神经病理性痛大鼠脊髓背角神经元凋亡及Bax和Bcl-2的表达

早期的组织学研究表明外周神经损伤后脊髓背角抑制性中间神经元可发生跨突触变性死亡，近年的研究采用末端生脱氧核糖核苷酸转移酶介导生物素标记（TUNEL）技术证实凋亡是外周神经损伤后脊髓神经元死亡的重要方式。Bax和Bcl-2是一对作用相反的凋亡调控蛋白，在病理状态下神经元凋亡中发挥重要的调控作用。本研究拟观察神经病理性痛大鼠脊髓背角神经元凋亡及凋亡相关蛋白Bax和Bcl-2的表达。     

氯胺酮对慢性坐骨神经压迫性损伤大鼠脊髓背角神经元凋亡及Bax和Bcl-2的表达影响

目的观察氯胺酮（Ket）对慢性坐骨神经压迫性损伤（CCI）大鼠脊髓背角神经元凋亡及凋亡相关蛋白Bax和Bcl-2表达的影响。方法选择雄性SD大鼠40只，随机分为4组，分别为空白对照组，假手术组，CCI组和腹腔注射Ket 50mg／kg治疗组。假手术组，CCI组和Ket治疗组又按术后取材时间的不同分别分为3个亚组，即术后3d组、术后9d组和术后14d组。用TUNEL标记法标记脊髓背角浅层凋亡细胞。应用免疫组织化学方法和免疫印迹（Western-blot）法检测大鼠脊髓背角浅层神经元凋亡相关蛋白Bax和Bcl-2表达情况。结果与空白对照组和假手术组各亚组比较，CCI3d，9d，14d亚组和Ket9d，14d亚组大鼠脊髓背角的凋亡细胞明显增多（P〈0．01）；与CCI3d亚组比较，Ket3d亚组大鼠脊髓背角的凋亡细胞明显减少（P〈0．01）。与空白对照组和假手术各亚组比较，CCI3d，9d，14d亚组大鼠脊髓背角浅层Bax阳性神经元和Bax的表达显著增加（P〈0．01），CCI9d，14d亚组大鼠脊髓背角浅层Bcl-2阳性神经元和Bcl-2的表达明显增加（P〈0．01）。与CCI3d亚组比较，Ket3d亚组脊髓背角浅层Bax阳性神经元和Bax的表达显著减少（P〈0．01），而Bcl-2阳性神经元和Bax的表达显著增加（P〈0．01）。结论Ket能抑制脊髓背角浅层神经元Bax的表达，促进Bcl-2的表达，提示Ket对神经病理性疼痛状态下脊髓背角神经元的凋亡具有保护作用。     

蛛网膜下腔注射氯胺酮对慢性神经病理痛大鼠的镇痛机制

外周神经损伤后,会出现痛觉异常,如自发痛、痛觉过 敏、异源痛等,可能与脊髓背角浅层神经元凋亡有关,有研 究表明蛛网膜下腔注射氯胺酮可以减轻神经病理痛。但 其作用机制尚不清楚。本研究拟通过观察蛛网膜下腔注射 氯胺酮对慢性神经病理痛大鼠脊髓背角浅层细胞凋亡的影 响,探讨其对神经病理痛的镇痛机制。     

γ-氨基丁酸及其受体与脊髓伤害性信息传递

脊髓特别是脊髓背角是伤害性信息从感觉传入纤维传入中枢神经系统(CNS)进行整合和传递的第一站，其中存在着相互拮抗的两种系统，即兴奋性和抑制性调节系统，二者对多种因素(组织和神经损伤、炎症、缺血)导致的疼痛传递起着重要的作用。GABA递质／受体系统是脊髓内重要的抑制性调节系统，参与伤害性信息传递的机制，受到人们关注。     

生长抑素中间神经元与认知功能关系的研究进展

神经网络是学习与记忆等认知功能的生物学基础,而兴奋与抑制功能失衡很可能是神经精神疾病发生的重要病理机制。其中,γ-氨基丁酸(GABA)能抑制性中间神经元及其抑制性神经环路在神经网络形成中起重要调节作用。越来越多的研究表明,生长抑素(somatostatin,SOM)中间神经元参与调控神经网络活动并在认知功能中发挥关键作用。本文主要就SOM中间神经元在认知功能及相关疾病中的作用作一综述,以期为认知功能相关的研究提供参考。     

