The Role and Mechanism of Aspirin Combined with Rehabilitation Training in the Repair of Sciatic Nerve Injury and the Changes in the Schwannocytus (Schwann Cells) in Rats / Rol y Mecanismo de la Aspirina Combinados con Entrenamiento de Rehabilitación en la Reparación de la Lesión del Nervio Ciático y los Cambios en los Schwannocitos (Células de Schwann) en Ratas

SUMMARY: This study investigated the role and mechanism of aspirin combined with rehabilitation training in the nerve injury repair and Schwann cell changes in rats with sciatic nerve injury. Totally, 120 male healthy SD rats were randomly divided into sham, model, aspirin, and aspirin + rehabilitation groups, with 30 rats in each group. The sciatic nerve function index (SFI), photothermal pain tolerance threshold and inclined plane test results at 4, 6, and 8 weeks after operation were compared. The distance of sensory nerve regeneration and the expression of S100B protein in Schwann cells were analyzed. Compared with the sham group, the SFI of the model, aspirin, and aspirin+rehabilitation groups were significantly lower at 4, 6, and 8 weeks after operation. However, the aspirin and aspirin+rehabilitation groups had significantly higher SFI than the model group. The SFI at 6 and 8 weeks after operation was higher in the aspirin+rehabilitation group than that in the aspirin group (P<0.05). The photothermal pain tolerance threshold of the sham, aspirin, and aspirin+rehabilitation groups were significantly higher than those of the model group at 4, 6, and 8 weeks after operation (P<0.05). The inclination angles of the model, aspirin, and aspirin+rehabilitation groups were significantly lower than those of the sham group at 4, 6, and 8 weeks after operation, and the inclination angle of the aspirin+rehabilitation group was significantly higher than that of the model and aspirin groups (P<0.05). The sensory nerve regeneration distance in aspirin and aspirin+rehabilitation groups was higher than that in the sham and model groups (P<0.05). The expression of S100B protein in the aspirin and aspirin+rehabilitation groups was higher than that in the model group (P<0.05). Aspirin combined with rehabilitation training can promote the functional recovery of sciatic nerve injury, and the mechanism may be related to the increase of the expression of S100B protein in Schwann cells.

En este estudio se investigó el papel y el mecanismo que desempeña la aspirina combinada, con el entrenamiento de rehabilitación en la reparación de lesiones nerviosas y los cambios en los schwannocitos en ratas con lesiones en el nervio ciático. En total, 120 ratas SD macho sanas se dividieron aleatoriamente en cuatro grupos de 30 ratas en cada uno: simulación, modelo, aspirina y aspirina + rehabilitación. Se compararon el índice de función del nervio ciático (SFI), el umbral de tolerancia al dolor fototérmico y los resultados de la prueba del plano inclinado a las 4, 6 y 8 semanas después de la operación. Se analizó la distancia de regeneración del nervio sensorial y la expresión de la proteína S100B en los schwannocitos. En comparación con el grupo simulado, el SFI de los grupos modelo, aspirina y aspirina+rehabilitación fue significativamente menor a las 4, 6 y 8 semanas después de la operación. Sin embargo, los grupos de aspirina y aspirina + rehabilitación tuvieron un SFI significativamente más alto que el grupo modelo. El SFI a las 6 y 8 semanas después de la operación fue mayor en el grupo de aspirina + rehabilitación que en el grupo de aspirina (P<0,05). El umbral de tolerancia al dolor fototérmico de los grupos simulado, aspirina y aspirina+rehabilitación fue significativamente mayor que el del grupo modelo a las 4, 6 y 8 semanas después de la operación (P<0,05). Los ángulos de inclinación de los grupos modelo, aspirina y aspirina+rehabilitación fueron significativamente menores que los del grupo simulado a las 4, 6 y 8 semanas después de la operación, y el ángulo de inclinación del grupo aspirina+rehabilitación fue significativamente mayor que el de los grupos modelo y aspirina (P<0.05). La distancia de regeneración del nervio sensorial en los grupos de aspirina y aspirina+rehabilitación fue mayor que en los grupos simulado y modelo (P<0,05). La expresión de la proteína S100B en los grupos de aspirina y aspirina+rehabilitación fue mayor que en el grupo modelo (P<0,05). La aspirina combinada con el entrenamiento de rehabilitación puede promover la recuperación funcional de la lesión del nervio ciático, y el mecanismo puede estar relacionado con el aumento de la expresión de la proteína S100B en los schwannocitos.

