RESUMO
BACKGROUND: The clinical efficacy of electroacupuncture in treating diabetic peripheral neuropathy (DPN) is significant, but the underlying mechanism of action is not clear. Considering that glucose-regulated protein 78 (GRP78) and caspase-12 are major proteins participating in cell apoptosis, we investigated the effects of "adjusting internal organs and dredging channel" electroacupuncture therapy on GRP78 and caspase-12 levels in streptozotocin (STZ)-diabetic rats to elucidate the mechanism of action. METHODS: Rats were first divided into two groups: one group was rendered diabetic with a single injection of 50 mg/kg STZ, whereas the other normal control group was injected with an equivalent volume of citrate buffer. The STZ-diabetic rats were randomly divided into three groups: model control and electroacupuncture- and mecobalamin-treated groups. After 12 weeks treatment, the therapeutic efficacy of electroacupuncture was assessed using sciatic nerves isolated from rats. In the electroacupuncture group, rats were treated by electroacupuncture for 20 minutes once daily for 6 days each week, with 1 day off, for 12 consecutive weeks. The selected acupressure points include bilateral acupressure points of BL13 (Fehu), BL20 (Pishu), BL23 (Shenshu), LI4 (Hegu), LR3 (faichong), ST36 (Zusanli), and SP6 (Sanyiniiao). Acupressure points were stimulated using a HuaTuo SDZ-V Electric Acupuncture Therapy Apparatus. The acupressure points of BL13 and BL23, as well as SP6 and LR3, were connected on the same side with a dilatational wave of 3 Hz (frequency ratio of 1 : 5) to stimulate the parts of the body to the extent that could be tolerated by the rat. As for the mecobalamin-treated groups, mecobalamin was administrated to rats intragastrically at a dose of 20 mg/kg once daily for 12 consecutive weeks. Immunofluorescence and western blot analysis were used to determine GRP78 and caspase-12 levels in sciatic nerves. In addition, cell apoptosis in sciatic nerves was determined using the terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end-labeling (TUNEL) assay. RESULTS: Electroacupuncture markedly reduced the pathological injury to sciatic nerves in STZ-diabetic rats. Moreover, electroacupuncture significantly downregulated GRP78 and caspase-12 and reduced cell apoptosis of sciatic nerves in DPN rats. CONCLUSIONS: Electroacupuncture improved DPN by downregulating GRP78 and caspase-12 and reducing cell apoptosis of sciatic nerves in STZ-diabetic rats, and further inhibited the occurrence of endoplasmic reticulum stress, thus preventing sciatic nerve injuries.
Assuntos
Caspase 12/metabolismo , Diabetes Mellitus Experimental/terapia , Neuropatias Diabéticas/prevenção & controle , Eletroacupuntura , Proteínas de Choque Térmico/metabolismo , Nervo Isquiático/enzimologia , Estreptozocina , Pontos de Acupuntura , Animais , Apoptose , Diabetes Mellitus Experimental/induzido quimicamente , Diabetes Mellitus Experimental/enzimologia , Diabetes Mellitus Experimental/patologia , Neuropatias Diabéticas/induzido quimicamente , Neuropatias Diabéticas/enzimologia , Neuropatias Diabéticas/patologia , Regulação para Baixo , Masculino , Ratos Sprague-Dawley , Nervo Isquiático/ultraestrutura , Transdução de SinaisRESUMO
The effects of low-level laser therapy (LLLT) and natural latex protein (F1, Hevea brasiliensis) were evaluated on crush-type injuries (15kg) to the sciatic nerve in the expressions of nerve growth factor (NGF) and vascular endothelium growth factor (VEGF) and ultrastructural morphology to associate with previous morphometric data using the same protocol of injury and treatment. Thirty-six male rats were allocated into six experimental groups (n = 6): 1-Control; 2-Exposed nerve; 3-Injured nerve; 4-LLLT (15J/cm2, 780nm, 30mW, Continuous Wave) treated injured nerve; 5-F1 (0,1mg) treated injured nerve; and 6-LLLT&F1 treated injured nerve. Four or eight weeks after, sciatic nerve samples were processed for analysis. NGF expression were higher (p<0.05) four weeks after in all injured groups in comparison to Control (Med:0.8; Q1:0; Q3:55.5%area). Among them, the Injured (Med:70.7; Q1:64.4; Q3:77.5%area) showed the highest expression, and F1 (Med:17.3; Q1:14.1; Q3:21.7%area) had the lowest. At week 8, NGF expressions decreased in the injured groups. VEGF was expressed in all groups; its higher expression was observed in the injured groups 4 weeks after (Injured. Med:29.5; F1. Med:17.7 and LLLT&F1. Med:19.4%area). At week 8, a general reduction of VEGF expression was noted, remaining higher in F1 (Med:35.1; Q1.30.6; Q3.39.6%area) and LLLT&F1 (Med:18.5; Q1:16; Q3:25%area). Ultrastructural morphology revealed improvements in the treated groups; 4 weeks after, the F1 group presented greater quantity and diameter of the nerve fibers uniformly distributed. Eight weeks after, the F1 and LLLT&F1 showed similar characteristics to the non-injured groups. In summary, these results and our previous studies indicated that F1 and LLLT may favorably influence the healing of nerve crush injury. Four weeks after nerve injury F1 group showed the best results suggesting recovery acceleration; at 8th week F1 and LLLT&F1 groups presented better features and higher vascularization that could be associated with VEGF maintenance.
