颈椎病颈椎终板软骨的转录组学特征及针刀干预对FGF18/Akt轴关键分子表达的影响OA
Transcriptomic characteristics of cervical endplate cartilage in cervical spondylosis and effects of acupotomy on the FGF18/Akt axis with cervical spondylosis
目的 分析颈椎病(CS)终板软骨的转录组学特征及针刀对FGF18/Akt轴关键分子表达的影响.方法 基于GEO数据库结合生物信息学方法分析CS终板软骨的差异基因、关键信号通路等转录组学特征,并开展针刀干预CS兔模型的动物实验进行验证.24只SPF级新西兰兔随机分为正常组、CS组、针刀组,8只/组.除正常组兔外,余兔采用低头位法构建CS模型,采用X线验证CS模型是否成功构建.造模成功1周后,正常组和CS组兔正常饲养,针刀组兔进行针刀干预,1次/周,共干预3周.HE染色观察颈椎间盘病理结构改变,Tunel法检测细胞凋亡,实时荧光定量PCR检测椎间盘FGF18、FGFR3、Akt的mRNA表达,免疫组化法检测椎间盘髓核、终板软骨和纤维环的FGF18、p-Akt、Akt蛋白定位和表达情况.结果 与正常终板软骨相比,CS终板软骨存在大量差异表达基因,并显著富集于PI3K-Akt、钙离子、Rap1等信号通路;X线结果表明造模前后颈椎生理曲度、关节退行性改变等符合CS临床表现;HE染色结果发现CS颈椎间盘纹理杂乱,椎间盘纤维环变薄,髓核变小消失,针刀可缓解CS以上病理改变;qPCR结果发现与正常组兔相比,CS颈椎间盘FGF18、FGFR3、Akt表达差异无统计学意义(P>0.05),与CS组相比,针刀组FGF18、FGFR3、Akt表达差异亦无统计学意义(P>0.05);免疫组化结果表明,针刀可显著提高CS颈椎终板软骨FGF18及p-Akt的蛋白表达(P<0.05),降低CS颈椎终板软骨周围Akt与p-Akt的比值(P<0.05);Tunel显示CS组终板软骨凋亡细胞增多(P<0.05),针刀可减轻细胞凋亡(P<0.05).结论 CS颈椎终板软骨存在大量差异表达基因,针刀可能通过有效激活FGF18/Akt轴,减轻终板软骨细胞凋亡,从而延缓椎间盘退行性改变治疗CS.
Objective To explore the transcriptomic characteristics of the endplate cartilage of the cervical intervertebral disc in cervical spondylosis(CS)and the effects of acupotomy on expressions of fibroblast growth factor 18(FGF18)/protein kinase B(Akt)axis key molecules.Methods Transcriptomic analyses were performed using bioinformatics methods based on the GEO database.In the animal experiment,24 New Zealand rabbits were randomized equally into control,CS model,and acupotomy groups.In the latter two groups,CS models were established followed 7 days later by acupotomy intervention(once a week for 3 weeks)or no particular treatment.Histopathological changes and cell apoptosis in the intervertebral discs were examined with HE staining and TUNEL assay,and the mRNA expressions of FGF18,FGFR3,and Akt in the intervertebral discs were detected using RT-qPCR;the localization and expressions of FGF18,p-Akt,and Akt proteins were examined with immunohistochemistry.Results The endplate cartilage in CS exhibited numerous differentially expressed genes enriched in the PI3K-Akt signaling,calcium signaling,and Rap1 signaling pathways.CS rabbits showed obvious changes in the cervical spine curvature and joint degenerations,changes in cervical intervertebral disc texture,thinning of the annulus fibrosus,shrinkage or even absence of the nucleus pulposus,and increased apoptotic cells in the endplate cartilage,which were all obviously alleviated after acupotomy.No significant differences were found in the mRNA expressions of FGF18,FGFR3,and Akt in the cervical intervertebral discs among the 3 groups.Acupotomy significantly increased FGF18 and p-Akt protein expressions and reduced the Akt/p-Akt ratio in the cervical endplate cartilage of CS rabbits.Conclusion The cervical endplate cartilage of CS show numerous differentially expressed genes.Acupotomy may delay degenerative changes of the intervertebral discs and improves CS by activating the FGF18/Akt axis to reduce apoptosis of endplate cartilage cells.
刘福水;王小乐;KHALIUNAA Tumurbaatar;曹奇光;杨煜乾;任长安;朱金超;赵小兰;曹锂;邓彪
江西中医药大学附属医院骨伤九科,江西 南昌 330006||江西中医药大学临床医学院,江西 南昌 330006江西中医药大学附属医院骨伤九科,江西 南昌 330006江西中医药大学临床医学院,江西 南昌 330006江西中医药大学临床医学院,江西 南昌 330006江西中医药大学临床医学院,江西 南昌 330006江西中医药大学附属医院骨伤九科,江西 南昌 330006江西中医药大学临床医学院,江西 南昌 330006江西中医药大学临床医学院,江西 南昌 330006江西中医药大学临床医学院,江西 南昌 330006江西中医药大学临床医学院,江西 南昌 330006
颈椎病针刀转录组特征终板软骨FGF18/Akt轴
cervical spondylosisacupotomytranscriptome characteristicsintervertebral discFGF18/Akt axis
《南方医科大学学报》 2026 (1)
55-65,11
国家重点研发计划(2023YFC3502701)国家自然科学基金(82360940)江西省教育厅科技项目(GJJ2200990)江西省中医药管理局项目(2023B1191)Supported by National Key Research and Development Program of China(2023YFC3502701)National Natural Science Foundation of China(82360940).
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