NGF和BDNF在炎性痛大鼠背根神经节和脊髓背角的表达

目的 观察神经生长因子(NGF)和脑源性神经营养因子(BDNF)在福尔马林致痛大鼠外周、背根神经节(DRG)和脊髓背角的表达变化及相互关系，探讨神经营养因子在疼痛发生机制中的作用。方法 采用大鼠后足掌皮下注射福尔马林溶液的外周炎性痛模型。成年SD大鼠24只随机分为四组(每组6只)：对照组(A组)；致痛组根据取材时间不同分为，致痛后2h取材(B组)、致痛后24h取材(C组)、致痛后48h取材(D组)。观察致痛1h内疼痛行为变化，应用免疫组织化学方法结合计算机图像分析技术检测致炎足底皮肤、DRG和脊髓背角浅层内NGF的表达，及DRG和脊髓背角浅层表达BDNF的变化。结果 所有致痛组大鼠都表现出明显的两期伤害性反应。在致痛后2h患侧足底皮肤、DRG和24h时脊髓背角浅层表达的NGF开始增加，致痛24h后BDNF在患侧DRG表达开始增加，且与NGF在患侧DRG神经元的表达变化成正相关。48h时BDNF在患侧脊髓背角浅层表达开始增加。结论 NGF不仅是产生炎性痛外周机制的重要介质之一，还可通过促进BDNF在脊髓背角的表达，共同参与中枢敏感化的产生。     

糖皮质激素受体在慢性吗啡耐受大鼠脊髓背角神经元凋亡中的作用

目的 评价糖皮质激素受体在慢性吗啡耐受大鼠脊髓背角神经元凋亡中的作用.方法 鞘内置管成功的健康雄性SD大鼠20只,体重300～350 g,随机分为4组(n=5):对照组(C组)、慢性吗啡耐受组(M组)、吗啡+糖皮质激素受体拮抗剂组(MR组)和吗啡+糖皮质激素受体激动剂组(MD组)分别于8:00和20:00鞘内注射生理盐水10μl、吗啡10μg、吗啡10μg+RU38486 2μg、吗啡10μg+地塞米松4μg,连续6 d.于每天8:00给药后30 min行甩尾实验,给药第7天处死大鼠,取L3～L5脊髓行TUNEL染色,光镜下观察脊髓背角神经元的凋亡情况,计算凋亡率.结果 地塞米松、RU38486分别对慢性吗啡耐受的形成起促进、抑制作用.与C组比较,M组和MD组脊髓背角神经元凋亡率升高(P＜0.05);与M组比较,MR组脊髓背角神经元凋亡率降低,MD组脊髓背角神经元凋亡率升高(P＜0.05).结论 糖皮质激素受体参与了慢性吗啡耐受形成中大鼠脊髓背角神经元凋亡的过程.     

右美托咪啶对慢性神经病理性痛大鼠脊髓背角神经元凋亡的影响

目的 评价右美托咪啶对神经病理性痛大鼠脊髓背角神经元凋亡的影响.方法 健康成年雄性SD大鼠72只,体重180～220 g,采用随机数字表法,将其随机分为3组(n=24):假手术组(S组)、慢性神经病理性痛组(CNP组)和右美托咪啶组(D组).S组仅分离坐骨神经但不结扎,CNP组和D组采用结扎坐骨神经的方法制备大鼠神经病理性痛模型,D组于结扎坐骨神经结束开始至处死前,腹腔注射右美托咪啶50μg/kg,1次/d,S组和CNP组注射等容量生理盐水.于术前1d、术后3、7、14 d(T0-3)时测定大鼠机械缩足阈值(MWT)和热缩爪潜伏期(TWL);于T1-3时测定痛阈后每组随机处死8只大鼠,取L4,5脊髓组织,采用免疫组化法检测脊髓背角Bcl-2及caspase-3的表达水平,采用透射电镜观察脊髓背角浅层神经元超微结构.结果 与S组比较,CNP组和D组T1-3时MWT降低,TWL缩短,脊髓背角Bcl-2和caspase-3表达上调(P＜0.05);与CNP组比较,D组T1 -3时MWT升高,TWL延长,脊髓背角Bcl-2表达上调,caspase-3表达下调(P＜0.05).脊髓背角浅层神经元超微结构:S组基本正常,CNP组细胞凋亡数目增加,D组细胞凋亡数目较CNP组减少.结论 腹腔注射右美托咪啶可减轻大鼠慢性神经病理性痛,其机制可能与抑制脊髓背角神经元凋亡有关.     

