Concomitant treatment of ureteral calculi and ipsilateral pelvic sciatic nerve schwannoma with transperitoneal laparoscopic approach: A case report

BACKGROUND Schwannomas are rare peripheral neural myelin sheath tumors that originate from Schwann cells.Of the different types of schwannomas,pelvic sciatic nerve schwannoma is extremely rare.Definite preoperative diagnosis of pelvic schwannomas is difficult,and surgical resection is the gold standard for its definite diagnosis and treatment.CASE SUMMARY We present a case of pelvic schwannoma arising from the sciatic nerve that was detected in a 40-year-old man who underwent computed tomography for intermittent right lower back pain caused exclusively by a right ureteral calculus.Subsequently,successful transperitoneal laparoscopic surgery was performed for the intact removal of the stone and en bloc resection of the schwannoma.The total operative time was 125 min,and the estimated blood loss was inconspicuous.The surgical procedure was uneventful.The patient was discharged on postoperative day 5 with the simultaneous removal of the urinary catheter.However,the patient presented with motor and sensory disorders of the right lower limb,caused by partial damage to the right sciatic nerve.No tumor recurrence was observed at the postoperative appointment.CONCLUSION Histopathological examination of the specimen confirmed the diagnosis of a schwannoma.Thus,laparoscopic surgery is safe and feasible for concomitant extirpation of pelvic schwannomas and other pelvic and abdominal diseases that require surgical treatment.

Silk-based nerve guidance conduits with macroscopic holes modulate the vascularization of regenerating rat sciatic nerve

Peripheral nerve injuries induce a severe motor and sensory deficit. Since the availability of autologous nerve transplants for nerve repair is very limited, alternative treatment strategies are sought, including the use of tubular nerve guidance conduits(tNGCs). However, the use of tNGCs results in poor functional recovery and central necrosis of the regenerating tissue, which limits their application to short nerve lesion defects(typically shorter than 3 cm). Given the importance of vascularization in nerve regeneration, we hypothesized that enabling the growth of blood vessels from the surrounding tissue into the regenerating nerve within the tNGC would help eliminate necrotic processes and lead to improved regeneration. In this study, we reported the application of macroscopic holes into the tubular walls of silk-based tNGCs and compared the various features of these improved silk^(+) tNGCs with the tubes without holes(silk^(–) tNGCs) and autologous nerve transplants in an 8-mm sciatic nerve defect in rats. Using a combination of micro-computed tomography and histological analyses, we were able to prove that the use of silk^(+) tNGCs induced the growth of blood vessels from the adjacent tissue to the intraluminal neovascular formation. A significantly higher number of blood vessels in the silk^(+) group was found compared with autologous nerve transplants and silk^(–), accompanied by improved axon regeneration at the distal coaptation point compared with the silk^(–) tNGCs at 7 weeks postoperatively. In the 15-mm(critical size) sciatic nerve defect model, we again observed a distinct ingrowth of blood vessels through the tubular walls of silk^(+) tNGCs, but without improved functional recovery at 12 weeks postoperatively. Our data proves that macroporous tNGCs increase the vascular supply of regenerating nerves and facilitate improved axonal regeneration in a short-defect model but not in a critical-size defect model. This study suggests that further optimization of the macroscopic holes silk^(+) tNGC approach containing macroscopic holes might result in improved grafting technology suitable for future clinical use.

Polyethylene glycol has immunoprotective effects on sciatic allografts, but behavioral recovery and graft tolerance require neurorrhaphy and axonal fusion

