Netrin-1、Slit2在夹脊电针结合神经松动术调节兔坐骨神经损伤后Rho GTPases失衡的作用

doi:10.3969/j.issn.1006-9771.2022.08.007

Abstract

Abstract:

Objective To observe the effect of Jiaji electroacupuncture combined with neurodynamic mobilization on nerve conduction velocity and the expression of Netrin-1, Slit2 and Rho GTPases after sciatic nerve injury in rabbits.

Methods A total of 216 New Zealand rabbits were randomly divided into nomal control group (NC), vector virus group (VV), Jiaji electroacupuncture combined with neurodynamic mobilization group (EN), Netrin-1 group (N1), Slit2 group (S2) and N1+S2 group. Each group was divided into three subgroups according to the postoperative treatment time points (one, two and four weeks), with twelve rabbits in each subgroup. The rabbit model of Sunderland Ⅲ degree injury of the left sciatic nerve was established by clamping method. There was no intervention in NC, and virus was injected during the preparation of the model. Jiaji electroacupuncture and neurodynamic mobilization were administrated three days after operation in EN group. Nerve conduction velocity of sciatic nerve was measured with electromyography. The sciatic nerve and L_4-6 spinal cords were obtained, the expression of Netrin-1 and Slit2 mRNA were detected real-time quantitative polymerase chain reaction and immunofluorescence double staining, and the expression of Rac1, Cdc42 and RhoA protein were observed via Western blotting.

Results One, two and four weeks after modeling, the nerve conduction velocity was more in EN group and N1+S2 group than in N1 group and S2 group (P< 0.05); the Netrin-1 and Slit2 mRNA expression were higher in the EN group and N1+S2 group than in the VV group and NC group (P< 0.05); the protein expression of Rac1 and Cdc42 were higher, and the protein expression of RhoA was lower in EN group and N1+S2 group than in N1 group, S2 group and VV group (P< 0.05).

Conclusion Jiaji electroacupuncture combined with nerve mobilization may promote the axonal regeneration by regulating the expression of Netrin-1 and Slit2, adjusting the imbalance of Rho GTPases enzyme system, and then promoting cytoskeleton reorganization and peripheral nerve regeneration after injury.

Key words: sciatic nerve, Jiaji electroacupuncture combined with neurodymanic mobilization, Netrin-1, Slit2, Rho GTPase

CLC Number:

R745.4

ZHANG Li,WU Shanhong,ZHAO Lele,WANG Yan. Role of Netrin-1 and Slit2 in regulating the imbalance of Rho GTPases after Jiaji electroacupuncture combined with nerve mobilization for sciatic nerve injury in rabbits[J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2022, 28(8): 914-926.

Figures/Tables 17

References 30

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组别	n	术后1周	术后2周	术后4周
正常对照组	12	55.005±3.874	53.375±3.644	53.051±3.628
病毒空载组	12	40.105±3.145^a	29.865±3.956^a	27.465±3.953^a
夹脊电针结合神经松动术组	12	35.234±2.665^a,b	43.605±2.805^a,b	48.445±3.517^a,b
Netrin-1组	12	28.654±2.543^a,b,c	37.445±3.825^a,b,c	39.976±3.263^a,b,c
Slit2组	12	28.774±2.955^a,b,c	36.765±4.005^a,b,c	39.245±4.014^a,b,c
Netrin-1+Slit2腺病毒结合组	12	35.225±3.133^a,b,d,e	44.160±3.958^a,b,d,e	46.914±3.421^a,b,d,e
F值		120.507	56.166	73.892
P值		< 0.001	< 0.001	< 0.001

组别	n	术后1周	术后2周	术后4周
正常对照组	12	1.002±0.015	1.001±0.015	1.016±0.075
病毒空载组	12	1.265±0.111^a	1.415±0.206^a	1.525±0.284^a
夹脊电针结合神经松动术组	12	1.685±0.216^a,b	1.725±0.120^a,b	1.785±0.234^a,b
Netrin-1组	12	2.114±0.276^a,b,c	2.050±0.287^a,b,c	2.345±0.362^a,b,c
Slit2组	12	1.314±0.194^a,c,d	1.333±0.159^a,c,d	1.549±0.294^a,c,d
Netrin-1+Slit2腺病毒结合组	12	1.885±0.235^a,b,c,e	2.014±0.317^a,b,c,e	2.214±0.302^a,b,c,e
F值		55.633	46.449	37.734
P值		< 0.001	< 0.001	< 0.001