Assuntos
Hevea/química , Terapia com Luz de Baixa Intensidade , Traumatismos dos Nervos Periféricos/terapia , Proteínas de Plantas/administração & dosagem , Nervo Isquiático/lesões , Animais , Lesões por Esmagamento/complicações , Modelos Animais de Doenças , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Látex/química , Masculino , Microscopia Eletrônica de Transmissão , Regeneração Nervosa/efeitos dos fármacos , Regeneração Nervosa/efeitos da radiação , Traumatismos dos Nervos Periféricos/etiologia , Proteínas de Plantas/isolamento & purificação , Ratos , Ratos Wistar , Recuperação de Função Fisiológica , Nervo Isquiático/fisiologia , Nervo Isquiático/ultraestrutura , Cicatrização/efeitos dos fármacos , Cicatrização/efeitos da radiaçãoRESUMO
Electrical stimulation could enhance nerve regeneration and functional recovery. The objective of this study was to evaluate the regenerative effects of implanted electrodes with different contacts in resected sciatic nerve. Sciatic nerve resection and microsurgical repair models were established and randomly divided into four groups (point contact, 1/4 circle contact; whole-circle contact; no electrodes as control). Electrical stimulation was performed and electrophysiological, morphological and histological exams (of the sciatic nerve and muscle) were conducted at 4 and 10 weeks post-implantation. Point and 1/4 circle contact groups showed significantly higher scores in the sciatic functional index (SFI), increased amplitude of compound muscle action potential (AMP) and motor nerve conduction velocity (MNCV) compared to the control group at both 4 and 10 weeks post-implantation. Point and 1/4 circle contact morphologically promoted sciatic nerve regeneration and reduced muscular atrophy with less mechanical injury to the nerve trunk observed compared with the whole-circle contact group at both 4 and 10 weeks post-implantation. Electrodes with point and 1/4 circle contacts represented an alternatively portable and effective method of electrical stimulation to facilitate injured sciatic nerve regeneration and reduce subsequent muscular atrophy, which might offer a promising approach for treating peripheral nerve injuries.
Assuntos
Terapia por Estimulação Elétrica/métodos , Eletrodos Implantados , Traumatismos dos Nervos Periféricos/terapia , Recuperação de Função Fisiológica , Nervo Isquiático/lesões , Animais , Masculino , Músculo Esquelético/inervação , Regeneração Nervosa/fisiologia , Traumatismos dos Nervos Periféricos/patologia , Ratos , Ratos Sprague-Dawley , Nervo Isquiático/patologia , Nervo Isquiático/ultraestruturaRESUMO
BACKGROUND: Electroacupuncture (EA) has been proved to be effective in treating certain neuropathic pain conditions. The mechanisms of pain relief by EA are not fully understood. There have been sporadic reports of damage in the peripheral nervous system (PNS) and regions of the central nervous system (CNS) at the ultrastructural level following peripheral nerve injury. However, information about possible systemic changes in the PNS and CNS after nerve injury is scarce. OBJECTIVES: The goal of this study was to examine the ultrastructural changes of the nervous system induced by a local injection of cobra venom into the sciatic nerve and to compare the ultrastructural changes in rats with or without treatment with EA or pregabalin. STUDY DESIGN: An experimental study. SETTING: Department of Anesthesiology, Pain Medicine, and Critical Care Medicine, Aviation General Hospital of China Medical University. METHODS: In this study, using an established model of sciatic neuralgia induced by local injection of cobra venom into the sciatic nerve, we examined ultrastructural changes of the PNS and CNS and how they respond to EA and pregabalin treatment. EA and pregabalin were given daily from postoperative day (POD) 14 to 36. Based on previous works, the frequency of EA stimulation of the ST36 and GB34 acupoints was held to 2/100 Hz variable. Pain sensitivity in the sciatic neuralgia rats with and without treatments was assessed using the von Frey test. Ultrastructural alterations were examined bilaterally in the prefrontal cortex, hippocampus, medulla oblongata; and the cervical, thoracic, and lumbar spinal cords on PODs 14, 40, and 60. Ultrastructural examinations were also carried out on the bilateral sciatic nerves and dorsal root ganglion (DRG) at the cervical, thoracic and lumbar levels. In rats treated with EA or pregabalin, the ultrastructure was examined on PODs 40 and 60. RESULTS: Behavioral signs of pain and systemic ultrastructural changes including demyelination were observed at all levels of the PNS and CNS in rats with sciatic neuralgia. After intervention, the mechanical withdrawal thresholds of the EA group and pregabalin group were significantly higher than that of the cobra venom group (P < 0.05). Both EA and pregabalin treatments partially reversed increased cutaneous sensitivity to mechanical stimulation. However, only the EA treatment was able to repair the ultrastructural damages caused by cobra venom. LIMITATIONS: The results confirm that peripheral nerve injury led to the ultrastructural damage at different levels of the CNS as demonstrated with electron microscopy; however, we need to further verify this at both the molecular level and in light microscope level. Sciatic neuralgia induced by cobra venom is a chemical injury, and whether this exactly mimics a peripheral nerve mechanical injury is still unclear. CONCLUSIONS: Local cobra venom injection leads to systemic neurotoxicity. EA and pregabalin alleviate pain via different mechanisms. KEY WORDS: Sciatic neuralgia, cobra venom, demyelination, electroacupuncture, pregabalin, rat model.