Behavioral recovery using(viable)peripheral nerve allografts to repair ablation-type(segmental-loss)peripheral nerve injuries is delayed or poor due to slow and inaccurate axonal regeneration.Furthermore,such peripheral nerve allografts undergo immunological rejection by the host immune system.In contrast,peripheral nerve injuries repaired by polyethylene glycol fusion of peripheral nerve allografts exhibit excellent behavioral recovery within weeks,reduced immune responses,and many axons do not undergo Wallerian degeneration.The relative contribution of neurorrhaphy and polyethylene glycol-fusion of axons versus the effects of polyethylene glycol per se was unknown prior to this study.We hypothesized that polyethylene glycol might have some immune-protective effects,but polyethylene glycol-fusion was necessary to prevent Wallerian degeneration and functional/behavioral recovery.We examined how polyethylene glycol solutions per se affect functional and behavioral recovery and peripheral nerve allograft morphological and immunological responses in the absence of polyethylene glycol-induced axonal fusion.Ablation-type sciatic nerve injuries in outbred Sprague–Dawley rats were repaired according to a modified protocol using the same solutions as polyethylene glycol-fused peripheral nerve allografts,but peripheral nerve allografts were loose-sutured(loose-sutured polyethylene glycol)with an intentional gap of 1–2 mm to prevent fusion by polyethylene glycol of peripheral nerve allograft axons with host axons.Similar to negative control peripheral nerve allografts not treated by polyethylene glycol and in contrast to polyethylene glycol-fused peripheral nerve allografts,animals with loose-sutured polyethylene glycol peripheral nerve allografts exhibited Wallerian degeneration for all axons and myelin degeneration by 7 days postoperatively and did not recover sciatic-mediated behavioral functions by 42 days postoperatively.Other morphological signs of rejection,such as collapsed Schwann cell basal lamina tubes,were absent in polyethylene glycol-fused peripheral nerve allografts but commonly observed in negative control and loose-sutured polyethylene glycol peripheral nerve allografts at 21 days postoperatively.Loose-sutured polyethylene glycol peripheral nerve allografts had more pro-inflammatory and less anti-inflammatory macrophages than negative control peripheral nerve allografts.While T cell counts were similarly high in loose-sutured-polyethylene glycol and negative control peripheral nerve allografts,loose-sutured polyethylene glycol peripheral nerve allografts expressed some cytokines/chemokines important for T cell activation at much lower levels at 14 days postoperatively.MHCI expression was elevated in loose-sutured polyethylene glycol peripheral nerve allografts,but MHCII expression was modestly lower compared to negative control at 21 days postoperatively.We conclude that,while polyethylene glycol per se reduces some immune responses of peripheral nerve allografts,successful polyethylene glycol-fusion repair of some axons is necessary to prevent Wallerian degeneration of those axons and immune rejection of peripheral nerve allografts,and produce recovery of sensory/motor functions and voluntary behaviors.Translation of polyethylene glycol-fusion technologies would produce a paradigm shift from the current clinical practice of waiting days to months to repair ablation peripheral nerve injuries.

A hyaluronic acid granular hydrogel nerve guidance conduit promotes regeneration and functional recovery of injured sciatic nerves in rats

A hyaluronic acid granular hydrogel can promote neuronal and astrocyte colony formation and axonal extension in vitro,suggesting that the hydrogel can simulate an extracellular matrix structure to promote neural regeneration.However,in vivo experiments have not been conducted.In this study,we transplanted a hyaluronic acid granular hydrogel nerve guidance conduit to repair a 10-mm long sciatic nerve gap.The Basso,Beattie,and Bresnahan locomotor rating scale,sciatic nerve compound muscle action potential recording,Fluoro-Gold retrograde tracing,growth related protein 43/S100 immunofluorescence staining,transmission electron microscopy,gastrocnemius muscle dry/wet weight ratio,and Masson’s trichrome staining results showed that the nerve guidance conduit exhibited similar regeneration of sciatic nerve axons and myelin sheath,and recovery of the electrophysiological function and motor function as autologous nerve transplantation.The conduit results were superior to those of a bulk hydrogel or silicone tube transplant.These findings suggest that tissue-engineered nerve conduits containing hyaluronic acid granular hydrogels effectively promote the morphological and functional recovery of the injured sciatic nerve.The nerve conduits have the potential as a material for repairing peripheral nerve defects.

Platelet-rich plasma promotes peripheral nerve regeneration after sciatic nerve injury

The effect of platelet-rich plasma on nerve regeneration remains controversial.In this study,we established a rabbit model of sciatic nerve small-gap defects with preserved epineurium and then filled the gaps with platelet-rich plasma.Twenty-eight rabbits were divided into the following groups(7 rabbits/group):model,low-concentrati on PRP(2.5-3.5-fold concentration of whole blood platelets),medium-concentration PRP(4.5-6.5-fold concentration of whole blood platelets),and high-concentration PRP(7.5-8.5-fold concentration of whole blood platelets).Electrophysiological and histomorphometrical assessments and proteomics analysis we re used to evaluate regeneration of the sciatic nerve.Our results showed that platelet-rich plasma containing 4.5-6.5-and 7.5-8.5-fold concentrations of whole blood platelets promoted repair of sciatic nerve injury.Proteomics analysis was performed to investigate the possible mechanism by which platelet-rich plasma promoted nerve regeneration.Proteomics analysis showed that after sciatic nerve injury,platelet-rich plasma increased the expression of integrin subunitβ-8(ITGB8),which participates in angiogenesis,and differentially expressed proteins were mainly enriched in focal adhesion pathways.Additionally,two key proteins,ribosomal protein S27 a(RSP27 a)and ubiquilin 1(UBQLN1),which were selected after protein-protein interaction analysis,are involved in the regulation of ubiquitin levels in vivo.These data suggest that platelet-rich plasma promotes peripheral nerve regeneration after sciatic nerve injury by affecting angiogenesis and intracellular ubiquitin levels.

Comparison of the Nerve Regeneration Capacity and Characteristics between Sciatic Nerve Crush and Transection Injury Models in Rats

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.