Comparison of the Nerve Regeneration Capacity and Characteristics between Sciatic Nerve Crush and Transection Injury Models in Rats / 生物医学与环境科学（英文）

OBJECTIVE@#To provide useful information for selecting the most appropriate peripheral nerve injury model for different research purposes in nerve injury and repair studies, and to compare nerve regeneration capacity and characteristics between them.@*METHODS@#Sixty adult SD rats were randomly divided into two groups and underwent crush injury alone (group A, n = 30) or transection injury followed by surgical repair (group B, n = 30) of the right hind paw. Each group was subjected to the CatWalk test, gastrocnemius muscle evaluation, pain threshold measurement, electrophysiological examination, retrograde neuronal labeling, and quantification of nerve regeneration before and 7, 14, 21, and 28 days after injury.@*RESULTS@#Gait analysis showed that the recovery speed in group A was significantly faster than that in group B at 14 days. At 21 days, the compound muscle action potential of the gastrocnemius muscle in group A was significantly higher than that in group B, and the number of labeled motor neurons in group B was lower than that in group A. The number of new myelin sheaths and the g-ratio were higher in group A than in group B. There was a 7-day time difference in the regeneration rate between the two injury groups.@*CONCLUSION@#The regeneration of nerve fibers was rapid after crush nerve injury, whereas the transection injury was relatively slow, which provides some ideas for the selection of clinical research models.

Local vibration therapy promotes the recovery of nerve function in rats with sciatic nerve injury / 中西医结合学报

OBJECTIVE@#It has been reported that local vibration therapy can benefit recovery after peripheral nerve injury, but the optimized parameters and effective mechanism were unclear. In the present study, we investigated the effect of local vibration therapy of different amplitudes on the recovery of nerve function in rats with sciatic nerve injury (SNI).@*METHODS@#Adult male Sprague-Dawley rats were subjected to SNI and then randomly divided into 5 groups: sham group, SNI group, SNI + A-1 mm group, SNI + A-2 mm group, and SNI + A-4 mm group (A refers to the amplitude; n = 10 per group). Starting on the 7th day after model initiation, local vibration therapy was given for 21 consecutive days with a frequency of 10 Hz and an amplitude of 1, 2 or 4 mm for 5 min. The sciatic function index (SFI) was assessed before surgery and on the 7th, 14th, 21st and 28th days after surgery. Tissues were harvested on the 28th day after surgery for morphological, immunofluorescence and Western blot analysis.@*RESULTS@#Compared with the SNI group, on the 28th day after surgery, the SFIs of the treatment groups were increased; the difference in the SNI + A-2 mm group was the most obvious (95% confidence interval [CI]: [5.86, 27.09], P < 0.001), and the cross-sectional areas of myocytes in all of the treatment groups were improved. The G-ratios in the SNI + A-1 mm group and SNI + A-2 mm group were reduced significantly (95% CI: [-0.12, -0.02], P = 0.007; 95% CI: [-0.15, -0.06], P < 0.001). In addition, the expressions of S100 and nerve growth factor proteins in the treatment groups were increased; the phosphorylation expressions of ERK1/2 protein in the SNI + A-2 mm group and SNI + A-4 mm group were upregulated (95% CI: [0.03, 0.96], P = 0.038; 95% CI: [0.01, 0.94], P = 0.047, respectively), and the phosphorylation expression of Akt in the SNI + A-1 mm group was upregulated (95% CI: [0.11, 2.07], P = 0.031).@*CONCLUSION@#Local vibration therapy, especially with medium amplitude, was able to promote the recovery of nerve function in rats with SNI; this result was linked to the proliferation of Schwann cells and the activation of the ERK1/2 and Akt signaling pathways.

The Effect of Anakinra on Acrylamide-induced Peripheral Neuropathy and Neuropathic Pain in Rats

Abstract Acrylamide is a neurotoxic compound. Moreover, anakinra is an interleukin-1 (IL-1) receptor antagonist used in rheumatoid arthritis treatment. This study investigated the effect of anakinra on acrylamide-related neuropathy and neuropathic pain. Acrylamide exposure caused a significant decrease in the pain threshold; an increase in malondialdehyde (MDA), tumor necrosis factor-alpha (TNF-α), and interleukin-1 beta (IL-1ß) levels; and a decrease in total glutathione (tGSH) values in the sciatic nerve. This indicates hyperalgesia presence, oxidative stress, and peripheral nerve tissue inflammation. Anakinra treatment significantly reduced the MDA, IL-1ß, and TNF-α levels, and increased the pain threshold and mean tGSH values. The analgesic effect of anakinra was 67.9% at the first hour, increasing to 74.9% and 76.7% at the second and third hours, respectively. The group receiving acrylamide exhibited histopathological changes (e.g., swollen and degenerated axons, hypertrophic and hyperplasic Schwann cells, and congested vessels). The use of anakinra significantly improved these morphological changes. Anakinra is concluded to reduce neuropathic pain and prevent neurotoxic effect of acrylamide on peripheral nerves due to its analgesic, antioxidant, and anti-inflammatory properties

A morphological study of denervation and reinnervation of neuromuscular junction and muscle spindle after sciatic nerve crush in mice / 中华显微外科杂志