Assuntos
Eletroacupuntura/métodos , Neuralgia/patologia , Analgésicos/farmacologia , Animais , Encéfalo/patologia , Encéfalo/ultraestrutura , China , Venenos Elapídicos/toxicidade , Gânglios Espinais/patologia , Gânglios Espinais/ultraestrutura , Masculino , Neuralgia/induzido quimicamente , Limiar da Dor/efeitos dos fármacos , Limiar da Dor/fisiologia , Pregabalina/farmacologia , Ratos , Ratos Sprague-Dawley , Nervo Isquiático/efeitos dos fármacos , Nervo Isquiático/patologia , Nervo Isquiático/ultraestrutura , Medula Espinal/patologia , Medula Espinal/ultraestruturaRESUMO
OBJECTIVE: To investigate the effects and the underlying mechanisms of ShenFu Injection on paclitaxel-induced peripheral neuropathy. METHODS: Twenty-eight adult male Wister rats were randomized into 4 groups (n=7) : control group, paclitaxel group, paclitaxel combined with low or high dose of ShenFu Injection groups. Rats were intraperitoneally injected with paclitaxel 8 mg/kg every 4 d for a total of 4 doses except control group. From Day 1 of the experiment (injection),low dose (4 mL/kg) and high dose (8 mL/kg) of Shenfu Injection were intraperitoneally injected daily in the combination groups for a total of 21 d respectively,while normal saline (NS) was injected in control group in the same way instead. Mechanical withdraw threshold (MWT) and thermal withdraw latency (TWL) of rats' hind paw were measured before (0 d) and after the first injection (6 d,14 d). The level of nerve growth factor (NGF) in the serum was measured at 22 d before the euthanasia,and the ultrastructure of the sciatic nerve was observed with transmission electron microscope. RESULTS: The MWT and TWL of 14 d in paclitaxel group significantly increased compared with those of 0 d and control group ( P<0.05). The combination of paclitaxel with ShenFu Injection,especially the high dose ( P<0.05),significantly reduced the MWT and TWL when compared to paclitaxel group at 14 d. Compared with simultaneous control group,there was no remarkably increased MWT and TWL in the low and high dose of ShenFu Injection (P>0.05) . Compared with control group,the serum NGF level significantly decreased ( P<0.05) in paclitaxel group,while the serum NGF level in low and high dose of ShenFu Injection groups were higher than paclitaxel group,particularly in the high dose group ( P<0.05). When compared to control group,the sciatic nerve fiber structure in the paclitaxel group was generally damaged,including myelin sheath swelling,fragmentation and vacuolization,endoplasmic reticulum swelling and matrix structure disorder in Schwann cells. The structural damages were mitigated in the low dose and high dose groups,especially the latter one,when compared to the paclitaxel group. CONCLUSION: Shenfu Injection can reduce the peripheral neurotoxicity of paclitaxel by promoting the expression of NGF in serum.
Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Paclitaxel/efeitos adversos , Nervo Isquiático/efeitos dos fármacos , Animais , Masculino , Neurotoxinas/efeitos adversos , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Nervo Isquiático/ultraestruturaRESUMO
Herpes simplex virus type I (HSV-I) is a latent neuroinfection which can cause focal brain lesion. The role of HSV-infection in nerve regeneration has not been studied so far. The aim of the work was to study sciatic nerve regeneration in the presence of HSV-infection and the influence of an antiviral drug. BALB/c line mice were divided into five groups. Group 1 animals were infected with HSV-I. After resolution of neuroinfection manifestations the sciatic nerve of these animals was crushed. Group 2 mice were administered acyclovir following the same procedures. Groups 3-5 mice served as controls. Thirty days after the operation distal nerve stumps and m.gastrocnemius were studied morphologically and biochemically. Ultrastructural organization of the sciatic nerve in control animals remained intact. Morphometric parameters of the nerves from the experimental groups have not reach control values. However, in the group 1 diameter of nerve fibers was significantly smaller than in the group 2. Both nerve regeneration and m.gastrocnemius reinnervation were confirmed. The muscle hypotrophy was found in groups 1, 2, and 3 (the muscle fibers diameter decreased). Metabolic changes in the muscles of the infected animals (groups 1 and 2) were more pronounced than in control groups 3 and 4. The levels of TBA-active products, conjugated dienes, carbonyl and SH-groups were reduced in m.gastrocnemius of the experimental groups, however no significant difference associated with acyclovir administration was found. HSV-infection is not limited to the local neurodegenerative changes in the CNS but affects regeneration of the injured sciatic nerve. Anat Rec, 301:1734-1744, 2018. © 2018 Wiley Periodicals, Inc.
Assuntos
Aciclovir/uso terapêutico , Antivirais/uso terapêutico , Herpes Simples/tratamento farmacológico , Regeneração Nervosa/efeitos dos fármacos , Nervo Isquiático/efeitos dos fármacos , Aciclovir/farmacologia , Animais , Antivirais/farmacologia , Avaliação Pré-Clínica de Medicamentos , Camundongos Endogâmicos BALB C , Distribuição Aleatória , Nervo Isquiático/ultraestruturaRESUMO
AIM: Creatine is an endogenous molecule synthesized in the liver, kidney and pancreas from glycine and arginine and is important for mitochondrial metabolism. It is widely used as a supplement for improving muscle mass and function for many years. As it is expected to prevent apoptosis and diminish oxidative stress, it is also studied in a number of neurodegenerative diseases for its beneficial effect in recent years. We studied the effect of creatine on the peripheral nerve injury in an experimental rat crush injury model to obtain ultrastructural evidence. MATERIAL AND METHODS: Animals were randomly divided into 3 groups having 5 animals in each group. Group 1 was the control group, Group 2 the trauma group and Group 3 the trauma+creatine group. The first group served as sham control. In group 2 and group 3, sciatic nerves of the rats received crush injury using aneurysm clips. In group 3, daily 2 g/kg creatine monohydrate was administered via gavage after the trauma. Nerve samples were obtained at the 28th day after trauma for light and electron microscopic evaluation. RESULTS: Our comparative analysis results suggest a possible positive effect of creatine supplement on peripheral nerve regeneration as statistical analysis revealed significant differences between group 2 and group 3. Though our finding does not represent a miracle of regenerative support, beneficial usage of creatine is documented in the present study. CONCLUSION: Creatine supplement helps to diminish the harmful effects of peripheral nerve crush injury which is also supported by electron microscopy findings.