Label-free quantitative proteomics analysis models in vivo and in vitro reveal key proteins and potential roles in sciatic nerve injury

Background:The underlying mechanism of sciatic nerve injury(SNI)is a common motor functional disorder,necessitates further research.Methods:A rat model of SNI was established,with the injury group subjected to compressive injury of the right sciatic nerve exposed at the midpoint of the thigh and the sham surgery group undergoing the same surgical procedure.An oxygen-glucose deprivation model was employed to simulate in vitro SNI in PC12 cells.Following data acquisition and quality control,differentially expressed proteins(DEPs)in each model were identified through differential analysis,and enrichment analysis was used to explore the potential functions and pathways of the DEPs.Venn diagrams were drawn,and DEPs from both in vivo and in vitro SNI models were imported into the STRING database to construct a protein-protein interaction network and screen for hub proteins.Results:After the peptide segments obtained from rat nerve blockade and PC12 cells met quality requirements,258 DEPs were identified in rat nerve samples,and 119 DEPs were screened in PC12 cells.Enrichment analysis revealed that DEPs in the rat model were predominantly concentrated in biological functions such as myogenic cell proliferation and signaling related to lipid and energy metabolism.DEPs in the in vitro model were mainly enriched in biological processes such as phagocytosis and were associated with lipid transport and metabolism.Two hub proteins,amyloid precursor protein(APP)and fibronectin 1(FN1),were identified through MCC,MCODE,and Degree scoring.Both PC12 cells and external validation sets showed relatively higher expression of APP and FN1 in injured samples.Results of gene set enrichment analysis indicated that these two proteins were associated with metabolic pathways,such as biosynthesis of glycosaminoglycan chondroitin sulfate and biosynthesis of unsaturated fatty acids.Conclusion:APP and FN1 are potential key molecules involved in SNI and are associated with various metabolic pathways in nerve repair.These findings provide a theoretical basis for the development of therapeutic targets for SNI.

Charcot-Marie-Tooth-1A and sciatic nerve crush rat models:insights from proteomics

The sensorimotor and histological aspects of peripheral neuropathies were already studied by our team in two rat models:the sciatic nerve crush and the Charcot-Marie-Tooth-1A disease.In this study,we sought to highlight and compare the protein signature of these two pathological situations.Indeed,the identification of protein profiles in diseases can play an important role in the development of pharmacological targets.In fact,Charcot-Marie-Tooth-1A rats develop motor impairments that are more severe in the hind limbs.Therefore,for the first time,protein expression in sciatic nerve of Charcot-Marie-Tooth-1A rats was examined.First,distal sciatic nerves were collected from Charcot-Marie-Tooth-1A and uninjured wild-type rats aged 3 months.After protein extraction,sequential window acquisition of all theoretical fragment ion spectra liquid chromatography and mass spectrometry was employed.445 proteins mapped to Swiss-Prot or trEMBL Uniprot databases were identified and quantified.Of these,153 proteins showed statistically significant differences between Charcot-Marie-Tooth-1A and wild-type groups.The majority of these proteins were overexpressed in Charcot-Marie-Tooth-1A.Hierarchical clustering and functional enrichment using Gene Ontology were used to group these proteins based on their biological effects concerning Charcot-Marie-Tooth-1A pathophysiology.Second,proteomic characterization of wild-type rats subjected to sciatic nerve crush was performed sequential window acquisition of all theoretical fragment ion spectra liquid chromatography and mass spectrometry.One month after injury,distal sciatic nerves were collected and analyzed as described above.Out of 459 identified proteins,92 showed significant differences between sciatic nerve crush and the uninjured wild-type rats used in the first study.The results suggest that young adult Charcot-Marie-Tooth-1A rats(3 months old)develop compensatory mechanisms at the level of redox balance,protein folding,myelination,and axonogenesis.These mechanisms seem insufficient to hurdle the progress of the disease.Notably,response to oxidative stress appears to be a significant feature of Charcot-Marie-Tooth-1A,potentially playing a role in the pathological process.In contrast to the first experiment,the majority of the proteins that differed from wild-type were downregulated in the sciatic nerve crush group.Functional enrichment suggested that neurogenesis,response to axon injury,and oxidative stress were important biological processes.Protein analysis revealed an imperfect repair at this time point after injury and identified several distinguishable proteins.In conclusion,we suggest that peripheral neuropathies,whether of a genetic or traumatic cause,share some common pathological pathways.This study may provide directions for better characterization of these models and/or identifying new specific therapeutic targets.

Chitosan conduits enriched with fibrin-collagen hydrogel with or without adipose-derived mesenchymal stem cells for the repair of 15-mm-long sciatic nerve defect

Hollow conduits of natural or synthetic origins have shown acceptable regeneration results in short nerve gap repair;however,results are still not comparable with the current gold standard technique“autografts”.Hollow conduits do not provide a successful regeneration outcome when it comes to critical nerve gap repair.Enriching the lumen of conduits with different extracellular materials and cells could provide a better biomimicry of the natural nerve regenerating environment and is expected to ameliorate the conduit performance.In this study,we evaluated nerve regeneration in vivo using hollow chitosan conduits or conduits enriched with fibrin-collagen hydrogels alone or with the further addition of adipose-derived mesenchymal stem cells in a 15 mm rat sciatic nerve transection model.Unexpected changes in the hydrogel consistency and structural stability in vivo led to a failure of nerve regeneration after 15 weeks.Nevertheless,the molecular assessment in the early regeneration phase(7,14,and 28 days)has shown an upregulation of useful regenerative genes in hydrogel enriched conduits compared with the hollow ones.Hydrogels composed of fibrin-collagen were able to upregulate the expression of soluble NRG1,a growth factor that plays an important role in Schwann cell transdifferentiation.The further enrichment with adipose-derived mesenchymal stem cells has led to the upregulation of other important genes such as ErbB2,VEGF-A,BDNF,c-Jun,and ATF3.