Objective:To observe the denervation and reinnervation of the neuromuscular junction (NMJ) and muscle spindle after sciatic nerve crush in mice.Methods:From January, 2019 to October, 2019, 18 C57BL/6 mice were randomly and evenly divided into injured groups (sciatic nerve crush group, 12 mice) and control groups (sham group, 6 mice). Mice in the injured group were sacrificed at day-1, day-2, day-3 and 4 weeks after the sciatic nerve injury. Mice in the control group were sacrificed at day-3 and 4 weeks from the day of experiment. The tibialis anterior muscles with crushed nerve were collected for immunohistochemistry staining by neurofilament (NF), synaptophysin (Syn) and α-bungarotoxin (α-BTX). Data were analysed by unpaired t-test. P<0.05 was considered significant difference. Results:The completely denervation of NMJ reached (92.4±8.85)% at day-3 after the nerve crush, compared with (5.19±1.32)% in the control group ( P<0.05). However, the muscle spindle lost γ-motor neuron innervation at day-2 after the nerve crush and completely denervated at day-3 after the surgery. For the reinnervation, no difference of no reinnervation of NMJ [(3.02±0.78)% vs. (4.22±2.08)%], partial reinnervation of NMJ [(6.44±1.91)% vs. (7.94±2.12)%] and completely reinnervation of NMJ [(90.54±10.44)% vs. (87.84±13.89)%] were observed between the control group and the injured group 4 weeks after the injury. While, for the muscle spindle, only the acetylcholine receptors at each end were partly reinnervated. The innervation of the middle part of the muscle spindle was thin and discontinued without typical spiral winding structure. Conclusion:The denervation speed of NMJ and muscle spindle was comparable after the peripheral nerve injury. The γ-motor neuron lost innervation ahead of the sensory neuron in the muscle spindle. The reinnervation of NMJ is earlier than that of the muscle spindle, and the re-innervation of the γ-motor neuron is prior to the sensory neuron in the muscle spindle.

Effect of Three Handing-Three Points on Pain Function and Expression of CX3CL1/CX3CR1 in Spinal Cord Dorsal Horn of Rats with Sciatic Nerve Injury / 中国康复理论与实践

Objective:To investigate the mechanism of Three Handing-Three Points on pain function in sciatic nerve injury rats by observing the changes of chemokine (C-X3-C motif) ligand 1, CX3CL1)/chemokine (C-X3-C motif) receptor 1 (CX3CR1) protein and mRNA expression in spinal dorsal horn. Methods:A total of 74 male Sprague-Dawley rats were randomly divided into normal group (n= 12), sham group (n = 24), model group (n = 25), and Three Handing-Three Points group (Tuina group,n = 13). The model group and Tuina group prepared the sciatic nerve injury model. The sham group exposed sciatic nerve only. Tuina group received Tuina on Yinmen (BL37), Chengshan (BL57) and Yanglingquan (GB34) with Tuina manipulation emulator. The photothermal pain threshold was measured seven days after modeling and after 20 days of intervention; cumulative pain score was measured seven days after modeling, and after ten days and 20 days of intervention. The spinal dorsal horn tissues were extracted to detect the protein and mRNA expression of CX3CL1/CX3CR1 with Western blotting and RT-PCR seven days after modeling and after 20 days of intervention. The microglia morphology in spinal dorsal horn was observed with immunofluorescence after 20 days of intervention. Results:Seven days after modeling, compared with the normal group, the photothermal pain tolerance threshold increased in the model group and the sham group (P < 0.05); compared with the sham group, the cumulative pain score increased in the model group and Tuina group (P < 0.05). After ten days of intervention, the cumulative pain score was lower in Tuina group than in the model group (P < 0.05). After 20 days of intervention, both the photothermal pain tolerance threshold and cumulative pain score were lower in Tuina group than in the model group (P < 0.05). There was no significant difference in the expression of CX3CL1/CX3CR1 protein and mRNA on the seven days after modeling and after 20 days of intervention (P > 0.05). The microglia in the model group were partially activated or completely activated, while those in Tuina group were unactivated or partially activated after 20 days of intervention. Conclusion:Three Handing-Three Points can improve the pain function of sciatic nerve injured rats, which may associate with regulating microglia through the pathway other than CX3CL1/CX3CR1.

Biomechanical analysis of chemical acellular nerve allograft combined with bone marrow mesenchymal stem cell transplantation for repairing sciatic nerve injury / 中国组织工程研究

BACKGROUND: It is seldom reported that the effect of chemical acellular nerve allograft combined with bone marrow mesenchymal stem cell transplantation on sciatic nerve injury is evaluated by tensile mechanical properties. OBJECTIVE: To determine the effect of chemical acellular nerve allograft combined with bone marrow mesenchymal stem cell transplantation in the treatment of sciatic nerve injury. METHODS: Forty-five Sprague-Dawley rats were randomly divided into autologous nerve transplantation group, chemical acellular nerve allograft group, and chemical acellular nerve allograft and bone marrow mesenchymal stem cell transplantation group (n=15 per group). Models of 10-mm sciatic nerve injury were prepared. The injured sciatic nerves were repaired using autologous nerve, chemical acellular nerve allograft and chemical acellular nerve allograft and bone marrow mesenchymal stem cells. At 20 weeks after surgery, electrophysiological measurement and stretching experiments were carried out. RESULTS AND CONCLUSION: (1) The wave amplitude and motion conduction velocity were larger in the chemical acellular nerve allograft and bone marrow mesenchymal stem cell transplantation group and autologous nerve transplantation group than in the chemical acellular nerve allograft group (P < 0.05). (2) The tensile elastic limit strain, elastic limit stress, maximum strain and maximum stress were larger in the chemical acellular nerve allograft and bone marrow mesenchymal stem cell transplantation group and autologous nerve transplantation group than in the chemical acellular nerve allograft group (P < 0.05). (3) These results suggest that chemical acellular nerve allograft and bone marrow mesenchymal stem cell transplantation has obvious effect on repairing sciatic nerve injury.