Assuntos
Creatina/uso terapêutico , Nervo Isquiático/ultraestrutura , Neuropatia Ciática/tratamento farmacológico , Neuropatia Ciática/patologia , Animais , Creatina/farmacologia , Suplementos Nutricionais , Masculino , Compressão Nervosa , Regeneração Nervosa/efeitos dos fármacos , Ratos , Ratos Wistar , Recuperação de Função Fisiológica/efeitos dos fármacos , Recuperação de Função Fisiológica/fisiologia , Nervo Isquiático/efeitos dos fármacos , Nervo Isquiático/lesões , Resultado do TratamentoRESUMO
BACKGROUND: Vitamin B12 and alpha lipoic acid (ALA) are known to promote functional and morphological recovery after peripheral nerve injury. OBJECTIVE: To compare the regenerative and neuroprotective effects of vitamin B12 and ALA treatment after sciatic nerve injury. METHODS: A total of 40 rats were randomly assigned to control (sciatic nerve exposure without injury or anastomosis), sham (sciatic nerve injury and epineural anastomosis were performed but no treatment was administered), PS (isotonic saline was administered for 12 weeks after surgery), ALA (2 mg/kg ALA was administered for 12 weeks after surgery), and vitamin B12 groups (2 mg/kg cyanocobalamin was administered for 12 weeks after surgery). Functional recovery was determined by footprint analysis, in vivo neurophysiology, and ex vivo histopathological examination. RESULTS: ALA treatment produced significant improvements in sciatic functional index values and non-significant improvements on electroneuromyography compared to vitamin B12 treatment. Upon histopathological examination, the regenerative effects of ALA were relevant to axonal structural recovery whereas vitamin B12 produced greater improvements in edema and myelination. CONCLUSIONS: While both vitamin B12 and ALA produced improvements after sciatic nerve injury, ALA was more functionally effective. The unique ultrastructural effects of vitamin B12 and ALA treatment should be considered in future studies.
Assuntos
Nervo Isquiático/efeitos dos fármacos , Ciática/tratamento farmacológico , Ácido Tióctico/uso terapêutico , Vitamina B 12/uso terapêutico , Complexo Vitamínico B/uso terapêutico , Animais , Avaliação Pré-Clínica de Medicamentos , Eletromiografia , Humanos , Masculino , Fármacos Neuroprotetores , Traumatismos dos Nervos Periféricos , Distribuição Aleatória , Ratos , Ratos Wistar , Recuperação de Função Fisiológica , Nervo Isquiático/ultraestrutura , Ácido Tióctico/farmacologia , Vitamina B 12/farmacologia , Complexo Vitamínico B/farmacologiaRESUMO
Oolong tea extract (OTE) was assessed for its potential as an electron stain to substitute for uranyl acetate (UA) in transmission electron microscopy (TEM). A comparative analysis of the ultrastructures of biological specimens (i.e. compound eye and sciatic nerve tissue) stained by different methods (UA and 0.1% OTE) has been performed. Results revealed that there was no significant difference between the OTE double staining method and the traditional double staining method, except the contrast in the OTE method was slightly lower than the UA method. Nevertheless, the OTE method yielded better or equal details in collagen fibres, neurofilaments and basal lamina in the mammalian sciatic nerve samples, as well as in the microvilli between the cornea and crystalline cone of the insect compound eye. On the whole, it was suggested that OTE could potentially be used as a nonradioactive and hazard-free alternative to UA in TEM staining.
Assuntos
Corantes/metabolismo , Microscopia Eletrônica de Transmissão/métodos , Compostos Organometálicos/metabolismo , Coloração e Rotulagem/métodos , Chá/metabolismo , Animais , Olho Composto de Artrópodes/ultraestrutura , Cães , Nervo Isquiático/ultraestrutura , TisanópterosRESUMO
OBJECTIVE: To observe the deregulation of autophagy in diabetic peripheral neuropathy (DPN) and investigate whether Jinmaitong ( JMT) alleviates DPN by inducing autophagy. METHODS: DPN models were established by streptozotocin-induced diabetic rats and Schwann cells (SCs) cultured in high glucose medium. The pathological morphology was observed by the improved Bielschowsky's nerve fiber axonal staining and the Luxol fast blue-neutral red myelin staining. The ultrastructure was observed by the transmission electron microscopy. Beclin1 level was detected by immunohistochemistry and Western blot. The proliferation of cultured SCs was detected by methylthiazolyldiphenyl-tetrazolium bromide. RESULTS: Diabetic peripheral nerve tissues demonstrated pathological morphology and reduced autophagic structure, accompanied with down-regulation of Beclin1. JMT apparently alleviated the pathological morphology change and increased the autophagy [in vivo, Beclin1 integral optical density (IOD) value of the control group 86.6±17.7, DM 43.9±8.8, JMT 73.3 ±17.8, P<0.01 or P<0.05, in vitro Beclin1 IOD value of the glucose group 0.47±0.25 vs the control group 0.88±0.29, P<0.05]. Consequently, inhibition of autophagy by 3-methyladenine resulted in a time- and concentration-dependent decrease of the proliferation of SCs (P<0.05, P<0.01). CONCLUSIONS: Down-regulation of autophagy in SCs might contribute to the pathogenesis of DPN. JMT alleviates diabetic peripheral nerve injury at least in part by inducing autophagy.