瑞芬太尼调节IL-10/β-内啡肽信号通路对坐骨神经损伤大鼠疼痛的影响

目的探讨瑞芬太尼对坐骨神经损伤(SNI)大鼠疼痛的影响及作用机制。方法建立SNI大鼠模型,大鼠分为假手术组、模型组、瑞芬太尼低剂量组(5μg·kg^(-1)·min^(-1))、瑞芬太尼高剂量组(20μg·kg^(-1)·min^(-1))和瑞芬太尼+抑制剂组(20μg·kg^(-1)·min^(-1)的瑞芬太尼+10μL IL-10抗体),评估大鼠疼痛阈值,统计坐骨神经指数(SFI),Masson染液评价靶肌肉萎缩情况,酶联免疫吸附法(ELISA)检测白介素(IL)-1β、IL-6、肿瘤坏死因子(TNF)-α的含量,免疫荧光检测微管相关蛋白(MAP2)的表达,Western blot检测IL-10、β-内啡肽蛋白表达。结果与假手术组比较,模型组大鼠疼痛阈值、SFI和IL-10、β-内啡肽蛋白的表达水平下降,肌肉萎缩、IL-1β、IL-6、TNF-α、MAP2表达增加(P<0.05);与模型组比较,瑞芬太尼低、高剂量组大鼠疼痛阈值、SFI和IL-10、β-内啡肽蛋白的表达水平升高,肌肉萎缩、IL-1β、IL-6、TNF-α、MAP2表达降低,且呈剂量依赖性(P<0.05);与瑞芬太尼高剂量组比较,瑞芬太尼+抑制剂组大鼠疼痛阈值、SFI和IL-10、β-内啡肽蛋白的表达水平下降,肌肉萎缩、IL-1β、IL-6、TNF-α、MAP2表达增加(P<0.05)。结论瑞芬太尼可能通过激活IL-10/β-内啡肽信号通路抑制SNI大鼠的疼痛行为。

紫铆因通过激活SIRT1介导FOXO1/NF-κB信号通路改善坐骨神经损伤

目的探讨紫铆因诱导SIRT1激活对大鼠坐骨神经损伤的影响及其可能的机制。方法将30只大鼠随机分为假手术组、坐骨神经损伤组和紫铆因组,每组10只。分别于建立大鼠坐骨神经损伤模型手术当天、术后第7天和第14天检测各组大鼠BBB运动评分和坐骨神经功能指数;术后第14天取材,通过HE染色观察各组大鼠坐骨神经病理学改变,通过TUNEL实验检测各组大鼠坐骨神经细胞凋亡水平,通过Western blotting检测各组大鼠坐骨神经BDNF、MBP、GAP-43、SIRT1、FOXO1、Keap1和NF-κB蛋白表达水平。结果与坐骨神经损伤组大鼠相比,紫铆因组坐骨神经损伤大鼠BBB运动评分和坐骨神经功能指数增加,坐骨神经病理损伤改善,坐骨神经细胞凋亡水平降低,坐骨神经BDNF、MBP、GAP-43和SIRT1蛋白表达水平增高,FOXO1、Keap1和NF-κB蛋白表达水平降低。结论紫铆因可通过上调坐骨神经损伤大鼠SIRT1表达抑制FOXO1/NF-κB信号通路激活,继而改善大鼠坐骨神经病理损伤。

