Sun Jiawei, Zhang Yingqi, Liu Zhifeng, Zhang Hanyu, Liu Jiayue, Xu Yue, Na Rentuya, Zhang Hongzheng, Yan Jiawang, Yu Tianyuan
School of Acupuncture-Moxibustion and Tuina, Beijing University of Chinese Medicine, Beijing, China.
Department of Tuina and Pain Management, Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing, China.
Front Neurol. 2025 Jul 30;16:1622602. doi: 10.3389/fneur.2025.1622602. eCollection 2025.
To investigate whether Tuina therapy alleviated inflammation and motor neuron apoptosis in sciatic nerve injury (SNI) rats by regulating cytosolic phospholipase A2 (cPLA2) and Ras homolog family member A/Rho-associated coiled-coil comprising protein kinase 2 (RhoA/ROCK2) signaling cascades.
Four experimental cohorts were established utilizing 36 male Sprague-Dawley rats: control, sham, SNI, and TUI. We implemented a sciatic nerve injury (SNI) model. At dthe mid-thigh level, sciatic nerves were exposed and crushed for 5 s using non-serrated forceps at points spaced approximately 2 mm apart. Postoperatively, Tuina therapy (Chinese therapeutic massage, Tuina) was administered to evaluate its neuromodulatory effects. SNI models were established in the SNI and TUI cohorts. TUI cohorts applied with "Three-Manipulation and Three-Acupoint" technique, which included pressing, plucking, and kneading on the acupoints Yinmen (BL37), Chengshan (BL57), and Yanglingquan (GB34). The control cohort underwent no intervention. The sham surgery and model cohorts underwent restraining interventions. Motor function was assessed using Basso, Beattie, and Bresnahan (BBB) scores and CatWalk gait analysis. Spinal cord (SC) histology was evaluated using hematoxylin and eosin and Nissl staining. NeuN-positive cells were quantified via immunofluorescence. Tumor necrosis factor- (TNF-α), interleukin-6 (IL-6), and aquaporin-4 levels were determined through enzyme-linked immunosorbent assay. RhoA, ROCK2, Bax, Bcl-2, and cPLA2 mRNA levels were analyzed using real-time quantitative polymerase chain reaction. RhoA, ROCK2, Bax, Bcl-2, cPLA2, and p-cPLA2 protein expressions were analyzed using western blotting to investigate the impact of Tuina therapy on nerve regeneration and apoptosis regulation.
The TUI cohort showed better BBB scores and CatWalk results than the SNI cohort (all < 0.001). Histological analysis revealed diminished inflammatory cell infiltration and increased neuronal survival. NeuN immunofluorescence indicated decreased motor neuron apoptosis in the anterior horn of the SC. Tuina therapy reversed TNF-, IL-6, and aquaporin-4 levels ( < 0.01). The TUI cohort had lower mRNA expression of Bax, cPLA2, and ROCK2 (all 0.001), mRNA expression of RhoA ( 0.01), and Bax, cPLA2, p-cPLA2, and RhoA/ROCK2 levels (all 0.001) than the SNI cohort. Conversely, mRNA and protein expression levels of Bcl2 were higher in the TUI cohort than in the SNI cohort (all 0.001).
Tuina therapy improved motor function in SNI rats by inhibiting motor neuron apoptosis via cPLA2 regulation, potentially via the RhoA/ROCK2 signaling pathway.
探讨推拿疗法是否通过调节胞质型磷脂酶A2（cPLA2）和Ras同源家族成员A/ Rho相关卷曲螺旋形成蛋白激酶2（RhoA/ROCK2）信号级联反应，减轻坐骨神经损伤（SNI）大鼠的炎症反应和运动神经元凋亡。
利用36只雄性Sprague-Dawley大鼠建立四个实验队列：对照组、假手术组、SNI组和推拿组。我们建立了坐骨神经损伤（SNI）模型。在大腿中部水平，暴露坐骨神经，并用无齿镊在相距约2 mm的点处夹捏5秒。术后，给予推拿疗法（中医推拿）以评估其神经调节作用。在SNI组和推拿组中建立SNI模型。推拿组采用“三法三穴”技术，包括按压、弹拨和揉按穴位殷门（BL37）、承山（BL57）和阳陵泉（GB34）。对照组不进行干预。假手术组和模型组进行约束干预。使用Basso、Beattie和Bresnahan（BBB）评分和CatWalk步态分析评估运动功能。使用苏木精-伊红染色和尼氏染色评估脊髓（SC）组织学。通过免疫荧光对NeuN阳性细胞进行定量。通过酶联免疫吸附测定法测定肿瘤坏死因子-（TNF-α）、白细胞介素-6（IL-6）和水通道蛋白-4水平。使用实时定量聚合酶链反应分析RhoA、ROCK2、Bax、Bcl-2和cPLA2 mRNA水平。使用蛋白质印迹法分析RhoA、ROCK2、Bax、Bcl-2、cPLA2和p-cPLA2蛋白表达，以研究推拿疗法对神经再生和凋亡调节的影响。
推拿组的BBB评分和CatWalk结果优于SNI组（均P<0.001）。组织学分析显示炎症细胞浸润减少，神经元存活增加。NeuN免疫荧光表明脊髓前角运动神经元凋亡减少。推拿疗法使TNF-、IL-6和水通道蛋白-4水平逆转（P<0.01）。推拿组的Bax、cPLA2和ROCK2 mRNA表达较低（均P<0.001），RhoA mRNA表达较低（P<0.01），Bax、cPLA2、p-cPLA2和RhoA/ROCK2水平较低（均P<0.001），均低于SNI组。相反，推拿组的Bcl2 mRNA和蛋白表达水平高于SNI组（均P<0.001）。
推拿疗法通过调节cPLA2抑制运动神经元凋亡，可能通过RhoA/ROCK2信号通路，改善SNI大鼠的运动功能。