Effects of electroacupuncture combined with nerve mobilization on lower limb motor function and mRNA and protein expression of C3 botoxin substrate 1 after sciatic nerve injury / 中华物理医学与康复杂志

Objective To investigate the effect of combining electroacupuncture with nerve mobilization to improve lower extremity motor function after sciatic nerve injury. And to document any changes in mRNA and protein expression of Ras-related C3 botoxin substrate 1. Methods 180 New Zealand rabbits were randomly divided into a normal control group, a model control group, an electroacupuncture group, a nerve mobilization group, and an elec-troacupuncture combined with nerve mobilization group, each of 36. Sciatic nerve injury was modelled using the clam-ping method in all except the normal control group. The control group had no intervention, while the nerve mobiliza-tion group, the electroacupuncture group and the combined group were treated with nerve mobilization, and/or elec-troacupuncture applied to the rabbit analogue of the jiaji acupoint. After 1, 2, and 4 weeks of treatment, toe reflex scores and modified Tarlov scores were used to assess any functional recovery. After 1, 2, and 4 weeks of treatment, 12 of the rabbits in each group were sacrificed and the sciatic nerve and the L4-L6 segments of the spinal cord were re-sected. The expression of Ras-related C3 botoxin substrate 1 mRNA and protein was detected using the polymerase chain reaction and western blotting. Results Sciatic nerve function and the expression of Ras-related C3 botoxin sub-strate 1 mRNA in the spinal cords and sciatic nerves of the three treatment groups were significantly higher than in the model control group at all three time points, but significantly lower than in the normal control group. The combined group′s results were significantly better than with electroacupuncture or nerve mobilization alone. After 1, 2, and 4 weeks of treatment, the average expression of Ras-related C3 botoxin substrate 1 protein in the spinal cords of the three treatment groups was significantly higher than the model control group′s average, but significantly lower than that of the normal control group at the same time point. After 1 week of treatment the average expression of Ras-related C3 botoxin substrate 1 protein in the spinal cords of the combined group was significantly higher than that in the group receiving electroacupuncture alone. After 2 and 4 weeks it was also significantly higher than the nerve mobilization group′s aver-age. After 1 week of treatment, the average expression of Ras-related C3 botoxin substrate 1 protein in the sciatic nerves of all three treatment groups was significantly lower than that of the control group. However, 1 and 3 weeks later the av-erage protein expression in the sciatic nerves was significantly higher than in the model control group, but significantly lower than in the normal control group at the same time points. The combined group′s average was then significantly higher than those of the groups receiving electroacupuncture or nerve mobilization alone at the same time point. Conclusion Nerve stimulation combined with electroacupuncture applied to the jiaji acupoint can promote the regener-ation of axons after sciatic nerve injury. The mechanism may be related to up-regulation of the Ras-related C3 botoxin substrate 1 gene and protein expression in the injured sciatic nerve and corresponding spinal cord segments.

Therapeutic Effect of Nerve Growth Factor Thermosensitive Gel on Sciatic Nerve Injury of Rats / 中国实验方剂学杂志

Objective: To prepare nerve growth factor(NGF) temperature sensitive in situ gel and investigate its therapeutic effect on sciatic nerve injury of rats.Method: NGF thermosensitive gel was prepared and its prescription was optimized by central composite design-response surface methodology.Fifty rats were randomly divided into the normal group,model group,NGF injection group(10 mg·L-1),NGF low-dose(10 mg·L-1) and high-dose(20 mg·L-1) thermosensitive gel groups,and sciatic nerve injury model of rats was established.The effect of NGF thermosensitive gel on the injury of sciatic nerve were comprehensively examined by taking rat behavior,sciatic nerve function index(SFI),time of withdrawal reflex,wet weight ratio of gastrocnemius muscle,and histomorphological changes as indicators.Result: The gelation temperature of NGF thermosensitive gel was 35.2℃ after the formulation being optimized,which was in line with the standard for injection.Four-eight weeks after operation,the SFI and wet weight ratio of gastrocnemius muscle in rats of NGF high-dose thermosensitive gel group were significantly higher than those in the model group and NGF injection group,but its time of withdrawal reflex was significantly lower than those in the model group and NGF injection group,and the effect was in a dose-dependent manner.Arrangement of regenerated nerve fibers in sciatic nerve injury area of rats from NGF high-dose thermosensitive gel group was more tidy,dense and continuous than that of the model group.Conclusion: NGF thermosensitive gel can promote repair of sciatic nerve injury in rats.