齐刺环跳穴干预坐骨神经损伤大鼠脊髓背角炎症因子及胶质原纤维酸性蛋白表达影响的实验研究

目的通过观察齐刺环跳穴对坐骨神经损伤(Sciatic nerve injury,SNI)大鼠脊髓背角炎症因子及胶质原纤维酸性蛋白(Glial fibrillary acidic arotein,GFAP)表达的影响,探讨齐刺环跳穴治疗神经病理痛(Neuropathic pain,NP)的镇痛机制。方法60只SD大鼠随机分为假手术组、模型组、齐刺组、单刺组及药物组,每组12只,采用钳夹法制备大鼠坐骨神经损伤模型。造模第2天开始进行针刺及药物干预,连续14 d。观察各组大鼠干预前后热缩足反射潜伏期(Paw with⁃drawal latency,PWL)的变化,治疗结束后取损伤处坐骨神经,苏木素伊红(HE)染色观察神经形态;取腰3~5脊髓节段ELISA法检测白细胞介素-1β(Interleukin-1β,IL-1β)、白细胞介素-6(Interleukin-6,IL-6)、肿瘤坏死因子-α(Tumor necrosis factor-alpha,TNF-α)蛋白表达水平,实时定量PCR法及免疫组化法检测GFAP表达水平。结果治疗前,与假手术组比较,各组PWL指数降低(P<0.01),与模型组比较,齐刺组治疗后显著改善(P<0.01)。HE染色,模型组神经纤维排列紊乱,齐刺组、单刺组及药物组神经纤维在数量及排列紊乱程度等方面均有改善,齐刺组改善最为明显。ELISA法、RT-PCR法及免疫组化检测,模型组、齐刺组、单刺组、药物组IL-1β、IL-6、TNF-α、GFAP表达较假手术组显著升高(P<0.01);与模型组相比,齐刺组、单刺组、药物组IL-1β、IL-6、TNF-α、GFAP表达显著下降,齐刺组表达低于单刺组及药物组(P<0.01)。结论齐刺环跳穴缓解坐骨神经痛的镇痛机制可能与下调脊髓背角炎症因子表达,抑制星形胶质细胞活化,降低中枢痛觉敏化程度有关。

超声引导下股神经、坐骨神经阻滞在跟骨骨折手术麻醉中的应用效果

目的:探究超声引导下股神经、坐骨神经阻滞在跟骨骨折手术麻醉中的应用效果。方法:选取2020年2月—2023年2月菏泽医学专科学校附属医院收治的跟骨骨折患者80例,以随机数字表法将其均分为对照组(椎管内麻醉)及观察组(超声引导下股神经、坐骨神经阻滞)各40例,对比两组麻醉效果;对比两组麻醉前(T_(0))、麻醉后5 min(T_(1))、麻醉后10 min(T_(2))、麻醉后15 min(T_(3))、麻醉后30min(T_(4))时刻的心率(HR)、平均动脉压(MAP)、血氧饱和度(SpO_(2));对比两组应激反应指标[肾上腺素(E)、皮质醇(Cor)]、凝血功能[凝血酶时间(TT)、凝血酶原时间(PT)、活化部分凝血活酶时间(APTT)]、不良反应发生率。结果:观察组麻醉效果Ⅰ级率高于对照组,差异有统计学意义(P<0.05)。两组T_(0)、T_(1)时刻HR、MAP、SpO_(2)水平比较差异均无统计学意义(P>0.05);对照组T_(2)、T_(3)、T_(4)时刻HR、MAP均高于T_(0)时刻,SpO_(2)均低于T_(0)时刻,差异均有统计学意义(P<0.05);观察组T_(2)、T_(3)、T_(4)时刻HR、MAP、SpO_(2)较T_(0)时刻差异均无统计学意义(P>0.05);观察组T_(2)、T_(3)、T_(4)时刻HR、MAP均低于对照组,SpO_(2)均高于对照组,差异均有统计学意义(P<0.05)。术前,两组E、Cor水平相较差异均无统计学意义(P>0.05);术后1 h两组E、Cor水平均升高,但观察组均低于对照组,差异均有统计学意义(P<0.05)。术前,两组TT、PT、APTT水平相较差异均无统计学意义(P>0.05);术后1 h,两组TT、PT、APTT水平均升高,且观察组均高于对照组,差异均有统计学意义(P<0.05)。观察组不良反应发生率低于对照组,差异有统计学意义(P<0.05)。结论:对跟骨骨折患者实施超声引导下股神经、坐骨神经阻滞,麻醉效果显著,减轻应激反应,稳定术中血流动力学,改善血液高凝状态,降低并发症发生率。

大鼠周围神经损伤后外周血内皮祖细胞动员及相关因子含量变化

目的探讨大鼠周围神经损伤(PNI)后外周血内皮祖细胞(EPCs)、碱性成纤维细胞生长因子(bFGF)、血管内皮生长因子(VEGF)及基质金属蛋白酶-9(MMP-9)水平变化及EPCs与其他指标的相关性。方法42只SD大鼠按随机数字表法分为对照组、PNI 1 d组、PNI 3 d组、PNI 5 d组、PNI 7 d组及PNI 14 d组,每组7只,除对照组外其余组均采用钳夹法建立坐骨神经损伤模型。对每组在预定时间点采用活体心脏穿刺法采血;采用Ficoll密度梯度离心法提取单个核细胞,CD34和CD133双阳性细胞标记EPCs,应用流式细胞仪检测各组EPCs数量。采用酶联免疫吸附试验检测各组外周血bFGF、VEGF及MMP-9含量,分析EPCs数量与bFGF、VEGF、MMP-9水平的相关性。结果与对照组相比,PNI 3 d组、PNI 5 d组、PNI 7 d组外周血EPCs数量升高,PNI 3 d组、PNI 5 d组、PNI 7 d组及PNI 14 d组外周血bFGF含量升高,其余各组外周血VEGF含量升高,PNI 5 d组、PNI 7 d组及PNI 14 d组外周血MMP-9含量升高(P<0.05)。PNI 5 d组和PNI 7 d组外周血EPCs数量与血清bFGF水平呈正相关(r分别为0.784和0.788,P<0.05),与血清VEGF水平呈正相关(r分别为0.889和0.852,P<0.05);PNI 5 d组、PNI 7 d组和PNI 14 d组外周血EPCs数量与血清MMP-9水平呈正相关(r分别为0.788、0.852和0.873,P<0.05)。结论EPCs与bFGF、VEGF和MMP-9共同参与了PNI后血供修复的病理生理过程。