Assuntos
Autofagia , Diabetes Mellitus Experimental/complicações , Diabetes Mellitus Experimental/tratamento farmacológico , Neuropatias Diabéticas/complicações , Neuropatias Diabéticas/tratamento farmacológico , Medicamentos de Ervas Chinesas/uso terapêutico , Animais , Autofagia/efeitos dos fármacos , Axônios/efeitos dos fármacos , Axônios/patologia , Proteína Beclina-1/metabolismo , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Diabetes Mellitus Experimental/patologia , Neuropatias Diabéticas/patologia , Regulação para Baixo/efeitos dos fármacos , Medicamentos de Ervas Chinesas/farmacologia , Glucose/farmacologia , Imuno-Histoquímica , Masculino , Ratos Wistar , Células de Schwann/efeitos dos fármacos , Células de Schwann/patologia , Nervo Isquiático/efeitos dos fármacos , Nervo Isquiático/patologia , Nervo Isquiático/ultraestrutura , Coloração e RotulagemRESUMO
The objective of this study was to describe a perisciatic ultrasound-guided infiltration technique for treatment of deep gluteal syndrome and to report its preliminary clinical results. A mixture of saline (20 mL), a local anesthetic (4 mL), and a corticosteroid solution (1 mL) was infiltrated in the perisciatic region between the gluteus maximus and pelvitrochanteric muscles. Relative pain relief was achieved in 73.7% of the patients, with average preprocedural and postprocedural visual analog scale scores of 8.3 and 2.8, respectively. Fifty percent of patients reported recurrence of discomfort, and the average duration of the therapeutic effect in these patients was 5.3 weeks.
Assuntos
Anestesia Local/métodos , Anestésicos Locais/administração & dosagem , Síndrome do Músculo Piriforme/diagnóstico por imagem , Síndrome do Músculo Piriforme/tratamento farmacológico , Nervo Isquiático/ultraestrutura , Ciática/diagnóstico por imagem , Ciática/tratamento farmacológico , Ultrassonografia de Intervenção/métodos , Adulto , Feminino , Humanos , Masculino , Medição da Dor/efeitos dos fármacos , Projetos Piloto , Reprodutibilidade dos Testes , Nervo Isquiático/efeitos dos fármacos , Sensibilidade e Especificidade , Resultado do TratamentoRESUMO
We have used rat sciatic nerves submitted to freezing and freeze-fracture to determine the elemental composition of small domains of the peripheral nerve studied at high resolution by scanning electron microscopy. We found that myelin of Schwann cells is unique in its high content in phosphorus (P) that was more than 10 times higher than P measured in any other cells. This high concentration in P makes myelin chemistry suitable of monitoring at the subcellular level using the herein described methodology.
Assuntos
Bainha de Mielina/ultraestrutura , Fósforo/análise , Nervo Isquiático/ultraestrutura , Animais , Microanálise por Sonda Eletrônica , Feminino , Técnica de Fratura por Congelamento , Masculino , Microscopia Eletrônica de Varredura , Bainha de Mielina/metabolismo , Fósforo/metabolismo , Ratos , Ratos Wistar , Nervo Isquiático/metabolismoRESUMO
This study evaluated the effect of low-level laser therapy (LLLT; 15 J/cm(2)) and a latex protein (F1) on a crush injury of the sciatic (ischiadicus) nerve. Seventy-two rats (male, 250 g) were divided into 6 groups: CG, control; EG, exposed nerve; IG, injured nerve without treatment; LG, injured nerve with LLLT; HG, injured nerve with F1; and LHG, injured nerve with LLLT and F1. After 4 or 8 weeks, the animals were euthanized and samples of the sciatic nerve were collected for morphometric and high-resolution scanning electron microscopy (HRSEM) analysis. After 4 weeks, the morphometry revealed improvements in the treated animals, and the HG appeared to be the most similar to the CG; after 8 weeks, the injured groups showed improvements compared to the previous period, and the results of the treatment groups were more similar to one another. At HRSEM after 4 weeks, the treated groups were similar and showed improvement compared to the IG; after 8 weeks, the LHG and HG had the best results. In conclusion, the treatments resulted in improvement after the nerve injury, and this recovery was time-dependent. In addition, the use of the F1 resulted in the best morphometric and ultrastructural findings.
