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1.湖北中医药大学针灸骨伤学院,武汉 430065
2.武汉市中西医结合医院针灸科
3.深圳市龙岗区妇幼保健院/汕头大学医学院龙岗妇幼临床学院,广东 深圳 518172
汪敏,湖北中医药大学博士研究生。E-mail:1198692128@qq.com
☒黄国付,主任医师。E-mail: 531529128@qq.com
收稿:2023-09-28,
网络首发:2024-07-22,
纸质出版:2024-09-12
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汪敏, 邹璟, 黄国付. 电针“夹脊”穴对软骨终板Modic改变模型兔软骨细胞外基质和炎性反应的影响[J]. 中国针灸, 2024,44(9):1046-1053.
WANG Min, ZOU Jing, HUANG Guofu. Effects of electroacupuncture at "Jiaji" (EX-B 2) on extracellular matrix of chondrocytes and inflammatory reaction in rabbits with Modic changes of cartilage endplate[J]. Chinese Acupuncture & Moxibustion, 2024, 44(9): 1046-1053.
汪敏, 邹璟, 黄国付. 电针“夹脊”穴对软骨终板Modic改变模型兔软骨细胞外基质和炎性反应的影响[J]. 中国针灸, 2024,44(9):1046-1053. DOI: 10.13703/j.0255-2930.20230928-k0002.
WANG Min, ZOU Jing, HUANG Guofu. Effects of electroacupuncture at "Jiaji" (EX-B 2) on extracellular matrix of chondrocytes and inflammatory reaction in rabbits with Modic changes of cartilage endplate[J]. Chinese Acupuncture & Moxibustion, 2024, 44(9): 1046-1053. DOI: 10.13703/j.0255-2930.20230928-k0002.
目的:
2
观察电针“夹脊”穴对软骨终板Modic改变(MC)模型兔软骨细胞外基质(ECM)和炎性反应的影响,探讨电针治疗软骨终板MC的作用机制。
方法:
2
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。模型组和电针组实验兔基于自身免疫学说采用自体髓核填充法制备MC模型。造模成功后电针组实验兔予电针双侧L
5
、L
6
“夹脊”穴干预,疏密波,频率2 Hz/15 Hz,电流强度1 mA,每次20 min,每天1次,连续干预6 d后休息1 d,共干预4周。于造模前后及干预前后观察各组实验兔综合反应评分;于造模后和干预后采用磁共振成像(MRI)观察实验兔椎间盘及软骨终板信号强度;干预后,采用HE染色观察实验兔软骨终板的软骨细胞形态,免疫组化法检测实验兔软骨终板血小板反应蛋白解整合素金属肽酶5(ADAMTS5)和聚集蛋白聚糖(Aggrecan)阳性表达,ELISA法检测实验兔软骨终板炎性因子[白细胞介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)
]
含量,Western blot法检测实验兔软骨终板ADAMTS5、Aggrecan、基质金属蛋白酶-13(MMP-13)、IL-1β和TNF-α蛋白表达。
结果:
2
与假手术组比较,模型组实验兔综合反应评分降低(
P
<
0.01);L
5
/L
6
椎间盘及软骨终板下椎体松质骨出现低信号,分界不清;软骨终板的软骨细胞明显增多,细胞肿大而肥厚,细胞核皱缩、坏死,聚集排列;软骨终板ADAMTS5阳性表达及IL-1β、TNF-α含量升高(
P
<
0.01),Aggrecan阳性表达降低(
P
<
0.01);软骨终板ADAMTS5、MMP-13、IL-1β和TNF-α蛋白表达升高(
P
<
0.01),Aggrecan蛋白表达降低(
P
<
0.01)。与模型组比较,电针组实验兔综合反应评分升高(
P
<
0.01);L
5
/L
6
椎间盘及软骨终板下椎体松质骨信号增强;软骨终板的软骨细胞减少,细胞核轻度皱缩,散在分布;软骨终板ADAMTS5阳性表达及IL-1β和TNF-α含量降低(
P
<
0.05,
P
<
0.01),Aggrecan阳性表达升高(
P
<
0.01);软骨终板ADAMTS5、MMP-13、IL-1β和TNF-α蛋白表达降低(
P
<
0.05,
P
<
0.01),Aggrecan蛋白表达升高(
P
<
0.05)。
结论:
2
电针“夹脊”穴能延缓软骨终板MC,其机制可能与抑制软骨细胞ECM降解和炎性因子分泌、修复变性的软骨终板有关。
Objective
2
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
2
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
5
and L
6
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
2
Compared with the sham operation group
in the model group
the comprehensive response score was decreased (
P
<
0.01); L
5
/L
6
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 (
P
<
0.01)
while the positive expression of Aggrecan was decreased (
P
<
0.01) in the cartilage endplate; the protein expression of ADAMTS5
MMP-13
IL-1β and TNF-α was increased (
P
<
0.01)
while that of Aggrecan was decreased (
P
<
0.01) in the cartilage endplate. Compared with the model group
in the EA group
the comprehensive response score was increased (
P
<
0.01); the signal of L
5
/L
6
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 (
P
<
0.05
P
<
0.01)
while the positive expression of Aggrecan was increased (
P
<
0.01) in the cartilage endplate; the protein expression of ADAMTS5
MMP-13
IL-1β and TNF-α was decreased (
P
<
0.05
P
<
0.01)
while that of Aggrecan was increased (
P
<
0.05) in the cartilage endplate.
Conclusion
2
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.
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