富血小板血浆凝胶缓解CCI模型大鼠周围神经痛及其改善中枢海马组织炎性机制研究

目的建立大鼠坐骨神经慢性压迫损伤(CCI)模型,观察富血小板血浆凝胶(PRP)对CCI大鼠坐骨神经病理性疼痛进展期痛域的影响,并探讨其对中枢海马组织的抗炎作用机制。方法选择SPF级雄性SD大鼠50只,其中10只用于制备PRP,其余40只采用随机数表法分为空白对照组、假手术组、CCI组和CCI+PRP组,每组10只。比较各组大鼠在术前1 d、术后6 h、1 d、3 d、7 d足底机械性缩足反射阈值(MWT)和热辐射缩足潜伏期(TWL)变化;比较术后7 d时各组大鼠海马区肿瘤坏死因子(TNF-α)、白细胞介素-1β(IL-1β)和高迁移率组蛋白1(HMGB1)及其下游Toll样受体-4(TLR4)、糖基化终产物受体(RAGE)mRNA表达水平。结果空白组与假手术组各时间点大鼠MMT和TWL比较差异均无统计学意义(P>0.05);CCI组和CCI+PRP组大鼠术后MMT和TWL值与术前比较差异均有统计学意义(P<0.05);CCI组和CCI+PRP组大鼠术后各时间点MWT和TWL比较差异均有统计学意义(P<0.05)。术后7 d时,对照组和假手术组大鼠海马组织的TNF-α、IL-1β、HMGB1含量以及HMGB1、TLR4和RAGE mRNA表达水平比较差异均无统计学意义(P>0.05),而CCI组和CCI+PRP组与对照组或假手术组比较差异均具有统计学意义(P<0.05);术后7 d时,CCI+PRP组与CCI组大鼠海马组织的TNF-α、IL-1β、HMGB1含量以及HMGB1、TLR4和RAGE mRNA表达水平比较,差异均具有统计学意义(P<0.05)。结论富血小板血浆可有效延缓CCI大鼠神经病理性疼痛进展期痛域,抑制中枢海马组织炎性反应,其机制可能与富血小板血浆通过HMGB1-TLR4/RAGE信号通路抑制TNF-α、IL-1β表达水平有关。

齐刺环跳穴干预坐骨神经损伤大鼠海马IL-1β、IL-6、TNF-α及GFAP表达影响的实验研究

目的 通过观察齐刺环跳穴对坐骨神经损伤大鼠海马白细胞介素-1β(Interleukin-1β,IL-1β)、白细胞介素-6(Interleukin-6,IL-6)、肿瘤坏死因子-α(Tumor Necrosis Factor-alpha, TNF-α)及胶质纤维酸性蛋白(Glial Fibrillary Acidic Protein, GFAP)表达的影响,探讨齐刺环跳穴治疗坐骨神经痛的中枢镇痛机制。方法 60只SD大鼠随机分为假手术组、模型组、齐刺组、单刺组及药物组,每组12只,采用钳夹法制备大鼠坐骨神经损伤模型。造模第2天开始进行针刺及药物干预,连续14 d。观察各组大鼠干预前后坐骨神经功能指数(Sciatic nerve Function Index, SFI)、热缩足反射潜伏期(Paw Withdrawal Latency, PWL)的变化,ELISA法检测海马组织IL-1β、IL-6、TNF-α蛋白表达水平,实时定量PCR法及免疫组化法检测海马组织GFAP表达水平。结果 干预前,与假手术组比较,其余各组大鼠PWL值、SPI值显著降低(P<0.01),与模型组比较,齐刺组干预后大鼠PWL值、SPI值显著改善(P<0.01)。ELISA法、RT-PCR法及免疫组化检测结果显示,模型组、齐刺组、单刺组、药物组IL-1β、IL-6、TNF-α、GFAP表达较假手术组显著升高(P<0.01);与模型组相比,齐刺组、单刺组、药物组IL-1β、IL-6、TNF-α、GFAP表达显著下降,齐刺组表达低于单刺组及药物组(P<0.01)。结论 齐刺环跳穴缓解坐骨神经痛的镇痛机制可能与下调海马炎症因子表达,抑制星形胶质细胞活化,从而降低中枢痛觉敏化程度有关。