Assuntos
Hevea/química , Látex/administração & dosagem , Terapia com Luz de Baixa Intensidade/métodos , Microscopia Eletrônica de Varredura , Fitoterapia , Nervo Isquiático/lesões , Neuropatia Ciática/tratamento farmacológico , Animais , Látex/química , Masculino , Compressão Nervosa/efeitos adversos , Preparações de Plantas , Proteínas de Plantas/administração & dosagem , Proteínas de Plantas/química , Ratos , Ratos Wistar , Recuperação de Função Fisiológica , Nervo Isquiático/efeitos dos fármacos , Nervo Isquiático/efeitos da radiação , Nervo Isquiático/ultraestrutura , Neuropatia Ciática/patologia , Neuropatia Ciática/radioterapia , Fatores de TempoRESUMO
Regeneration and functional reconstruction of peripheral nerve defects remained a significant clinical challenge. Nerve guide conduits, with seed cells or neurotrophic factors (NTFs), had been widely used to improve the repair and regeneration of injured peripheral nerve. Pyrroloquinoline quinone (PQQ) was an antioxidant that can stimulate nerve growth factors (NGFs) synthesis and accelerate the Schwann cells (SCs) proliferation and growth. In present study, three kinds of nerve guide conduits were constructed: one from cellulose/SPI hollow tube (CSC), another from CSC combined with SCs (CSSC), and the third one from CSSC combined with PQQ (CSSPC), respectively. And then they were applied to bridge and repair the sciatic nerve defect in rats, using autograft as control. Effects of different nerve guide conduits on the nerve regeneration were comparatively evaluated by general analysis, sciatic function index (SFI) and histological analysis (HE and TEM). Newly-formed regenerative nerve fibers were observed and running through the transparent nerve guide conduits 12 weeks after surgery. SFI results indicated that the reconstruction of motor function in CSSPC group was better than that in CSSC and CSC groups. HE images from the cross-sections and longitudinal-sections of the harvested regenerative nerve indicated that regenerative nerve fibers had been formed and accompanied with new blood vessels and matrix materials in the conduits. TEM images also showed that lots of fresh myelinated and non-myelinated nerve fibers had been formed. Parts of vacuolar, swollen and abnormal axons occurred in CSC and CSSC groups, while the vacuolization and swell of axons was the least serious in CSSPC group. These results indicated that CSSPC group had the most ability to repair and reconstruct the nerve structure and functions due to the comprehensive contributions from hollow CSC tube, SCs and PQQ. As a result, the CSSPC may have the potential for the applications as nerve guide conduits in the field of nerve tissue engineering.
Assuntos
Antioxidantes/uso terapêutico , Celulose/química , Regeneração Nervosa/efeitos dos fármacos , Cofator PQQ/uso terapêutico , Células de Schwann/citologia , Nervo Isquiático/fisiologia , Proteínas de Soja/química , Alicerces Teciduais/química , Animais , Antioxidantes/administração & dosagem , Regeneração Tecidual Guiada , Masculino , Cofator PQQ/administração & dosagem , Ratos , Ratos Sprague-Dawley , Células de Schwann/efeitos dos fármacos , Nervo Isquiático/lesões , Nervo Isquiático/patologia , Nervo Isquiático/ultraestrutura , Engenharia TecidualRESUMO
This study aimed to investigate whether or not ginkgo biloba extract (EGb 761) enhances peripheral nerve regeneration and vascularization after repair using acellular nerve allografts (ANA). Seventy-two Sprague-Dawley rats were randomly divided into three experimental groups: a unilateral 15-mm sciatic nerve defect was created and repaired with an autologous graft (autograft group); the same defect was repaired with an 18 mm ANA with an i.p. injection of normal saline for 10 days (saline group); and in the final group, the same defect was repaired with an 18 mm ANA with an i.p. injection of EGb 761 for 10 days (EGb 761 group). Axon outgrowth and vascularization were evaluated by immunocytochemistry 14 days post-implantation. The expression of genes associated with angiogenesis was analyzed by real-time polymerase chain reaction (PCR) seven days post-implantation. Compared with the saline group, rats in the EGb 761 group significantly increased the number of myelinated fibers and the average diameter of the nerves within the graft. There is no significant difference between the EGb 761 group and the autograft group. The expression of CD34 and NF200 was significantly higher in the EGb 761 group than in the saline group. Additionally, EGb 761 treatment increased the expression of several angiogenesis-related genes, including Vegf, SOX18, Prom 1, and IL-6. In conclusion, ANA repair with EGb 761 treatment demonstrates effects on peripheral nerve regeneration and vascularization that are equal to those of autologous graft repair, and that are superior to ANA repair alone.
Assuntos
Aloenxertos/irrigação sanguínea , Neovascularização Fisiológica/efeitos dos fármacos , Regeneração Nervosa/efeitos dos fármacos , Extratos Vegetais/farmacologia , Nervo Isquiático/transplante , Aloenxertos/efeitos dos fármacos , Animais , Antígenos CD34/metabolismo , Axônios/efeitos dos fármacos , Modelos Animais de Doenças , Ginkgo biloba , Imuno-Histoquímica , Masculino , Músculos/efeitos dos fármacos , Bainha de Mielina/efeitos dos fármacos , Bainha de Mielina/ultraestrutura , Tamanho do Órgão/efeitos dos fármacos , Ratos Sprague-Dawley , Nervo Isquiático/efeitos dos fármacos , Nervo Isquiático/patologia , Nervo Isquiático/ultraestruturaRESUMO
Local effect of acetyl salicylic acid (ASA) on peripheral nerve regeneration was studied using a rat sciatic nerve transection model. Forty-five male healthy White Wistar rats were divided into three experimental groups (n = 15), randomly: Sham-operation (SHAM), control (SIL), and ASA-treated (SIL/ASA) groups. In SHAM group after anesthesia left sciatic nerve was exposed through a gluteal muscle incision and after homeostasis the muscle was sutured. In SIL group the left sciatic nerve was exposed the same way and transected proximal to tibio-peroneal bifurcation leaving a 10-mm gap. Proximal and distal stumps were each inserted into a silicone tube and filled with 10 µl phosphate buffered solution. In SIL/ASA group defect was bridged using a silicone tube filled with 10 µl acetyl salisylic acid (0.1 mg/ml). Each group was subdivided into three subgroups of five animals each and were studied 4, 8, and 12 weeks after surgery. Data were analyzed statistically by factorial analysis of variance (ANOVA) and the Bonferroni test for pair-wise comparisons. Functional study confirmed faster and better recovery of regenerated axons in SIL/ASA than in SIL group (p < 0.05). Gastrocnemius muscle mass in SIL/ASA was significantly more than in SIL group. Morphometric indices of regenerated fibers showed that the number and diameter of the myelinated fibers in SIL/ASA were significantly higher than in control group. In immuohistochemistry, location of reactions to S-100 in SIL/ASA was clearly more positive than in SIL group. Response to local treatment of ASA demonstrates that it influences and improves functional recovery of peripheral nerve regeneration.
