电针对颈椎病家兔髓核细胞自噬相关蛋白表达的影响OA北大核心CSTPCDMEDLINE
Effect of electroacupuncture on expression of autophagy-related proteins in nucleus pulposus of cervical spondylosis rabbits
目的:观察电针对颈椎病模型家兔椎间盘组织形态,髓核细胞凋亡情况及髓核组织自噬启动蛋白1(ULK1)、酵母自噬相关基因6同源物(Beclin1)和微管相关蛋白轻链3(LC3)表达的影响,从细胞自噬途径探讨电针治疗颈椎病的作用机制.方法:24只新西兰家兔随机分为空白组、模型组、电针组,每组各8只.电针组取双侧颈(C)3~C6"夹脊"行电针刺激,25 min/次,1次/d,5次为1个疗程,疗程间隔2d,共干预4个疗程.采用X线检查颈椎影像变化并评价影像学评分;透射电镜观察髓核细胞超微结构变化;HE染色观察椎间盘组织形态变化并进行病理评分;TUNEL染色观察髓核细胞凋亡率;Western blot法检测髓核组织ULK1、Beclin1、LC3蛋白的表达水平.结果:与空白组比较,模型组家兔颈椎影像学评分明显升高(P<0.01),椎间盘组织病理评分明显升高(P<0.05),髓核细胞凋亡率明显升高(P<0.01),髓核组织ULK1、Beclin1、LC3Ⅱ蛋白表达水平降低(P<0.05).与模型组比较,电针组椎间盘病理评分明显降低(P<0.05),髓核细胞凋亡率降低(P<0.01),髓核组织ULK1、Beclin1、LC3 Ⅱ蛋白表达水平升高(P<0.01).空白组家兔颈椎间盘髓核组织细胞器结构基本正常,并出现少量环状隔离膜结构,为自噬的初期阶段;模型组椎间盘髓核组织细胞器结构遭到破坏,出现胞质浓缩现象;电针组髓核细胞出现具有双层膜结构的自噬小体.结论:电针能够促进髓核组织ULK1、Beclin1、LC3Ⅱ蛋白的表达,减少髓核细胞凋亡,改善椎间盘退变.
Objective To observe the effects of electroacupuncture(EA)on the morphological changes of intervertebral disc tissues,apoptosis of nucleus pulposus cells,and the protein expression of Unc-51 like autophagy-activated kinase 1(ULK1),homologous series of yeast Atg6(Beclin 1),and light chain protease complication 3 type(LC3)in nucleus pulposus tissue of cervical spondylosis rabbits,so as to explore the role of cellular autophagy in EA treatment of cervical spondylosis.Methods A total of 24 New Zealand white rabbits were randomly divided into blank,model and EA groups,with 8 rabbits in each group.In the EA group,both sides of the cervical(C)3-C6"Jiaji"(EX-B2)were stimulated by EA(2 Hz/100 Hz,1 mA)for 25 min,once daily for 5 days in a course,with a 2-day interval between courses,totaling 4 treatment courses.X-ray was used to assess cervical spine radiographic changes and evaluate radiographic scores;transmission electron microscopy was used to observe ultrastructural changes in nucleus pulposus cells;HE staining was used to observe morphological changes of intervertebral disc tissues and conduct pathological scoring;TUNEL staining was used to observe apoptosis rate of nucleus pulposus cells;Western blot was performed to detect protein expression levels of ULK1,Beclin1,and LC3 in nucleus pulposus tissue.Results Compared with the blank group,rabbits in the model group showed significantly higher cervical spine radiographic scores(P<0.01),higher pathological scores of intervertebral disc tissues(P<0.05),increased apoptosis rate of nucleus pulposus cells(P<0.01),and decreased expression levels of ULK1,Beclin1,and LC3 Ⅱ proteins in nucleus pulposus tissue(P<0.05).Compared with the model group,the EA group showed significantly lower pathological scores of intervertebral discs(P<0.05),lower apoptosis rate of nucleus pulposus cells(P<0.01),and higher protein expression levels of ULK1,Beclin1,and LC3 Ⅱ in nucleus pulposus tissue(P<0.01).Rabbits in the blank control group exhibited generally normal organelle structures in nucleus pulposus tissues with few autophagic vacuoles,indicative of early stages of autophagy;while those in the model group showed disrupted organelle structures with cytoplasmic condensation and those in the EA group exhibited autophagosomes with double-membrane structures in nucleus pulposus tissues.Conclusion EA promotes the expression of ULK1,Beclin1,and LC3 Ⅱ proteins in nucleus pulposus tissues,reduces apoptosis of nucleus pulposus cells,and improves intervertebral disc degeneration.
陈佳丽;赵子莹;高航;徐银琴;王光义
贵州医科大学临床医学院,贵阳 550004贵州医科大学附属医院针灸科,贵阳 550004
电针颈椎病椎间盘退变髓核自噬
ElectroacupunctureCervical spondylosisIntervertebral disc degenerationNucleus pulposusAutophagy
《针刺研究》 2024 (008)
821-828 / 8
贵阳市科技计划项目(No.[2019]9-1-4);吕明庄全国名老中医药专家传承工作室织金县工作站项目
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