坐骨神经胞外核苷酸酶在针刺缓解大鼠踝关节炎性痛中的作用

目的 探讨坐骨神经胞外核苷酸酶[CD39(NTPDase1)]在针刺缓解大鼠踝关节炎性痛中的作用。方法 将54只雄性大鼠随机分为9组,即空白组、踝关节炎性痛模型组、针刺组、坐骨神经腺苷三磷酸(ATP)水解酶抑制(ARL67156注射)+针刺组、坐骨神经CD39上调(三磷酸腺苷双磷酸酶注射)组、坐骨神经CD39特异性抑制(噻氯匹啶注射)+针刺组、坐骨神经CD73特异性抑制(α,β-亚甲基腺苷5′-二磷酸钠盐注射)+针刺组、穴区CD39上调(三磷酸腺苷双磷酸酶注射)组、穴区CD39特异性抑制(噻氯匹啶注射)+针刺组,每组6只。除空白组外,其余各组大鼠均采用注射完全弗氏佐剂建立佐剂性关节炎疼痛模型;造模48 h后,需针刺组大鼠针刺患侧“足三里”穴20 min,需注射药物组于针刺前20 min进行局部相关试剂注射。以大鼠患侧足底机械痛阈值和热痛阈值作为疼痛评价指标;采用实时荧光定量逆转录聚合酶链式反应(RT-qPCR)法和Western blot法检测坐骨神经以及“足三里”穴胞外核苷酸酶的表达。结果 (1)调节坐骨神经ATP水解酶活性:与空白组比较,踝关节炎性痛模型组和坐骨神经ATP水解酶抑制+针刺组热痛阈值和机械痛阈值明显下降(P<0.05);与踝关节炎性痛模型组比较,针刺组热痛阈值和机械痛阈值明显上升(P<0.05)。(2)调节坐骨神经CD39活性:与空白组比较,坐骨神经CD39特异性抑制+针刺组热痛阈值和机械痛阈值明显下降(P<0.05);与踝关节炎性痛模型组比较,针刺组和坐骨神经CD39上调组热痛阈值和机械痛阈值明显上升(P<0.05);与针刺组比较,坐骨神经CD39特异性抑制+针刺组热痛阈值和机械痛阈值明显下降(P<0.05)。(3)调节穴区CD39活性:与空白组比较,穴区CD39特异性抑制+针刺组热痛阈值和机械痛阈值明显下降(P<0.05);与踝关节炎性痛模型组比较,针刺组和穴区CD39上调组热痛阈值和机械痛阈值明显上升(P<0.05);与针刺组比较,穴区CD39特异性抑制+针刺组热痛阈值和机械痛阈值明显下降(P<0.05)。(4)调节坐骨神经CD73活性:与空白组比较,坐骨神经CD73特异性抑制+针刺组热痛阈值和机械痛阈值明显下降(P<0.05);与踝关节炎性痛模型组比较,针刺组热痛阈值和机械痛阈值明显上升(P<0.05)。(5)针刺对坐骨神经以及穴区CD39表达的影响:Western blot法检测结果显示,与空白组比较,踝关节炎性痛模型组坐骨神经CD39的表达显著增加(P<0.05);与踝关节炎性痛模型组比较,针刺组CD39表达显著下调(P<0.05)。RT-qPCR法检测结果显示,与踝关节炎性痛模型组比较,针刺组CD39 mRNA表达水平明显上调(P<0.05)。结论 坐骨神经局部CD39参与了针刺缓解大鼠踝关节炎性痛的机制,这可能与CD73联合促进胞外ATP向腺苷转化有关。

麦粒灸对坐骨神经损伤大鼠脊髓组织TLR4/MyD88/NF-κB信号通路表达的影响

目的观察麦粒灸“环跳”对坐骨神经损伤(SNI)大鼠坐骨神经功能、坐骨神经干病理形态及脊髓组织TLR4/MyD88/NF-κB表达的影响,探究麦粒灸治疗SNI的可能机制。方法24只雄性SD大鼠随机分为空白组、假手术组、模型组和麦粒灸组,每组6只。模型组、麦粒灸组采用坐骨神经钳夹制备SNI大鼠模型,造模后第7日起麦粒灸组取患侧“环跳”麦粒灸干预,每次6壮,1次/d,连续10 d。观察造模后第7日及干预结束后大鼠坐骨神经功能指数(SFI)、纤维丝测痛仪测量大鼠机械痛阈值(MWT),ELISA检测脊髓组织一氧化氮(NO)、诱导型一氧化氮合酶(iNOS)含量,HE染色观察坐骨神经干形态,Western blot检测脊髓组织Toll样受体4(TLR4)、核因子(NF)-κBp65、p-NF-κBp65、髓样分化因子88(MyD88)、NF-κB抑制蛋白α(IκBα)、p-IκBα蛋白表达。结果与假手术组比较,模型组大鼠SFI、MWT显著降低(P<0.01),坐骨神经干神经纤维排列紊乱,施万细胞数量明显增多,有大量空泡变性,脊髓组织NO、iNOS含量及TLR4、p-NF-κBp65、MyD88、p-IκBα蛋白表达显著升高(P<0.01);与模型组比较,麦粒灸组大鼠SFI、MWT显著升高(P<0.01),坐骨神经干损伤减轻,细胞排列较整齐,施万细胞数量减少,轴突脱髓鞘及细胞空泡变性减少,脊髓组织NO、iNOS含量及TLR4、p-NF-κBp65、MyD88、p-IκBα蛋白表达显著降低(P<0.05)。结论麦粒灸“环跳”可下调SNI大鼠脊髓组织TLR4、p-NF-κBp65、MyD88、p-IκBα蛋白表达,抑制NO、iNOS分泌,从而缓解疼痛、减轻受损神经组织炎症反应。

