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福建中医药大学附属人民医院超声医学科,福州 350004
福建中医药大学附属人民医院康复医学科,福州 350004
福建中医药大学附属人民医院疼痛科,福州 350004
福建中医药大学附属人民医院病理科,福州 350004
✉陈斌,主治医师。E-mail:chenbin327425224@163.com
收稿:2024-06-01,
修回:2025-07-16,
网络首发:2025-07-16,
纸质出版:2025-10-12
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吴韦贤, 陈斌, 刘晶, 等. 基于Wnt/β-catenin信号通路探讨超声引导下椎间孔电针对脊髓损伤的影响[J]. 中国针灸, 2025,45(10):1442-1449.
WU Weixian, CHEN Bin, LIU Jing, et al. Effect of ultrasound-guided foraminal electroacupuncture on spinal cord injury based on the Wnt/β-catenin signaling pathway[J]. Chinese Acupuncture & Moxibustion, 2025, 45(10): 1442-1449.
吴韦贤, 陈斌, 刘晶, 等. 基于Wnt/β-catenin信号通路探讨超声引导下椎间孔电针对脊髓损伤的影响[J]. 中国针灸, 2025,45(10):1442-1449. DOI: 10.13703/j.0255-2930.20240601-k0004.
WU Weixian, CHEN Bin, LIU Jing, et al. Effect of ultrasound-guided foraminal electroacupuncture on spinal cord injury based on the Wnt/β-catenin signaling pathway[J]. Chinese Acupuncture & Moxibustion, 2025, 45(10): 1442-1449. DOI: 10.13703/j.0255-2930.20240601-k0004.
目的:
2
观察超声引导下椎间孔电针对脊髓损伤(SCI)模型大鼠神经细胞凋亡及运动功能的影响,探讨其可能作用机制。
方法:
2
将36只SPF级SD大鼠随机分为假手术组、模型组、超声引导下电针组,每组12只。假手术组暴露脊髓后即缝合伤口,不做打击;其余组均应用改良的Allen’s脊髓损伤打击法造模。造模成功后的第1、3、7、14天,超声引导下电针组在超声引导下于T
9
/T
10
、T
10
/T
11
椎间孔行电针干预,避免损伤脊髓,疏密波,频率2 Hz/100 Hz,电流1~2 mA。第1、3、7、14天干预结束后,评定大鼠Basso-Beattie-Bresnahan(BBB)评分,行斜板实验检测大鼠后肢抓力。干预结束后,采用HE染色法观察大鼠脊髓形态,TUNEL染色法检测大鼠脊髓神经细胞凋亡情况,ELISA法检测大鼠血清白细胞介素(IL)-6、IL-1β、肿瘤坏死因子-α(TNF-α)含量,Western blot法检测大鼠脊髓无翼型相关整合位点-4(Wnt-4)、β-连环蛋白(β-catenin)、细胞髓细胞瘤病毒原癌基因同源物(c-Myc)、B细胞淋巴瘤-2(Bcl-2)相关X蛋白(Bax)、Bcl-2、神经元核(NeuN)蛋白表达,实时荧光定量PCR法检测大鼠脊髓Wnt-4、β-catenin、c-Myc、Bax、Bcl-2、NeuN mRNA表达。
结果:
2
与假手术组比较,模型组各时点BBB评分降低(
P
<
0.05),斜板角度减小(
P
<
0.05);与模型组比较,超声引导下电针组第3、7、14天干预结束后BBB评分升高(
P
<
0.05),第1、3、7、14天干预结束后斜板角度增加(
P
<
0.05)。与假手术组比较,模型组脊髓结构紊乱,炎性细胞、坏死神经元增多;脊髓凋亡神经细胞数量增多(
P
<
0.05);血清IL-6、IL-1β、TNF-α含量升高(
P
<
0.05);脊髓Wnt-4、β-catenin、c-Myc、Bax蛋白及mRNA表达升高(
P
<
0.05),Bcl-2、NeuN蛋白及mRNA表达降低(
P
<
0.05)。与模型组比较,超声引导下电针组脊髓炎性细胞减少;凋亡神经细胞数量减少(
P
<
0.05);血清IL-6、IL-1β、TNF-α含量降低(
P
<
0.05);脊髓Wnt-4、β-catenin、Bcl-2、NeuN蛋白及mRNA表达升高(
P
<
0.05),c-Myc、Bax蛋白及mRNA表达降低(
P
<
0.05)。
结论:
2
超声引导下椎间孔电针可改善SCI模型大鼠运动功能,可能通过调控Wnt-4/β-catenin信号通路相关分子表达,抑制神经细胞凋亡和炎症反应实现。
Objective
2
To observe the effects of ultrasound-guided foraminal electroacupuncture on neuronal apoptosis and motor function in rats with spinal cord injury (SCI)
and to explore the potential underlying mechanisms.
