“三法三穴”推拿手法对坐骨神经损伤大鼠运动功能和转化生长因子β1/Smad2通路蛋白表达的影响

doi:10.3969/j.issn.1006-9771.2020.04.009

摘要/Abstract

摘要：

目的探讨“三法三穴”推拿手法对坐骨神经损伤大鼠后肢运动功能,施万细胞增殖、髓鞘修复以及坐骨神经中转化生长因子β1 (TGF-β1)/Smad2通路蛋白表达的影响。方法 66只雄性Sprague-Dawley大鼠随机分为假手术组(n = 22)、模型组(n = 22)和观察组(n = 22)。模型组和观察组采用夹持法制备坐骨神经损伤模型。造模后第8天,观察组每天以点法、拨法、揉法定性定量刺激大鼠术侧殷门、承山、阳陵泉。分别于干预前和干预21 d测量坐骨神经功能指数(SFI);干预前,干预7 d、14 d、21 d行斜板测试;干预21 d,免疫荧光染色观察S100、TGF-β1、Smad2的表达;干预前,干预7 d、21 d,Western blotting检测TGF-β1、Smad2和p-Smad2蛋白表达。结果干预前,模型组和观察组SFI和斜板测试角度低于假手术组(P < 0.05);干预21 d,观察组SFI和斜板测试角度均高于模型组( P< 0.05)。免疫荧光染色显示,干预21 d,模型组坐骨神经S100表达明显低于假手术组(P < 0.01),观察组高于模型组( P < 0.05),且与假手术组比较无显著性差异( P > 0.05)。Western blotting显示,干预前和干预7 d,模型组TGF-β1、Smad2和p-Smad2表达较假手术组升高( P < 0.05);干预21 d,三组间各蛋白表达均无显著性差异( P > 0.05)。结论 “三法三穴”推拿手法能促进施万细胞增殖,促进髓鞘恢复,改善坐骨神经损伤大鼠后肢运动功能,但该作用可能与TGF-β1/Smad2通路无关。

关键词: 周围神经损伤, 推拿, 髓鞘, 大鼠

Abstract:

Objective To investigate the effect of Tuina of Three Handing-Three Points on the motor function of hind limbs, the proliferation of Schwann cell, recovery of myelin sheath and the expression of transforming growth factor (TGF)-β1/Smad2 pathway protein in injured sciatic nerve of rats.Methods A total of 66 male Sprague-Dawley rats were randomly divided into sham operation group (n = 22), model group (n= 22) and observation group (n= 22). The sciatic nerve injury model was made by clamping method. On the eighth day after modeling, the observation group received stimulation on Yinmen (BL37), Chengshan (BL57) and Yanglingquan (GB34). The sciatic functional index (SFI) was measured before intervention and 21 days after intervention. The Oblique Plate Test was performed before intervention, and seven days, 14 days and 21 days after intervention. The expression of S100, TGF-β1 and Smad2 were observed by immunofluorescence. The expression of TGF-β1, Smad2 and p-Smad2 was detected by Western blotting.Results Before intervention, SFI was lower in the model group and observation group than in the sham operation group (P < 0.05); 21 days after intervention, SFI and the angle of Oblique Plate Test were higher in the observation group than in the model group ( P < 0.05). Immunofluorescence showed that, 21 days after intervention, the expression of S100 was lower in the model group than in the sham operation group ( P < 0.01), and was higher in the observation group than in the model group ( P < 0.05), no difference was found between the observation group and the sham operation group ( P > 0.05). Western blotting showed that, before intervention and seven days after intervention, the expression of TGF-β1, Smad2 and p-Smad2 were higher in the model group than in the sham operation group; 21 days after intervention, no difference was found in the expression among groups ( P > 0.05) Conclusion Three Handing-Three Points could promote the proliferation of Schwann cell and the recovery of myelin sheath, to improve the motor function of hind limbs in rats with sciatic nerve injury, which may not be related to TGF-β1/Smad2 pathway.

Key words: peripheral nerve injury, Tuina, myelin sheath, rats

中图分类号:

R745.4

邵帅,莫岩君,于天源,沈熠,罗宇婷,张羽墨,吕桃桃. “三法三穴”推拿手法对坐骨神经损伤大鼠运动功能和转化生长因子β1/Smad2通路蛋白表达的影响[J]. 《中国康复理论与实践》, 2020, 26(4): 432-439.

SHAO Shuai,MO Yan-jun,YU Tian-yuan,SHEN Yi,LUO Yu-ting,ZHANG Yu-mo,LÜ Tao-tao. Effects of Three Handing-Three Points on Motor Function and Expression of Transforming Growth Factor-β1/Smad2 Pathway in Injured Sciatic Nerve of Rats[J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2020, 26(4): 432-439.

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组别	n	干预前	干预21 d
假手术组	6	-6.18±8.26	-4.78±4.82
模型组	6	-100±0^a	-26.67±9.51^a
观察组	6	-100±0^a	-6.21±7.41^b
F值		31.320	16.000
P值		< 0.001	< 0.001

组别	n	干预前	干预7 d	干预14 d	干预21 d
假手术组	6	42.08±2.64	42.83±0.69	43.07±2.12	41.69±1.29
模型组	6	36.64±1.39^a	36.00±1.00^a	37.23±2.84^a	37.17±1.87^a
观察组	6	37.03±2.80^a	36.04±1.72^a	38.02±2.73^a	39.84±1.45^a,b
F值		9.210	81.700	9.040	12.790
P值		0.002	< 0.001	0.003	< 0.001

组别	n	TGF-β1	Smad2	S100
假手术组	4	223.00±10.90	205.63±19.13	204.37±9.10
模型组	4	225.46±15.72	192.71±20.73	188.36±7.13^a
观察组	4	219.83±15.43	209.36±15.40	209.87±2.65^b
F值		0.160	0.880	10.640
P值		0.866	0.445	0.004

组别	n	干预前	干预7 d	干预21 d
假手术组	6	0.90±0.30	1.11±0.24	0.86±0.20
模型组	6	2.09±0.61^a	1.96±0.56^a	0.79±0.16
观察组	6		1.55±0.74	0.84±0.22
F值		18.650	3.520	3.220
P值		0.002	0.056	0.069

组别	n	干预前	干预7 d	干预21 d
假手术组	6	1.33±0.63	1.01±0.51	0.94±0.20
模型组	6	4.78±0.82^a	2.45±0.68^a	1.40±0.81
观察组	6		1.60±0.54	1.35±0.31
F值		66.970	9.450	1.420
P值		< 0.001	0.002	0.273