法舒地尔促进大鼠坐骨神经横断伤后的轴突与髓鞘再生及功能恢复

目的 探讨法舒地尔在改善周围神经损伤后的轴突与髓鞘再生及功能恢复的效果。方法SD大鼠30只,重(200±30)g,切断右侧坐骨神经,缝合后等分入2组,即对照组与法舒地尔组,分别给予生理盐水与10mg/kg的盐酸法舒地尔腹腔注射。术后2周,利用NF-200一抗对损伤远端的轴突密度进行评估。术后4周,利用逆行示踪剂荧光金评估发出轴突进入损伤远端的位于L4~6DRG和腰骶膨大处的神经元数量。术后12周,对腓肠肌的湿重比、肌纤维的横截面积进行测量;利用NF-200与MPZ一抗及透射电镜对轴突的直径与髓鞘的厚度进行测量。术后4、6、8、10、12周采集足印,对两组大鼠坐骨神经功能指数(SFI)进行评定。此外,体外培养大鼠胚胎脊髓背根神经节(DRG)和脊髓运动神经元(SMN),评估法舒地尔对其轴突生长的影响。结果 术后14 d,法舒地尔组损伤远端神经密度显著高于对照组(P=0.034)。术后4周法舒地尔组被荧光金逆行标志的L4~6DRG及脊髓前角运动神经元数目均显著高于对照组(P<0.05)。术后12周,法舒地尔组有髓轴突的数量、有髓轴突的直径以及髓鞘的厚度都高于对照组(P<0.05),G-ratio值则低于对照组(P<0.05)。SFI数据结果显示,术后6、8、10及12周,法舒地尔组SFI值显著优于对照组(P<0.05)。术后12周,对照组的患侧腓肠肌的湿重比及肌纤维的横截面积显著小于法舒地尔组(P<0.01)。培养5 d后,法舒地尔组DRG及SMN的轴突显著长于对照组(P<0.01)。结论 法舒地尔能够促进大鼠坐骨神经横断伤后的轴突与髓鞘再生及其运动功能恢复。

基于SP/NK1R/βARRS通路电针缓解神经性疼痛的实验研究

目的观察针刺对坐骨神经慢性缩窄模型(chronic constriction injury,CCI)大鼠疼痛行为及脊髓背角P物质(substance P,SP)含量和神经激肽1受体(neurokinin 1 receptor,NK1R)、β-抑制蛋白1抗体(β-arrestin 1,βARR1)蛋白水平的影响。方法将36只雄性Sprague-Dawley(SD)大鼠随机分为假手术组、模型组、电针组和手针组,每组9只。暴露模型组、电针组和手针组大鼠左侧股骨中段坐骨神经并进行结扎,建立坐骨神经慢性缩窄模型;假手术组暴露坐骨神经但不结扎。造模后第8天开始,电针组和手针组分别对环跳和阳陵泉穴进行干预。测定4组大鼠造模后第0、7、13、21、29天的机械痛阈、热痛阈及后肢负重分布。采用酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)检测4组大鼠L4~L6腰膨大脊髓背角组织中SP含量;用蛋白质印迹法(Western blotting)检测4组大鼠L4~L6腰膨大脊髓背角组织中NK1R和βARR1蛋白表达。结果造模后第7天,模型组、电针组和手针组大鼠机械痛阈和热痛阈较假手术组降低(P<0.05),两后肢负重分布差异较假手术组增加(P<0.05)。针刺干预后,造模后第13、21、29天,手针组和电针组大鼠机械痛阈和热痛阈高于模型组(P<0.05),两后肢负重分布差异小于模型组(P<0.05);造模后第29天,电针组机械痛阈和热痛阈均高于手针组(P<0.05);造模后第21天,电针组两后肢负重分布差异小于手针组(P<0.05)。针刺干预全部结束后,模型组脊髓背角组织SP含量低于假手术组(P<0.05),手针组和电针组SP含量高于模型组(P<0.05)。针刺干预全部结束后,模型组脊髓背角组织NK1R和βARR1蛋白表达高于假手术组(P<0.05),电针组和手针组NK1R和βARR1蛋白表达低于模型组(P<0.05)。结论针刺可能通过调控脊髓背角SP水平及NK1R和βARR1蛋白表达对神经病理性疼痛起到镇痛效应。