Assuntos
Aspirina/uso terapêutico , Regeneração Nervosa/efeitos dos fármacos , Traumatismos dos Nervos Periféricos/tratamento farmacológico , Nervo Isquiático/lesões , Administração Tópica , Animais , Aspirina/administração & dosagem , Axotomia , Avaliação Pré-Clínica de Medicamentos , Regeneração Tecidual Guiada/instrumentação , Regeneração Tecidual Guiada/métodos , Masculino , Microcirurgia , Músculo Esquelético/inervação , Atrofia Muscular/prevenção & controle , Fibras Nervosas Mielinizadas/efeitos dos fármacos , Fibras Nervosas Mielinizadas/ultraestrutura , Traumatismos dos Nervos Periféricos/patologia , Complicações Pós-Operatórias/prevenção & controle , Distribuição Aleatória , Ratos , Ratos Wistar , Recuperação de Função Fisiológica , Nervo Isquiático/cirurgia , Nervo Isquiático/ultraestrutura , Alicerces Teciduais , CaminhadaRESUMO
Earlier research has demonstrated that hyperbaric oxygen (HBO2) can produce an antinociceptive effect in models of acute pain. Recent studies have revealed that HBO2 can produce pain relief in animal models of chronic pain as well. The purpose of the present investigation was to ascertain whether HBO2 treatment might suppress allodynia in rats with neuropathic pain and whether this effect might be blocked by the opioid antagonist naltrexone (NTX). Male Sprague Dawley rats were subjected to a sciatic nerve crush under anesthesia and mechanical thresholds were assessed using an electronic von Frey anesthesiometer. The time course of the HBO2-induced anti-allodynic effect in different treatment groups was plotted, and the area-under-the-curve (AUC) was determined for each group. Seven days after the nerve crush procedure, rats were treated with HBO2 at 3.5 atm absolute (ATA) for 60 min and exhibited an anti-allodynic effect, compared to nerve crush-only control rats. Twenty-four hours before HBO2 treatment, another group of rats was implanted with Alzet(®) osmotic minipumps that continuously released NTX into the lateral cerebral ventricle for 7 days. These NTX-infused, HBO2-treated rats exhibited an allodynic response comparable to that exhibited by rats receiving nerve crush only. Analysis of the AUC data showed that HBO2 significantly reduced the nerve crush-induced allodynia; this anti-allodynic effect of HBO2 was reversed by NTX. These results implicate opioid receptors in the pain relief induced by HBO2.
Assuntos
Hiperalgesia/terapia , Oxigenoterapia Hiperbárica , Neuralgia/metabolismo , Receptores Opioides/metabolismo , Nervo Isquiático/fisiopatologia , Medula Espinal/fisiopatologia , Animais , Oxigenoterapia Hiperbárica/métodos , Masculino , Naltrexona/administração & dosagem , Naltrexona/uso terapêutico , Antagonistas de Entorpecentes/uso terapêutico , Neuralgia/tratamento farmacológico , Medição da Dor/métodos , Ratos , Ratos Sprague-Dawley , Nervo Isquiático/efeitos dos fármacos , Nervo Isquiático/lesões , Nervo Isquiático/ultraestrutura , Medula Espinal/efeitos dos fármacosRESUMO
This study analyzed the effects of a low-level laser therapy (LLLT, 15 J/cm(2), 780 nm wavelength) and the natural latex protein (P1, 0.1%) in sciatic nerve after crush injury (15 Kgf, axonotmesis) in rats. Sixty rats (male, 250 g) were allocated into the 6 groups (n = 10): CG-control group; EG-nerve exposed; IG-injured nerve without treatment; LG-crushed nerve treated with LLLT; PG-injured nerve treated with P1; and LPG-injured nerve treated with LLLT and P1. After 4 or 8 weeks, the nerve samples were processed for morphological, histological quantification and ultrastructural analysis. After 4 weeks, the myelin density and morphological characteristics improved in groups LG, PG, and LPG compared to IG. After 8 weeks, PG, and LPG were similar to CG and the capillary density was higher in the LG, PG, and LPG. In the ultrastructural analysis the PG and LPG had characteristics that were similar to the CG. The application of LLLT and/or P1 improved the recovery from the nerve crush injury, and in the long term, the P1 protein was the better treatment used, since only the application of LLLT has not reached the same results, and these treatments applied together did not potentiate the recovery.