神经肽Y2受体在大鼠神经病理性痛中的作用

目的 探讨神经肽Y2受体(NPY2R)在大鼠神经病理性痛中的作用.方法 SPF级雄性SD大鼠36只,8周龄,体重190～ 210 g,采用随机数字表法,将其分为3组(n=12):假手术组(S组)、神经病理性痛组(NP组)和NPY2R反义寡核苷酸组(ODN组).NP组和ODN组采用坐骨神经慢性压迫法制备神经病理性痛模型.术后7d时ODN组鞘内注射5μg/μlNPY2R反义寡聚核苷酸30 m.分别于术前3 d(T0,基础状态)、术后7 d(T1)、鞘内给药后15 min、1.5、3.0、4.5、6.0 h(T2-6)时测定机械痛阈和冷痛阈,然后处死大鼠,取L4-6脊髓组织,采用免疫荧光法测定脊髓背角神经元NPY2R、降钙素基因相关肽(CGRP)的表达和二者共表达(NPY2R/CGRP)水平.结果 与S组比较,NP组和ODN组T1.时机械痛阈降低,冷痛阈升高,脊髓背角神经元NPY2R、CGRP表达上调(P＜0.05);与NP组比较,ODN组T3-5时机械痛阈升高,脊髓背角神经元NPY2R和NPY2R/CGRP表达下调(P＜0.05),冷痛敏和脊髓背角神经元CGRP表达差异无统计学意义(P＞0.05).结论 脊髓背角神经元NPY2R参与了大鼠神经病理性痛的机械痛觉过敏维持,而未参与冷痛觉过敏维持.     

去甲肾上腺素和麻醉镇痛

在具有完整的中枢去甲肾上腺素(NA)能神经系统的前提下,NA通过作用于中枢α1或(和)α2受体来影响不同麻醉药的镇痛作用,破坏中枢NA能神经系统的完整性亦如此。脊髓NA能神经系统在麻醉药的镇痛机制中发挥着重要作用。在脊髓水平,NA可通过α1受体使脊髓胶质层抑制性神经元去极化而促进γ-氨基丁酸及甘氨酸的释放;经α2受体抑制脊髓初级传入纤维末梢释放谷氨酸和P物质,并使脊髓背根神经节细胞和脊髓胶质层神经元超极化来增强或介导麻醉药的镇痛作用。     

Opioidinduzierter Pruritus

Substantial progress has been achieved in recent years in research on the interaction between pain and pruritus. Over and above the known inhibition of pruritus by painful stimuli (e.g. scratching), a foundation for the explanation of opioid-induced pruritus was laid through the discovery of pruritus-specific neuronal processing channels. Although traditionally the degranulating effect of opioids on mast cells was assumed to be the essential mechanism, it is now clear that opioids can also induce itching at the spinal level. Neurons of the dorsal horn of the pain system inhibit spinal itch neurons. If this inhibition is weakened by opioids, the disinhibited itch neurons become active and mediate itching, without stimulation of the primary afferent peripheral nerves. Spinal triggering of itching is observed in particular by activation of µ-opioid receptors (μ-OR), while κ-OR surprisingly suppress itch. The therapeutic implications of this interaction will be described.     

Autonomic dysreflexia, induced by noxious or innocuous stimulation, does not depend on changes in dorsal horn substance p

After experimental spinal cord injury (SCI) in rats, autonomic dysreflexia is commonly induced by slightly noxious cutaneous or visceral stimuli. The presence of autonomic dysreflexia is associated with an increase in the afferent fiber arbor area labeled by cholera toxin B or with an anti-CGRP antibody. Our goal was to examine further the sensory afferent input contributing to exaggerated autonomic spinal reflexes and subsequent increases in blood pressure after SCI, typical of autonomic dysreflexia. We observed that changes in blood pressure and heart rate induced by slightly noxious stimuli (2.0-mL balloon colon distension, cutaneous pinch) were increased in magnitude with time after SCI. In contrast, cardiovascular responses induced by non-noxious stimuli (1.0-mL balloon colon distension, light stroking of hair) were relatively constant. We examined substance P-immunoreactive afferent fibers to identify type C, unmyelinated afferent fibers, and A delta lightly myelinated fibers in superficial and deeper laminae of the dorsal horn, respectively. The area of substance P-immunoreactive fibers was quantified in laminae I-V of the dorsal horn. Analysis revealed no difference in substance P afferent fiber area in laminae I-II, or laminae III-V, between sham-injured and SCI rats. These data suggest that noxious, or innocuous, stimulation induces autonomic dysreflexia without expansion of the central arbors of substance P-immunoreactive sensory neurons. Furthermore, autonomic dysreflexia induced by noxious stimulation increases with time after spinal cord injury.     