组别	n	术后1周	术后2周	术后4周
正常对照组	12	0.996±0.014	1.000±0.0124	1.004±0.085
病毒空载组	12	1.294±0.117^a	1.314±0.145^a	1.525±0.244
夹脊电针结合神经松动术组	12	1.665±0.234^a,b	1.814±0.254^a,b	1.915±0.243^a,b
Netrin-1组	12	2.284±0.267^a,b,c	2.455±0.383^a,b,c	2.585±0.364^a,b,c
Slit2组	12	1.245±0.184^a,c,d	1.264±0.163^a,c,d	1.479±0.226^a,c,d
Netrin-1+Slit2腺病毒结合组	12	2.125±0.294^a,b,c,e	2.265±0.274^a,b,c,e	2.454±0.313^a,b,c,e
F值		73.527	74.202	65.746
P值		< 0.001	< 0.001	< 0.001

组别	n	术后1周	术后2周	术后4周
正常对照组	12	0.971±0.088	0.977±0.089	1.009±0.113
病毒空载组	12	1.286±0.117^a	1.345±0.145^a	1.415±0.126^a
夹脊电针结合神经松动术组	12	1.585±0.165^a,b	1.614±0.165^a,b	1.654±0.147^a,b
Netrin-1组	12	1.325±0.150^a,c	1.337±0.135^a,c	1.465±0.135^a,c
Slit2组	12	1.877±0.185^a,b,c,d	1.885±0.183^a,b,c,d	1.945±0.225^a,b,c,d
Netrin-1+Slit2腺病毒结合组	12	1.914±0.206^a,b,c,d	1.935±0.188^a,b,c,d	1.954±0.250^a,b,c,d
F值		65.368	67.645	50.839
P值		< 0.001	< 0.001	< 0.001

组别	n	术后1周	术后2周	术后4周
正常对照组	12	0.984±0.075	0.996±0.083	1.012±0.055
病毒空载组	12	1.425±0.094^a	1.465±0.100^a	1.480±0.113^a
夹脊电针结合神经松动术组	12	1.685±0.132^a,b	1.745±0.192^a,b	1.765±0.249^a,b
Netrin-1组	12	1.364±0.084^a,c	1.394±0.105^a,c	1.406±0.116^a,c
Slit2组	12	2.374±0.273^a,b,c,d	2.464±0.315^a,b,c,d	2.475±0.327^a,b,c,d
Netrin-1+Slit2腺病毒结合组	12	2.354±0.235^a,b,c,d	2.415±0.342^a,b,c,d	2.425±0.315^a,b,c,d
F值		134.582	93.423	82.778
P值		< 0.001	< 0.001	< 0.001

Role of Netrin-1 and Slit2 in regulating the imbalance of Rho GTPases after Jiaji electroacupuncture combined with nerve mobilization for sciatic nerve injury in rabbits

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组别	n	术后1周	术后2周	术后4周
正常对照组	12	0.995±0.050	1.001±0.021	1.016±0.062
病毒空载组	12	0.425±0.033^a	0.354±0.043^a	0.296±0.044^a
夹脊电针结合神经松动术组	12	0.785±0.032^a,b	0.692±0.029^a,b	0.745±0.043^a,b
Netrin-1组	12	0.530±0.023^a,b,c	0.505±0.052^a,b,c	0.484±0.049^a,b,c
Slit2组	12	0.433±0.035^a,c,d	0.388±0.041^a,c,d	0.310±0.053^a,c,d
Netrin-1+Slit2腺病毒结合组	12	0.680±0.030^a,b,c,d,e	0.705±0.028^a,b,d,e	0.730±0.046^a,b,d,e
F值		490.986	530.711	352.238
P值		< 0.001	< 0.001	< 0.001