Effect of Tongma Qining Prescription on Sciatic Nerve Injury in Rats / 中国药学杂志

OBJECTIVE: To study the effect and mechanism of Tongma Qining prescription(TQP) on improving sciatic nerve injury (SNI) in rats. METHODS: The SNI model rats were replicated by using hemostatic forceps. The model rats were randomly divided into the model group, the mecobalamine group (mecobalamine 6×10-4 g•kg-1), the high-dose group (30 g •kg-1), the medium-dose group (15 g•kg-1) and the low-dose group (7.5 g•kg-1), and the sham operation group was set(10 mice per group). After continuous intragastric administration for 6 weeks, sciatic nerve index (SFI), sensory conduction velocity (SNCV) and hemodynamic indexes were detected in each group. HE staining were employed to detect the pathological alterations of sciatic nerve tissues. Then the expression of BDNF, NGF and VEGF in sciatic nerve were detected by IHC-P, the protein expression of NGF and VEGF were detected by Western blot. RESULTS: Tongma Qining prescription was able to alleviate sciatic nerve tissue injury in SNI rats, significantly improved SFI and SNCV (P<0.01), observably reduced the hematocrit, plasma viscosity and value of whole blood viscosity (high shearing and low sheating)(P<0.05), and markedly upregulated the expression level of BDNF, NGF, VEGF and the protein expression level of NGF, VEGF (P<0.01). CONCLUSION: The Tongma Qining prescription can significantly improve SNI in rats. Its action mechanism may be related to improving blood rheology, promoting angiogenesis and protecting nerve cells.

Effect of electroacupuncture on muscular atrophy and Slit/Robo signaling in sciatic nerve and lumbar spinal cord tissues in rats with sciatic nerve injury / 针刺研究

OBJECTIVE: To observe the effect of electroacupuncture (EA) of "Huantiao"(GB30) and" Zusanli"(ST36)on muscular atrophy and expression of Slit-Robo GTPase-activating protein(srGAP)1, 2 and 3 in the injured sciatic nerve and lumbar spinal cord tissues in sciatic nerve injury (SNI) rats, so as to reveal its mechanisms underlying improvement of peripheral nerve injury (PNI)．. METHODS: A total of 120 healthy male SD rats were randomly divided into control, sham-operation, model and EA groups (n＝30 rats in each) which were further divided into 7, 15 and 23 d subgroups (n＝10 rats in each subgroup). The SNI model was established by transecting the right sciatic nerve beneath the piriformis and immediately subsequent end-to-end suture. Rats of the sham operation group received an incision of the corresponding skin and suture. EA (5 Hz/20 Hz, 2－3 mA) was applied to the right GB30 and ST36 for 15 min, once daily, 6 days a week separately for 1，2 and 3 weeks. Rats in the sham-operation and model groups were grasped in the similar procedure as the EA group. The wet weight of gastrocnemius muscles (WWG) on both sides was measured to calculate the recovery rate (weight of the right WWG/weight of the left WWG×100%), and the expression levels of srGAP1, srGAP2 and srGAP3 proteins in the sciatic nerve and the spinal cord (L4-L6) tissues were detected by Western blot. RESULTS: After modeling and compared with the control and sham-operation groups, the recovery rate of WWG was significantly reduced, and the expression levels of srGAP1, srGAP2 and srGAP3 proteins of the sciatic nerve and lumbar spinal cord on day 7, 15 and 23 were considerably increased in the model group (P<0.01). Following the EA treatment, the reco-very rate of WWG was obviously increased and the expression levels of srGAP1, srGAP2 and srGAP3 proteins of both sciatic nerve and spinal cord on day 7, 15 and 23 were further significantly up-regulated in the EA group relevant to the model group (P<0.05，P<0.01). In addition, the expression levels of the 3 proteins in both sciatic nerve and lumbar spinal cord peaked on day 15 and attenuated on day 23. CONCLUSION: EA of GB30 and ST36 may relieve gastrocnemius atrophy in SNI rats, which is related to its function in up-regulating the Slit/Robo signaling in the sciatic nerve and lumbar spinal cord to promote the axonal targeting regeneration and repair of axonal plasma nutrition transportation.

Lower frequency electroacupuncture is better in promoting recovery of limb locomotion in rats with sciatic nerve injury by reducing local inflammatory reaction / 针刺研究