Effect of dorsal-column stimulation on gelatinosa and marginal neurons of cat spinal cord

Single neuronal units with physiological characteristics of superficial dorsal-horn neurons were recorded extracellularly in laminae 1, 2, and 3 of cat spinal cord. When focal electrical stimulation was applied to the ipsilateral dorsal column, most of the units were excited transsynaptically at various latencies consistent with an effect mediated by large myelinated axons. Units recorded in laminae 2 and 3 had earlier latencies of activation than units in lamina 1. Units with cutaneous receptive fields only for noxious stimuli were activated at significantly longer latencies than units responsive to innocuous stimuli. The time course of these effects was consistent with the concept that many cells in laminae 1 to 3 receive direct excitatory synaptic input from collaterals of dorsal-column fibers, and some lamina 1 cells receive excitatory synaptic input from lamina 2 neurons. Previous reports have emphasized the inhibitory action of dorsal-column stimulation on nociceptive responses of cells in laminae 4 and 5 of the dorsal-horn, particularly those of the spinocervical tract in cats and the spinothalamic tract in primates. The present study suggests that some of this inhibition might be sustained by a network of interneurons in or near the substantia gelatinosa and marginal layer. The therapeutic efficiency of dorsal-column stimulation for pain relief in humans may depend in part on the activation of neurons in the superficial layers of the dorsal horn.     

Meperidine suppresses the excitability of spinal dorsal horn neurons

BACKGROUND: In addition to local anesthetics, meperidine has been successfully used for local anesthesia. When applied intrathecally, the dorsal horn neurons of the superficial laminae are exposed to high concentrations of meperidine. These cells represent an important point for the transmission of pain information. This study investigated the blocking effects of meperidine on different ionic currents of spinal dorsal horn neurons and, in particular, its impact on the generation of action potentials. METHODS: Using a combination of the patch clamp technique and the entire soma isolation method, the action of meperidine on voltage-gated Na+ and K+ currents in spinal dorsal horn neurons of rats was described. Current clamp recordings from intact neurons showed the functional relevance of the ion current blockade for the generation of action potentials. RESULTS: Externally applied meperidine reversibly blocked voltage-gated Na+ currents with a half-maximum inhibiting concentration (IC50) of 112 microM. During repetitive stimulation, a slight phasic block occurred. In addition, A-type K+ currents and delayed-rectifier K+ currents were affected in a dose-dependent manner, with IC50 values of 102 and 52 microM, respectively. In the current clamp mode, single action potentials were suppressed by meperidine. The firing frequency was lowered to 54% at concentrations (100 microM) insufficient for the suppression of a single action potential. CONCLUSIONS: Meperidine inhibits the complex mechanism of generating action potentials in spinal dorsal horn neurons by the blockade of voltage-gated Na+ and K+ channels. This can contribute to the local anesthetic effect of meperidine during spinal anesthesia.     

Clonidine-induced neuronal activation in the spinal cord is altered after peripheral nerve injury

BACKGROUND: Alpha 2 adrenoceptor agonists produce antinociception in normal animals and alleviate mechanical allodynia in animals with nerve injury, although their mechanism of action may differ in these situations. The purpose of this study was to examine the location and number of cells in the spinal cord activated by intrathecal clonidine in these two circumstances and to test whether one class of interneurons, cholinergic, express alpha 2 adrenoceptors. METHODS: Intrathecal saline or clonidine, 10 and 30 microg, was injected in normal rats or those with mechanical allodynia following partial sciatic nerve section. Two hours later, animals were anesthetized and pericardially perfused. The number of cells in superficial and deep dorsal horn laminae at the L4-L5 level immunostained for phosphorylated cAMP response element binding protein (pCREB) were quantified. In separate studies, the authors colocalized alpha2C adrenoceptors with cholinergic neurons. RESULTS: Intrathecal clonidine increased pCREB immunoreactive cells in both superficial and deep laminae by 50-100% in normal animals. The number of pCREB immunoreactive cells increased in nerve-injured compared to normal rats. Intrathecal clonidine decreased pCREB immunoreactive cells in the deep dorsal horn of injured animals. Alpha2C adrenoceptors colocalized with cholinergic neurons in both superficial and deep dorsal horn. DISCUSSION: Previous studies suggest a shift in alpha 2 adrenoceptor subtype and the involvement of cholinergic interneurons in antinociception in the spinal cord after nerve injury. The current results suggest that intrathecal clonidine, by direct or indirect methods, increases neuronal activation in normal animals, presumably leading to net inhibition of pain signaling, whereas it reduces the increase in neuronal activity induced by nerve injury.     