Methods
2
Thirty-six SPF-grade Sprague-Dawley rats were randomly assigned to a sham operation group
a model group
and an ultrasound-guilded electroacupuncture group (electroacupuncture group)
with 12 rats in each group. In the sham operation group
the spinal cord was exposed and then t
he incision was sutured without contusion. In the other two groups
SCI models were established using a modified Allen’s impact method. On days 1
3
7
and 14 after modeling
the electroacupuncture group received electroacupuncture intervention at the T
9
/T
10
and T
10
/T
11
intervertebral foramen under ultrasound guidance
avoiding spinal cord injury. Stimulation parameters were dense-disperse wave at 2 Hz/100 Hz and 1-2 mA for each session. Following interventions on days 1
3
7
and 14
the Basso-Beattie-Bresnahan (BBB) score was assessed; the inclined plane test was used to assess hindlimb grip strength in rats. After the intervention
HE staining was used to observe spinal cord morphology; TUNEL staining was used to detect neuronal apoptosis; ELISA was used to measure the serum levels of interleukin (IL)-6
IL-1β
and tumor necrosis factor-alpha (TNF-α); Western blot was used to analyze the protein expression of Wnt-4
β-catenin
c-Myc
Bax
Bcl-2
and NeuN in spinal tissue; quantitative real-time PCR was used to detect the mRNA expression of Wnt-4
β-catenin
c-Myc
Bax
Bcl-2
and NeuN.
Results
2
Compared with the sham operation group
the model group showed significantly reduced BBB scores (
P
<
0.05)
and reduced inclined plane angles (
P
<
0.05) at all time points. Compared with the model group
the electroacupuncture group exhibited increased BBB scores on days 3
7
and 14 (
P
<
0.05)
and higher inclined plane angles on days 1
3
7
and 14 (
P
<
0.05). Compared with the sham operation group
the model group showed disorganized spinal cord structure with increased inflammatory cells and necrotic neurons
higher number of apoptotic neurons in spinal tissue (
P
<
0.05)
elevated serum IL-6
IL-1β
and TNF-α levels (
P
<
0.05)
increased protein and mRNA expression of Wnt-4
β-catenin
c-Myc
and Bax (
P
<
0.05)
and
decreased protein and mRNA expression of Bcl-2 and NeuN in spinal tissue (
P
<
0.05). Compared with the model group
the electroacupuncture group had fewer inflammatory cells and apoptotic neurons in spinal tissue (
P
<
0.05)
reduced serum IL-6
IL-1β
and TNF-α levels (
P
<
0.05)
increased protein and mRNA expression of Wnt-4
β-catenin
Bcl-2
and NeuN (
P
<
0.05)
and decreased protein and mRNA expression of c-Myc and Bax in spinal tissue (
P
<
0.05).
Conclusion
2
Ultrasound-guided foraminal electroacupuncture could improve motor function in rats with SCI
potentially by regulating the expression of molecules related to the Wnt-4/β-catenin signaling pathway to inhibit neuronal apoptosis and inflammatory responses.
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