中国组织工程研究

中国组织工程研究 ›› 2025, Vol. 29 ›› Issue (2): 331-338.doi: 10.12307/2025.204

• 脊柱组织构建 spinal tissue construction • 上一篇    下一篇

超声引导下针刀松解黄韧带对兔退变腰椎间盘整合素α5、β1表达的影响

陈  灿1，赵  宇1，胡斌涵 1，杜梦凡1，刘俊宁 1，2，牛素生1，2，张  燕1，2   

  1. 福建中医药大学，1中医学院，2中医骨伤及运动康复教育部重点实验室，福建省福州市  350122
  • 收稿日期:2023-12-05 接受日期:2024-01-20 出版日期:2025-01-18 发布日期:2024-05-25
  • 通讯作者: 张燕，副教授，福建中医药大学，中医学院，中医骨伤及运动康复教育部重点实验室，福建省福州市 350122
  • 作者简介:陈灿，女，1999年生，江苏省淮安市人，汉族，在读硕士，主要从事中医药防治骨关节疾病的基础与临床研究。
  • 基金资助:
    福建省自然科学基金项目(2023J01862)，项目负责人：张燕；福建中医药大学基础类学科科研提升计划项目(XJC2022008)，项目负责人：张燕

Effect of ultrasound-guided needle-knife release of the ligamentum flavum on the expression of integrin alpha5 and beta1 in degenerative rabbit lumbar intervertebral discs

Chen Can1, Zhao Yu1, Hu Binhan1, Du Mengfan1, Liu Junning1, 2, Niu Susheng1, 2, Zhang Yan1, 2   

  1. 1School of Traditional Chinese Medicine, Fuiian University of Traditional Chinese Medicine, Fuzhou 350122, Fujian Province, China; 2Key Laboratory of Orthopedics & Traumatology of Traditional Chinese Medicine and Rehabilitation, Ministry of Education, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, Fujian Province, China
  • Received:2023-12-05 Accepted:2024-01-20 Online:2025-01-18 Published:2024-05-25
  • Contact: Zhang Yan, Associate professor, School of Traditional Chinese Medicine, Fuiian University of Traditional Chinese Medicine, Fuzhou 350122, Fujian Province, China; Key Laboratory of Orthopedics & Traumatology of Traditional Chinese Medicine and Rehabilitation, Ministry of Education, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, Fujian Province, China
  • About author:Chen Can, Master candidate, School of Traditional Chinese Medicine, Fuiian University of Traditional Chinese Medicine, Fuzhou 350122, Fujian Province, China
  • Supported by:
    the Natural Science Foundation of Fujian Province, NO. 2023J01862 (to ZY); Basic Discipline Research Enhancement Program of Fujian University of Traditional Chinese Medicine, No. XJC2022008 (to ZY)

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摘要：

文题释义：
超声引导：随着超声技术的发展，介入超声技术被广泛用于临床，实时可视图像引导显著提高了干预治疗措施的准确性和安全性，可以有效避免误伤神经、血管等重要组织。
整合素：是一种存在于细胞表面的力学感受器和黏附分子，能维持细胞结构和力学完整性、感受机械应力刺激，具有促进力学信号转变为生物学信号的功能，其中整合素α5、β1是纤维连接蛋白的特异性受体。

背景：针刀松解黄韧带可有效改善腰椎退行性患者的症状，超声引导可增加针刀松解的精准性，但针刀松解黄韧带后对退变椎间盘的具体影响及可能的机制尚待明确。
目的：探讨超声引导下针刀松解黄韧带对兔腰椎间盘退变的作用。
方法：取24只新西兰兔，采用随机数字表法分为对照组(n=6)与造模组(n=18)。造模组采用横断L5/6、L6/7节段棘上、棘间韧带并保持站立姿势对腰椎施加轴向载荷的方式建立兔腰椎间盘退变模型，造模成功后再随机分为模型组、超声针刀组、假针刀组，每组6只，超声针刀组在超声引导下使用针刀松解L7/S1右侧黄韧带，假针刀组针刀入路同超声针刀组，但不松解黄韧带，1次/周，共4次。针刀干预后30 d，利用MRI观察L7/S1椎间盘髓核信号强度变化，苏木精-伊红染色观察L7/S1椎间盘形态变化，免疫组化染色检测L7/S1椎间盘髓核内Ⅰ，Ⅱ型胶原表达，RT-PCR、Western Blot检测L7/S1椎间盘中整合素α5、β1、p38、核因子κB的表达。
结果与结论：①MRI成像显示，模型组兔椎间盘髓核呈灰-黑色，髓核灰度值明显低于对照组(P < 0.01)；超声针刀组兔椎间盘髓核亮度较模型组升高，髓核灰度值高于模型组(P < 0.01)；②苏木精-伊红染色显示，模型组髓核形状不规则，髓核细胞数量减少，细胞外基质被压缩，纤维环破裂，终板结构与边界不清，软骨细胞排列紊乱；与模型组相比，超声针刀组髓核细胞数量增多，纤维环破裂情况有改善，软骨终板细胞排列较为规则；③免疫组化染色显示，与对照组比较，模型组髓核内Ⅰ型胶原阳性表达增加(P < 0.01)，Ⅱ型胶原阳性表达减少(P < 0.01)；与模型组比较，超声针刀组髓核内Ⅰ型胶原阳性表达减少(P < 0.01)，Ⅱ型胶原阳性表达增加(P < 0.01)；④RT-PCR、Western Blot检测显示，与对照组比较，模型组兔椎间盘中整合素α5、整合素β1、p38、核因子κB的mRNA与蛋白表达均升高(P < 0.01)；与模型组比较，超声针刀组兔椎间盘中整合素α5、整合素β1、p38、核因子κB的mRNA与蛋白表达均降低(P < 0.01)；⑤结果表明，超声引导下针刀松解黄韧带可改善兔腰椎间盘退变程度，该作用与通过调控整合素α5、β1表达、抑制p38和核因子κB表达有关。