OBJECTIVE: To observe the effect of different frequencies (2 Hz, 100 Hz) of electroacupuncture (EA) on limb locomotion and the expression of inflammatory factors IL-1β, IL-6, TNF-α in sciatic nerve, and nuclear factor kappa B (NF-κB) in lumber(L)4-L5of spinal cord in rats with sciatic nerve injury (SNI), so as to reveal its mechanisms underlying improvement of SNI. METHODS: A total of 48 SD rats (half male and half female) were equally divided into blank control, model, low frequency (2 Hz) EA and high frequency (100 Hz) EA groups. The SNI model was established by clamping the spinal nerve. EA intervention (2 Hz, 100 Hz, 1 mA), starting on the 8th day after modeling, was applied to "Huantiao" (GB30) on the injured side for 15 min, once daily for 14 consecutive days. The sciatic function index (SFI) was calculated to assess the injured hindlimb recovery with reference to BAIN's and colleagues' methods. Histopathological changes of the sciatic nerve were displayed by H.E. staining. The expressions of IL-1β, IL-6 and TNF-α in the sciatic nerve tissue were detected by immunohistochemistry, and the expression of NF-κB in the spinal cord was detected by using Western blot. RESULTS: After modeling, the SFI level on day 8 was significantly decreased in the model group (P0.05). Following the treatment (at the 22nd day), the SFI values of both low frequency EA and high frequency EA groups were significantly increased (P<0.01), suggesting an improvement of the limb motor function, and the SFI of the low frequency EA group was notably higher than that of the high frequency EA group (P<0.01). In comparison with the blank control group, the expression levels of IL-1β, IL-6, TNF-α in the sciatic nerve and NF-κB protein in the spinal cord were significantly up-regulated (P<0.05). Following EA intervention, the increased expression levels of IL-1β, IL-6, TNF-α and NF-κB proteins were significantly down-regulated in both low frequency EA and high frequency EA groups (P<0.05), and the therapeutic effect of low frequency EA was markedly superior to that of high frequency EA in down-regulating the expression levels of IL-1β, IL-6, TNF-α and NF-κB protein (P<0.05). H.E. staining showed increase of Schwann cells in number, cellular swelling, and disintegration of the axons and myelin sheath, and appearance of vacuolar degeneration in the model group, which was relatively milder in both low frequency EA and high frequency EA groups, particularly in the low frequency EA group. CONCLUSION: EA of GB30 at 2 Hz and 100 Hz can promote the recovery of hindlimb motor function in SNI rats, which is probably related to its function in inhibiting the inflammatory response, and facilitating the repair of the damaged sciatic nerve. 2 Hz EA is better than 100 Hz EA in the therapeutic effect.

Effect of deep and shallow electroacupuncture stimulation at "Huantiao"(GB30) on expression of phosphorylated-p38 and phosphorylated-p53 proteins and apoptosis in dorsal root ganglia in sciatic nerve injury rats / 针刺研究

OBJECTIVE: To observe the effect of deep electroacupuncture (EA) stimulation at "Huantiao"(GB30) on hindlimb motor function and expression of p38 mitogen-activated protein kinase (p38 MAPK ) and p53 proteins in dorsal root ganglia (DRG) in rats with chronic constrictive injury (CCI) of sciatic nerve. METHODS: Forty-eight SD rats (half male and half female) were randomly divided into control, model, shallow EA (SEA) stimulation and deep EA (DEA) stimulation groups (n=12 in each group). The CCI model was constructed by implanting a silicone tube close to the sciatic nerve of the left hind limb. For DEA group and SEA group, filiform acupuncture needles were inserted into GB30 about 12-14 mm deep and 5-8 mm deep (monitored by using a high-frequency ultrasound device), respectively, followed by electrical stimulation (2 Hz/100 Hz, 1 mA) using an EA stimulator. The intervention was conducted for 15 min every time, once daily for 14 days. The sciatic nerve function index (SFI) calculated to assess the motor function status. Histopathological changes of the sciatic nerve were displayed by H.E. staining. The expression levels of phosphorylated-p38 MAPK (p-p38) and phosphorylated-tumor protein p53 (p-p53) in DRGs of L4-L5 on the affected side were observed by immunohistochemical staining. RESULTS: Following modeling, the SFI were significantly decreased (P<0.01), and the expression levels of p-p38 and p-p53 proteins of L4-L5 DRGs were considerably increased in the model group (P<0.05). After the intervention, the SFI were obviously increased, and the expression levels of p-p38 and p-p53 proteins notably down-regulated in both DEA and SEA groups relevant to the model group (P<0.01, P<0.05). The therapeutic effect of DEA was significantly superior to that of SEA in raising SFI and down-regulating expression le-vels of p-p38 and p-p53 proteins (P<0.01, P<0.05). H.E. staining showed disordered arrangement of the sciatic nerve fibers and myelin, disaggregation of the myelin and axons with deformity and vacuolation in some of them and with an increase of Schwann cells in the model group, which was relatively milder in both DEA and SEA groups. CONCLUSION: Both DEA and SEA at GB30 can obviously improve the motor function in CCI rats, which may be associated with its function in down-regulating the expression of p-p38 and p-p53 proteins in L4-L5 DRGs, restraining p38 MAPK signaling. The therapeutic effect of DEA is evidently better than that of SEA.

Effect of electroacupuncture at "Jiaji" (EX-B 2) points combined with nerve mobilization on motor function and mRNA and protein expression of RhoA in rabbits with sciatic nerve injury / 中国针灸