Assuntos
Terapia com Luz de Baixa Intensidade , Nervo Isquiático/efeitos dos fármacos , Nervo Isquiático/lesões , Animais , Hevea/química , Látex/administração & dosagem , Látex/química , Masculino , Compressão Nervosa , Proteínas de Plantas/administração & dosagem , Proteínas de Plantas/química , Proteínas de Plantas/isolamento & purificação , Ratos , Ratos Wistar , Nervo Isquiático/efeitos da radiação , Nervo Isquiático/ultraestruturaRESUMO
BACKGROUND: Electroacupuncture (EA) is effective in relieving pain in patients with postherpetic neuralgia (PHN). However, the mechanism underlying the therapeutic effect of EA in PHN is still unclear. Systemic injection of resiniferatoxin (RTX), an ultrapotent analog of TRPV1 agonist, in adult rats can reproduce the clinical symptoms of PHN by ablating TRPV1-expressing sensory neurons. In this study, we determined the beneficial effect of EA and the potential mechanisms in this rat model of PHN. METHODS: PHN was induced in rats by a single injection of RTX. Thermal hyperalgesia was tested with a radiant heat stimulus, and mechanical allodynia was quantified with von Frey filaments. TRPV1 receptors were shown by using immunofluorescence labeling. The ultrastructural changes of the sciatic nerve were assessed by electron microscopic examination. The sprouting of myelinated primary afferent terminals into the spinal dorsal horn was mapped by using the transganglionic tracer cholera toxin B-subunit (CTB). RESULTS: RTX injection diminished thermal sensitivity and gradually induced tactile allodynia within 3 weeks. EA applied to GB30 and GB34 at 2 and 15 Hz, but not 100 Hz, significantly increased the thermal sensitivity 4 weeks after treatment and decreased the tactile allodynia 2 weeks after treatment in RTX-treated rats. EA treatment at 2 and 15 Hz recovered the loss of TRPV1-positive dorsal root ganglion neurons and their central terminals of afferent fibers in the spinal superficial dorsal horn of RTX-treated rats. Moreover, EA significantly reduced the loss of unmyelinated fibers and the damage of the myelinated nerve fibers of RTX-treated rats. Furthermore, EA at 2 and 15 Hz inhibited the sprouting of myelinated primary afferent terminals into the spinal lamina II of RTX-treated rats. CONCLUSIONS: EA treatment improves thermal perception by recovering TRPV1-positive sensory neurons and nerve terminals damaged by RTX. EA Also reduces RTX-induced tactile allodynia by attenuating the damage of myelinated afferent nerves and their abnormal sprouting into the spinal lamina II. Our study provides new information about the mechanisms of the therapeutic actions of EA in the treatment of PHN.
Assuntos
Eletroacupuntura , Hiperalgesia/patologia , Hiperalgesia/terapia , Neuralgia Pós-Herpética/patologia , Neuralgia Pós-Herpética/terapia , Temperatura , Animais , Toxina da Cólera/metabolismo , Modelos Animais de Doenças , Diterpenos , Gânglios Espinais/metabolismo , Gânglios Espinais/patologia , Masculino , Bainha de Mielina/metabolismo , Neurônios Aferentes/metabolismo , Neurônios Aferentes/patologia , Células do Corno Posterior/metabolismo , Células do Corno Posterior/patologia , Subunidades Proteicas/metabolismo , Ratos , Ratos Sprague-Dawley , Nervo Isquiático/patologia , Nervo Isquiático/ultraestrutura , Canais de Cátion TRPV/metabolismo , Fatores de TempoRESUMO
OBJECTIVE: To study the effects of the Chinese medicine Jinmaitong Capsule (, JMT) on the pathomorphology of sciatic nerves, ciliary neurotrophic factor (CNTF), and the mRNA expressions of CNTF in rats with streptozotocin-induced diabetes mellitus (STZ-DM). METHODS: The animal model was established by one time intraperitoneal injection of streptozotocin. The rats were simply divided by random into 5 groups including model group, low-dose JMT group (JL), medium-dose JMT group (JM), high-dose JMT group (JH) and neurotropin group. For each of the above 5 groups, a group of 10 normal Wistar rats matched in body weight, age and gender were set as normal group. Intragastric administrations were started after the animal model established. The JL group were administered with five times the JMT dose recommended for a human adult; the JM group were administered with ten times the JMT dose recommended for a human adult; the JH group were administered with twenty times the JMT dose recommended for a human adult. The neurotropin group was administered with ten times the neurotropin dose recommended for a human adult. All rats were given intragastric administration for 16 weeks and then killed. In the 4th, 8th, 12th, 16th week, body weight and blood glucose level were detected before and after the intervention. The morphologic changes of the sciatic nerves were observed by optical microscope and transmission electron microscope. The CNTFmRNA expressions were detected by real-time fluorescent quantitative polymerase chain protein, and the CNTF protein expressions were detected by immunohistochemical method. RESULTS: The blood glucose levels of the STZ-DM rats were much higher than normal group (P<0.01), and there was no apparent difference between any treatment groups and the model group (P>0.05). Before and after the intervention in the 4th, 8th, 12th, 16th week, there were no significant differences in the body weight among all the groups (P>0.05). The sciatic nerves of STZ-DM rats might have pathomorphological changes in axons, myelin sheaths, and interstitium. The levels of CNTF and CNTF-mRNA expressions in the STZ-DM rats were both significantly decreased (P<0.01). The sciatic nerves of STZ-DM rats might have pathomorphological changes in axons, myelin sheaths, and interstitium. CONCLUSION: JMT could improve the pathomorphology of sciatic nerves by increasing CNTF's and CNTF-mRNA expressions in sciatic nerve tissues, and promote the repair and regeneration of damaged nerve fibers.