https://orcid.org/0009-0002-5976-2985(陈灿)
中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 腰椎间盘退变, 针刀, 黄韧带, 超声引导, 整合素

Abstract: BACKGROUND: Needle-knife release of the ligamentum flavum can effectively improve symptoms in patients with lumbar degeneration, and ultrasound guidance can increase the precision of needle-knife release; however, the specific effects of needle-knife release of the ligamentum flavum on the degenerated intervertebral discs and the possible mechanisms remain to be clarified.
OBJECTIVE: To investigate the effect of ultrasound-guided needle-knife release of the ligamentum flavum. 
METHODS: Twenty-four New Zealand rabbits were randomized into control (n=6) and model (n=18) groups. A rabbit model of lumbar disc degeneration model was established in the model group by cutting the supraspinous and interspinous ligaments of the L5/6 and L6/7 segments to maintain a standing posture and apply axial load to the lumbar spine. After successful modeling, the model rabbits were subdivided into a control group, a model group, an ultrasonic needle-knife group, and a sham needle-knife group according to a random number table method, with six animals in each group. The ultrasonic needle-knife group underwent ultrasound-guided needle-knife release of the right yellow ligament of L7/S1, once every week, for a total of four times. The needle-knife approach in the sham needle-knife group was the same as that in the ultrasound needle-knife group, but the ligamentum flavum was not released. At 30 days after the intervention, MRI was used to observe the changes in the signal intensity of the nucleus pulposus within the L7/S1 segment. Hematoxylin-eosin staining was used to observe the morphological changes of the L7/S1 segment. Immunohistochemical staining was used to detect the expression of type I and II collagen in the nucleus pulposus of the L7/S1 segment. RT-PCR and western blot were used to detect the expression of integrin α5 and β1, p38, and nuclear factor κB in the L7/S1 segment.
RESULTS AND CONCLUSION: MRI findings indicated that the nucleus pulposus of the intervertebral disc of rabbits in the model group was gray-black in color, and the gray value of the nucleus pulposus was significantly lower than that of the control group (P < 0.01). The brightness of the nucleus pulposus of the intervertebral disc of the rabbits in the ultrasonic needle-knife group was elevated compared with that of the model group, and the gray value of the nucleus pulposus was higher than that of the model group (P < 0.01). Results from hematoxylin-eosin staining showed that in the model group, the shape of the nucleus pulposus was irregular, the number of nucleus pulposus cells was reduced, the extracellular matrix was compressed, the fibrous ring was ruptured, the structure and boundary of the end plate were unclear, and the chondrocytes were arranged disorderly. Compared with the model group, the ultrasonic needle-knife group showed an increase in the number of the nucleus pulposus, an improvement in the rupture of the fibrous ring, and more regular arrangement of cartilage endplate cells. Results from immunohistochemical staining showed an increase in positive expression of type I collagen (P < 0.01) and a decrease in positive expression of type II collagen in the nucleus pulposus of the model group compared with the control group as well as a decrease in positive expression of type I collagen and an increase in positive expression of type II collagen in the nucleus pulposus of the ultrasonic needle-knife group compared with the model group (P < 0.01). RT-PCR and western blot assays showed that the mRNA and protein expression of integrin α5, integrin β1, p38, and nuclear factor κB in the intervertebral discs of rabbits in the model group were increased compared with that in the control group (P < 0.01); the mRNA and protein expression of integrin α5, integrin β1, p38, and nuclear factor κB in the intervertebral discs of rabbits in the ultrasonic needle-knife group was decreased compared with that in the model group (P < 0.01). To conclude, ultrasound-guided needle-knife release of the ligamentum flavum can improve the degree of lumbar disc degeneration in rabbits, which may be related to the inhibition of p38 and nuclear factor-κB expression by modulating integrin α5 and β1 expression.

Key words: lumbar disc degeneration, needle-knife, ligamentum flavum, ultrasound guidance, integrin

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