiments.In vivo,24 male Sprague-Dawley(SD)rats(4-6 weeks old)were used,and allocated into four groups(n=6 per group):Rat-Control,Rat-Model,Rat-BSHXF,and Rat-EX527+BSHXF groups.IVDD was induced by applying continuous axial compression(approximately 11 N)to the caudal inter-vertebral discs for two weeks.Following model establishment,rats in the Rat-BSHXF group received oral BSHXF at 17.71 g/(kg·d)once daily,whereas those in the Rat-EX527+BSHXF group received the SIRT1 inhibitor EX527 at 5 mg/kg by intraperitoneal injection once daily in addition to the same BSHXF regimen for two consecutive weeks.Disc degeneration was eval-uated using magnetic resonance imaging(MRI),Pfirrmann grading,and hematoxylin and eosin(HE)staining.Endplate microvascular density was assessed via CD31 immunostaining.The mRNA expression levels of SIRT1,CXCL12,CXCR4,and CXCR7 in disc and endplate tis-sues were quantified using reverse transcription quantitative polymerase chain reaction(RT-qPCR),whereas their protein expression levels were assessed using Western blot and im-munofluorescence staining.In vitro,rat bone marrow-derived EPCs were isolated from a sep-arate cohort of six SD rats,cultured,and identified via CD34/CD133 immunofluorescence staining.The EPCs were allocated into Control,BSHXF,si-SIRT1,si-SIRT1+BSHXF,and Celecoxib groups.Cells in the BSHXF-related groups were treated with 10%BSHXF-contain-ing serum prepared using another separate cohort of six SD rats,and SIRT1 knockdown was achieved using SIRT1 targeting small interfering RNA(si-SIRT1).EPC viability,migration,and tube formation were evaluated using cell counting kit-8(CCK-8),transwell migration,and Matrigel tube formation assays,respectively.Additionally,the mRNA and protein expression levels of SIRT1,CXCL12,CXCR4,and CXCR7 in EPCs were measured via RT-qPCR and West-ern blot,respectively. Results In vivo,axial compression induced significant disc degeneration,characterized by reduced T2 signal intensity,decreased disc height,and Pfirrmann grade IV-V changes.Compared with the Rat-Model and Rat-EX527+BSHXF groups,the Rat-BSHXF group exhibit-ed less severe histological degeneration and significantly increased endplate microvascular density(P<0.05).Furthermore,BSHXF significantly upregulated the mRNA and protein ex-pression levels of SIRT1,CXCL12,CXCR4,and CXCR7 in disc and endplate tissues compared with both the Rat-Model and Rat-EX527+BSHXF groups(P<0.05).These elevated expres-sion levels in the Rat-BSHXF group were further corroborated by immunofluorescence analy-sis(P<0.05).In vitro,treatment with BSHXF-containing serum significantly enhanced EPC viability,migration,total tube length,and branch point formation compared with the Control,si-SIRT1,and si-SIRT1+BSHXF groups(P<0.05).No significant differences in tube forma-tion were observed between the BSHXF and Celecoxib groups(P>0.05).Conversely,SIRT1 knockdown markedly impaired EPCs angiogenesis-related functions and abrogated the BSHXF-induced enhancements in EPCs viability,migration,and tube formation,with no sig-nificant differences observed between the si-SIRT1 and si-SIRT1+BSHXF groups(P>0.05). Conclusion BSHXF attenuated IVDD-related degenerative changes and increased endplate microvascular density in a rat model of caudal disc compression,while enhancing EPC viabil-ity,migration,and angiogenesis in vitro.These findings suggest that the effects of BSHXF on endplate microcirculation during IVDD may be associated with the SIRT1-related modula-tion of the CXCL12/CXCR4/CXCR7 axis in EPCs.关键词
椎间盘退变/补肾活血方/终板微循环/内皮祖细胞/SIRT1/CXCL12/CXCR4/CXCR7轴Key words
Intervertebral disc degeneration/Bushen Huoxue Formula/Endplate microcirculation/Endothelial progenitor cells/SIRT1/CXCL12/CXCR4/CXCR7 axis引用本文复制引用
杨升祺,申展,翟伟峰,谢越,贾永伟,郭际..补肾活血方通过调控内皮祖细胞SIRT1/CXCL12/CXCR4/CXCR7轴对大鼠椎间盘退变及终板微循环的保护作用[J].数字中医药（英文）,2026,9(3):397-411,15.基金项目
National Natural Science Foundation of China(82575112 and 82304955),and Shanghai Pudong New Area Science and Technology Development Fund,Public Institution Livelihood Research Special Project(PKJ2024-Y47). （82575112 and 82304955）