OBJECTIVE@#To observe the effects of electroacupuncture (EA) at "Jiaji" (EX-B 2) points combined with nerve mobilization on protein and mRNA expression of RhoA in rabbits with sciatic nerve injury, and to provide theoretical basis for the treatment of peripheral nerve injury by EA at "Jiaji" (EX-B 2) points combined with nerve mobilization.@*METHODS@#A total of 180 New Zealand rabbits were randomly divided into a normal control group, a model control group, a nerve mobilization group, an EA group, an EA plus nerve mobilization group, 36 rabbits in each group. Each group was further divided into a 1-week subgroup, 2-week subgroup and 4-week subgroup, 12 rabbits in each subgroup. The sciatic nerve injury model was made by clamping method. The rabbits in the normal control group did not receive any intervention. The rabbits in the model control group was normally fed after operation. The rabbits in the nerve mobilization group were treated with nerve mobilization; the manipulation lasted for 1 s and relaxed for 5 s, 10 times per day, 6 days per week. The rabbits in the EA group were treated with EA at "Jiaji" (EX-B 2) points (L-L), once a day, 30 min each time, 6 times per week. The rabbits in the EA plus nerve mobilization group were treated with EA at "Jiaji" (EX-B 2) points, followed by nerve mobilization. The function of sciatic nerve on the injured side was evaluated by toe tension reflex and modified Tarlov score; the tissues of corresponding segments of spinal cord L-L and sciatic nerve were taken; the expression of RhoA gene was detected by real-time PCR and the expression of RhoA protein was detected by Western Blot.@*RESULTS@#① Toe tension reflex and modified Tarlov score: at 1, 2 and 4 weeks, the scores in the model control group were lower than those in the normal control group (all 0.05); at 2 weeks, the expression in the nerve mobilization group was higher than that in the EA group (all <0.01); at 4 weeks, the expression in the nerve mobilization group was lower than that in the EA group (all <0.01).@*CONCLUSION@#The nerve mobilization and EA at "Jiaji" (EX-B 2) points could both promote the repair of injured sciatic nerve, which may be related to the down-regulation of RhoA expression, and the combination of the two methods has better effects.

Inhibition of KLF7-Targeting MicroRNA 146b Promotes Sciatic Nerve Regeneration / 神经科学通报·英文版

A previous study has indicated that Krüppel-like factor 7 (KLF7), a transcription factor that stimulates Schwann cell (SC) proliferation and axonal regeneration after peripheral nerve injury, is a promising therapeutic transcription factor in nerve injury. We aimed to identify whether inhibition of microRNA-146b (miR-146b) affected SC proliferation, migration, and myelinated axon regeneration following sciatic nerve injury by regulating its direct target KLF7. SCs were transfected with miRNA lentivirus, miRNA inhibitor lentivirus, or KLF7 siRNA lentivirus in vitro. The expression of miR146b and KLF7, as well as SC proliferation and migration, were subsequently evaluated. In vivo, an acellular nerve allograft (ANA) followed by injection of GFP control vector or a lentiviral vector encoding an miR-146b inhibitor was used to assess the repair potential in a model of sciatic nerve gap. miR-146b directly targeted KLF7 by binding to the 3'-UTR, suppressing KLF7. Up-regulation of miR-146b and KLF7 knockdown significantly reduced the proliferation and migration of SCs, whereas silencing miR-146b resulted in increased proliferation and migration. KLF7 protein was localized in SCs in which miR-146b was expressed in vivo. Similarly, 4 weeks after the ANA, anti-miR-146b increased KLF7 and its target gene nerve growth factor cascade, promoting axonal outgrowth. Closer analysis revealed improved nerve conduction and sciatic function index score, and enhanced expression of neurofilaments, P0 (anti-peripheral myelin), and myelinated axon regeneration. Our findings provide new insight into the regulation of KLF7 by miR-146b during peripheral nerve regeneration and suggest a potential therapeutic strategy for peripheral nerve injury.

Preparation of magnetic mNGF nanoparticles and treatment of rat sciatic nerve injury guided by outside magnetic field / 实用医学杂志

Objective To investigate the experimental effectofmagnetic mNGF nanoparticlesguided by out-side magnetic on the the sciatic nerve injury.Tianjin 300211,China.Methods As a drug carrier,PLGA connects magneticironparticles with and mNGF. The nanoparticle suspension liquid was prepared and its physicochemical properties was characterized. The model of the left sciatic nerve injury in rats was divided into 3 groups. Rats in Group A received intravenous tail vein injectionof magnetic mNGF nanoparticlesfollowed by two-hour outside mag-neticguidance once week.Group B received intravenous tail vein injection of magnetic mNGF nanoparticles without outside magnetic guidance once week. Group C received intravenous tail vein injection of the same content mNGF once week. Two months later,we measured the ischial nerve index(SFI)and madenerve electrical physiological ex-amination under anesthesia.We also measured quality ratio of triceps surae and did left sciatic nerve slice HE stain-ing.Finally,experimental data were statistically analyzed. Results The SFI,nerve conduction velocity and quality ratio of triceps surae of Group A were higher than those of Group B and Group B(P<0.05).There was no significant difference between Group B and Group C(P>0.05).The arrangement and density of the regeneration nerve in Group A were better than those of Group B and C under optical microscope. Conclusion The results showed that the magnetic mNGF nanoparticles guided by outside magnetic can promote the recovery of rat sciatic nerve injury.

