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[电针“夹脊”(EX - B 2)对软骨终板Modic改变家兔软骨细胞外基质及炎症反应的影响]

[Effects of electroacupuncture at "Jiaji" (EX-B 2) on extracellular matrix of chondrocytes and inflammatory reaction in rabbits with Modic changes of cartilage endplate].

作者信息

Wang Min, Zou Jing, Huang Guofu

机构信息

College of Acupuncture-Moxibustion and Orthopedics, Hubei University of CM, Wuhan 430065, China.

Deparment of Acupuncture and Moxibustion, Wuhan Hospital of Tradition Chinese and Western Medicine.

出版信息

Zhongguo Zhen Jiu. 2024 Sep 12;44(9):1046-53. doi: 10.13703/j.0255-2930.20230928-k0002.

Abstract

OBJECTIVE

To observe the effects of electroacupuncture (EA) at "Jiaji" (EX-B 2) on extracellular matrix (ECM) of chondrocytes and inflammatory reaction in rabbits with Modic changes (MC) of cartilage endplate, and to explore the mechanism of EA in treating MC of endplate cartilage.

METHODS

Eighteen male New Zealand white rabbits were randomly divided into a sham operation group, a model group and an EA group, 6 rabbits in each group. Based on the autoimmune theory, MC model was established by embedding autogenous nucleus pulposus in the rabbits of the model group and the EA group, based on autoimmunity. After successful modeling, EA was applied at bilateral "Jiaji" (EX-B 2) of L and L in the EA group, with disperse-dense wave, 2 Hz/15 Hz in frequency and 1 mA in current intensity, 20 min a time, once a day, 1-day interval was taken after continuous 6-day intervention, for 4 weeks totally. Before and after modeling, as well as before and after intervention, the comprehensive response score was observed. After modeling and intervention, magnetic resonance imaging (MRI) was used to observe the signal intensity of intervertebral disc and cartilage endplate. After intervention, the morphology of chondrocytes of cartilage endplate was observed by HE staining; the positive expression of a disintegrin and metalloproteinase with thrombospondin motif-5 (ADAMTS5) and Aggrecan in the cartilage endplate was detected by immunohistochemistry; the levels of inflammatory factors i.e. interleukin-1β (1L-1β) and tumor necrosis factor-α (TNF-α) in the cartilage endplate were detected by ELISA; the protein expression of ADAMTS5, Aggrecan, matrix metalloproteinase-13 (MMP-13), IL-1β and TNF-α in the cartilage endplate was detected by Western blot.

RESULTS

Compared with the sham operation group, in the model group, the comprehensive response score was decreased (<0.01); L/L intervertebral disc and the cancellous bones of endplate vertebral body showed low signal and unclear boundary; the chondrocytes of the cartilage endplate increased significantly, the cells were enlarged and hypertrophic, and the nuclei were wrinkled and clustered; the positive expression of ADAMTS5 as well as the levels of IL-1β and TNF-α were increased (<0.01), while the positive expression of Aggrecan was decreased (<0.01) in the cartilage endplate; the protein expression of ADAMTS5, MMP-13, IL-1β and TNF-α was increased (<0.01), while that of Aggrecan was decreased (<0.01) in the cartilage endplate. Compared with the model group, in the EA group, the comprehensive response score was increased (<0.01); the signal of L/L intervertebral disc and the cancellous bones of endplate vertebral body was enhanced; the chondrocytes of the cartilage endplate were reduced, the nuclei were slightly crumpled and scattered; the positive expression of ADAMTS5 as well as the levels of IL-1β and TNF-α were decreased (<0.05, <0.01), while the positive expression of Aggrecan was increased (<0.01) in the cartilage endplate; the protein expression of ADAMTS5, MMP-13, IL-1β and TNF-α was decreased (<0.05, <0.01), while that of Aggrecan was increased (<0.05) in the cartilage endplate.

CONCLUSION

EA at "Jiaji" (EX-B 2) can delay the MC of cartilage endplate. The mechanism may be related to inhibiting the degradation of ECM of chondrocytes and the secretion of inflammatory factors, and repairing the degeneration of endplate cartilage.

摘要

目的

观察电针“夹脊”(EX - B 2)穴对软骨终板Modic改变(MC)家兔软骨细胞外基质(ECM)及炎症反应的影响,探讨电针治疗终板软骨MC的机制。

方法

将18只雄性新西兰白兔随机分为假手术组、模型组和电针组,每组6只。基于自身免疫理论,模型组和电针组家兔通过自体髓核植入建立MC模型。造模成功后,电针组于L₅、L₆双侧“夹脊”(EX - B 2)穴施针,采用疏密波,频率2 Hz/15 Hz,电流强度1 mA,每次20 min,每日1次,连续干预6 d后间隔1 d,共4周。观察造模前后及干预前后的综合反应评分。造模及干预后,采用磁共振成像(MRI)观察椎间盘及软骨终板信号强度。干预后,通过苏木精 - 伊红(HE)染色观察软骨终板软骨细胞形态;采用免疫组织化学法检测软骨终板中含血小板反应蛋白基序的解聚素和金属蛋白酶 - 5(ADAMTS5)及聚集蛋白聚糖的阳性表达;采用酶联免疫吸附测定(ELISA)法检测软骨终板中炎症因子白细胞介素 - 1β(IL - 1β)和肿瘤坏死因子 - α(TNF - α)水平;采用蛋白质印迹法检测软骨终板中ADAMTS5、聚集蛋白聚糖、基质金属蛋白酶 - 13(MMP - 13)、IL - 1β和TNF - α的蛋白表达。

结果

与假手术组比较,模型组综合反应评分降低(P < 0.01);L₅/L₆椎间盘及终板椎体松质骨呈低信号,边界不清;软骨终板软骨细胞显著增多,细胞增大、肥大,细胞核皱缩、聚集;软骨终板中ADAMTS5阳性表达及IL - 1β、TNF - α水平升高(P < 0.01),而聚集蛋白聚糖阳性表达降低(P < 0.01);软骨终板中ADAMTS5、MMP - 13、IL - 1β和TNF - α蛋白表达升高(P < 0.01),而聚集蛋白聚糖蛋白表达降低(P < 0.01)。与模型组比较,电针组综合反应评分升高(P < 0.01);L₅/L₆椎间盘及终板椎体松质骨信号增强;软骨终板软骨细胞减少,细胞核轻度皱缩、散在分布;软骨终板中ADAMTS5阳性表达及IL - 1β、TNF - α水平降低(P < 0.05,P < 0.01),而聚集蛋白聚糖阳性表达升高(P < 0.01);软骨终板中ADAMTS5、MMP - 13、IL - 1β和TNF - α蛋白表达降低(P < 0.05,P < 0.01),而聚集蛋白聚糖蛋白表达升高(P < 0.05)。

结论

电针“夹脊”(EX - B 2)穴可延缓软骨终板MC,其机制可能与抑制软骨细胞ECM降解及炎症因子分泌、修复终板软骨退变有关。

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