组别	n	术后1周	术后2周	术后4周
正常对照组	12	0.995±0.051	0.994±0.250	0.997±0.062
病毒空载组	12	0.322±0.072^a	0.240±0.010^a	0.225±0.035^a
夹脊电针结合神经松动术组	12	0.825±0.023^a,b	0.765±0.025^a,b	0.75±0.033^a,b
Netrin-1组	12	0.480±0.021^a,b,c	0.625±0.043^a,b,c	0.424±0.051^a,b,c
Slit2组	12	0.354±0.0832^a,c,d	0.215±0.013^a,c,d	0.201±0.021^a,c,d
Netrin-1+Slit2腺病毒结合组	12	0.717±0.052^a,b,c,d,e	0.794±0.038^a,b,d,e	0.615±0.042^a,b,c,d,e
F值		295.530	109.746	639.603
P值		< 0.001	< 0.001	< 0.001

组别	n	术后1周	术后2周	术后4周
正常对照组	12	0.995±0.060	0.995±0.040	1.002±0.035
病毒空载组	12	0.515±0.063^a	0.454±0.054^a	0.415±0.083^a
夹脊电针结合神经松动术组	12	0.874±0.100^a,b	0.754±0.113^a,b	0.760±0.153^a,b
Netrin-1组	12	0.677±0.052^a,b,c	0.620±0.065^a,b,c	0.585±0.073^a,b
Slit2组	12	0.530±0.110^a,c,d	0.406±0.083^a,c,d	0.375±0.076^a,c,d
Netrin-1+Slit2腺病毒结合组	12	0.745±0.120^a,b,c,d,e	0.908±0.140^a,b,c,d,e	0.877±0.092^a,b,c,d,e
F值		60.425	84.290	89.855
P值		< 0.001	< 0.001	< 0.001

组别	n	术后1周	术后2周	术后4周
正常对照组	12	1.031±0.092	1.027±0.110	0.985±0.120
病毒空载组	12	0.484±0.074^a	0.414±0.086^a	0.434±0.072^a
夹脊电针结合神经松动术组	12	0.848±0.050^a,b	0.734±0.023^a,b	0.834±0.150^a,b
Netrin-1组	12	0.631±0.012^a,b,c	0.620±0.011^a,b,c	0.607±0.013^a,b,c
Slit2组	12	0.525±0.070^a,c,d	0.374±0.065^a,c,d	0.394±0.120^a,c,d
Netrin-1+Slit2腺病毒结合组	12	0.727±0.073^a,b,c,d,e	0.837±0.010^a,b,c,d,e	0.725±0.085^a,b,c,d,e
F值		118.939	180.41	55.387
P值		< 0.001	< 0.001	< 0.001

组别	n	术后1周	术后2周	术后4周
正常对照组	12	1.011±0.302	1.000±0.287	0.995±0.153
病毒空载组	12	4.820±0.119^a	4.654±0.189^a	6.490±0.158^a
夹脊电针结合神经松动术组	12	1.837±0.129^a,b	1.452±0.0848^a,b	1.883±0.199^a,b
Netrin-1组	12	4.808±0.099^a,c	4.420±0.167^a,c	6.474±0.117^a,c
Slit2组	12	3.561±0.118^a,b,c,d	2.762±0.178^a,b,c,d	2.610±0.109^a,b,c,d
Netrin-1+Slit2腺病毒结合组	12	1.320±0.107^a,b,c,d,e	1.810±0.196^a,b,c,d,e	1.691±0.168^a,b,c,d,e
F值		1386.867	766.156	3130.581
P值		< 0.001	< 0.001	< 0.001

组别	n	术后1周	术后2周	术后4周
正常对照组	12	1.005±0.053	1.010±0.076	1.002±0.179
病毒空载组	12	2.421±0.214^a	2.714±0.254^a	3.142±0.318^a
夹脊电针结合神经松动术组	12	1.510±0.184^a,b	1.311±0.156^a,b	1.470±0.288^a,b
Netrin-1组	12	2.412±0.098^a,c	2.603±0.147^a,c	3.011±0.207^a,c
Slit2组	12	2.103±0.117^a,b,c,d	1.813±0.229^a,b,c,d	2.512±0.189^a,b,c,d
Netrin-1+Slit2腺病毒结合组	12	1.384±0.046^a,b,c,d,e	1.491±0.219^a,b,c,d,e	1.222±0.087^a,b,c,d,e
F值		232.092	162.279	236.881
P值		< 0.001	< 0.001	< 0.001

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