Effects of Tuina of Three Handing-Three Points on Motor Function and Expression of NogoA and Its Receptor in Spinal Cord of Rats with Sciatic Nerve Injury / 中国康复理论与实践

@#Objective To investigate the effects of Tuina of Three Handing-Three Points on hindlimb motor function and expression of protein NogoA and its receptor NgR in the spinal cord of rats with sciatic nerve injury. Methods A total of 64 male Sprague-Dawley rats were randomly assigned to normal group (n = 16), sham group (n = 16), model group (n = 16) and Tuina group (n = 16). The model group and Tuina group prepared the sciatic nerve injury model with brace method. Tuina group received Tuina in methods of pressing, strumming and circular rubbing on Yinmen (BL37), Chengshan (BL57) and Yanglingquan (GB34) with Tuina manipulation emulator. The rats were assessed with oblique plate test seven days after operation and 20 days after Tuina. Then, the spinal cord tissues were extracted to detect the expression of protein of NogoA and NgR with Western blotting. Results The scores of the oblique plate test decreased in the model group and Tuina group seven days after operation compared with those in the normal group (P < 0.05), while the expression NgR increased (P < 0.05). The scores of the oblique plate test increased in Tuina group compared with those in the model group 20 days after Tuina (P < 0.05), while the expression of NgR decreased (P < 0.05), similar to the normal group (P > 0.05). Conclusion Tuina of Three Handing-Three Points can improve the hindlimb motor function in rats with sciatic nerve injury, which may be related to inhibition of the expression of NgR in the spinal cord after sciatic nerve injury.

Effects of One Finger Massage on Nerve Morphology and Function of Sciatic Nerve Injury Rats / 中国中医药信息杂志

Objective To study the effects of one finger massage on sciatic nerve injury rats. Methods The sciatic nerves were exposed, and the sciatic nerve was held by micro needles to make the sciatic nerve injury model. SD rats were randomly divided into sham-operation group, model group and massage group. In the sham-operation group, the sciatic nerves were exposed but not held. 7 d after the establishment of modeling, rats in massage group received one finger massage for 30 d. After 30 d, SFI and BBB of sciatic nerves were detected. HE staining, transmission electron microscope and immunochemistry assay were used to measure the changes of sciatic nerves. Results Compared with model group, massage group could speed up the recovery of SFI and increase BBB, promote the recovery of sciatic nerve morphology, increased protein level of S-100β, and enhance ultrastructure of newborn nerve growth and recovery. Conclusion One finger massage can effectively promote neurological and functional recovery after sciatic nerve injury in rats.

Effect of Cluster Needling of Scalp Acupuncture Combined with Treadmill Training on Axonal Regeneration and Expression of S-100 Protein in Rats with Sciatic Nerve Injury / 中国康复理论与实践

Objective To explore the effect of cluster needling of scalp acupuncture combined with treadmill training on axonal regener-ation and expression of S-100 protein in rats with sciatic nerve injury. Methods Ninety male Sprague-Dawley rats were randomly divided in-to blank group, sham group, model group, treadmill group and cluster needling of scalp point combined with treadmill group (combination group), each group was further divided into 7, 14 and 21 days subgroups, with 6 rats in each subgroup. The sciatic nerve of rats was clamped in the model group, the treadmill group and the combination group. The sham group was subjected to the same surgical procedure with sciat-ic nerve exposure but without crush injury. Three days after modeling, the treadmill group was treated with treadmill training, and the combi-nation group was treated with cluster needling of scalp acupuncture and treadmill training, while the other groups were grabbed regularly, with no other intervention. The sciatic functional index (SFI) was tested, the condition of axon growth was observed with HE staining, and the expression of S-100 protein was examined with immunohistochemistry at each time point. Results At each time point, SFI was higher, the axon grew better, and the S-100 protein was higher expressed in the treadmill group and the combination group than in the model group (P<0.05), especially in the combination group (P<0.05). Conclusion Cluster needling of scalp acupuncture combined with treadmill training can accelerate the regeneration of axons and increase the expression of S-100 protein and improve sciatic function after sciatic nerve injury.

Clinical Observations on Acupuncture Point Injection of Mouse Nerve Growth Factor for the Treatment of Sciatic Nerve Injury / 上海针灸杂志

ObjectiveTo investigate the clinical efficacy of points injection of mouse nerve growth factor in treating sciatic nerve injury.MethodAcupuncture point injection of injectable mouse nerve growth factorwas performed in 60 patients with sciatic nerve injury. Points Huantiao(GB30)and Zusanli(ST36)were selected. Treatment was given once daily, for a total of 30 times. Sensory and motor functions were assessed (MS and SS) and electromyography (EMG) was performed before and after treatment to compare motor nerve conduction velocities (MCV) and sensory nerve conduction velocities (SCV) in the injured nerves and EMG changes in the musclescontrolled by the injured nerves.ResultAfter treatment, nerve function was restored to more than S3M3, with an efficacy rate of 81.7%, in 49 patients. Neuro-electrophysiological study showed that regenerative potentials occurred, accounting for 71.7%,in43patients. Denervated potentials were fewer after treatment than before; there was a significant difference (P<0.05). The mean MCV value increased after treatment compared with before; there was a significant difference (P<0.05). ConclusionAcupuncture point injection of mouse nerve growth factor can markedly improve sensory and motor functions in the regions controlled by the injured sciatic nerve. It provides an effective way to promote the repair of sciatic nerves and the reconstruction of limb function after the nerves are injured.