Neuraxial opioid-induced pruritus: a review

When intrathecal and epidural opioids are administered, pruritus occurs as an unwanted and troublesome side effect. The reported incidence varies between 30% and 100%. The exact mechanisms of neuraxial opioid-induced pruritus remain unclear. Postulated mechanisms include the presence of an "itch center" in the central nervous system, medullary dorsal horn activation, and antagonism of inhibitory transmitters. The treatment of intrathecal opioid-induced pruritus remains a challenge. Many pharmacological therapies, including antihistamines, 5-HT(3)-receptor antagonists, opiate-antagonists, propofol, nonsteroid antiinflammatory drugs, and droperidol, have been studied. In this review, we will summarize pathophysiological and pharmacological advances that will improve understanding and ultimately the management of this troublesome problem.     

Exposure of the dorsal root ganglion in rats to pulsed radiofrequency currents activates dorsal horn lamina I and II neurons

OBJECTIVE: Application of pulsed radiofrequency (RF) currents to the dorsal ganglion has been reported to produce long-term relief of spinal pain without causing thermal ablation. The present study was undertaken to identify spinal cord neurons activated by exposure of the dorsal ganglion to pulsed RF currents in rats. METHODS: Left-sided hemilaminectomy was performed in adult Sprague-Dawley rats to expose the C6 dorsal root ganglion. An RF electrode (0.5 mm diameter) with a thermocouple for temperature monitoring was positioned on the exposed ganglion, and rats were assigned to one of three treatment groups: pulsed RF treatment (20 ms of 500-kHz RF pulses delivered at a rate of 2 Hz for 120 s to produce tissue heated to 38 degrees C), continuous RF (continuous RF currents for 120 s to produce tissue heated to 38 degrees C), or sham treatment (no RF current; electrode maintained in contact with ganglion for 120 s). RESULTS: Treatment with pulsed RF but not continuous RF was associated with a significant increase in the number of cFOS-immunoreactive neurons in the superficial laminae of the dorsal horn as observed 3 hours after treatment. CONCLUSION: Exposure of the dorsal ganglion to pulsed RF currents activates pain-processing neurons in the dorsal horn. This effect is not mediated by tissue heating.     

Effect of electrical stimulation of peripheral nerves on neuropathic pain

STUDY DESIGN: Changes in the electrophysiologic response of spinal dorsal horn neurons elicited by peripheral electrical stimulation were examined. OBJECTIVE: To investigate whether the electrical stimulation of peripheral nerves causes an inhibition of pain at the spinal cord level. SUMMARY OF BACKGROUND DATA: The wide dynamic range neurons studied were known to be excited by primary afferent fibers, not only combined A (delta) and C nociceptive fibers, but also low-threshold mechanoreceptive A (beta) fibers and A (delta) fibers of down hairs. The wide dynamic range neurons are classified as nociceptive neurons. METHODS: Responses of wide dynamic range neurons in the lumbosacral dorsal horn to input from C fibers were studied in urethane chloralose-anesthetized cats. The posterior tibial nerve and sciatic nerve were stimulated simultaneously to examine the effect on the C fiber responses elicited by superficial peroneal nerve stimulation. RESULTS: Simultaneous stimulation of the posterior tibial nerve and sciatic nerve was performed with superficial peroneal nerve C fiber stimulation. CONCLUSIONS: This study demonstrated that electrical stimulation of peripheral nerves leads to inhibitory input to the pain pathways at the spinal cord level.     

Lichen simplex chronicus as the initial manifestation of intramedullary neoplasm and syringomyelia.

Neurogenic causes of pruritus and a rash are uncommon. We report a patient with dermatomal pruritus and a rash who had a cervicothoracic syrinx and a thoracic spinal cord tumor. We believe the syrinx interrupted fibers subserving itch, resulting in dermatomal pruritus with secondary scratching and a rash.     

Anatomy,physiology and pharmacology of pain

Nociception is conveyed from the periphery to the brain at three levels: the peripheral nociceptor, the spinal cord, and the supra-spinal (brain) levels. Physiological (first or ‘fast’) pain is produced by stimulation of high threshold thermo/mechanical nociceptors, which transmit via fast conducting myelinated A delta fibres. These enter the dorsal horn of the spinal cord and synapse at laminae I and V. Pathophysiological (second or ‘slow’) pain originates from stimulation of the high threshold polymodal nociceptors (free endings) present in all tissues. The nociceptors respond to mechanical, chemical and thermal stimuli and are transmitted via slow conducting unmyelinated C fibres. These synapse at laminae II and III (substantia gelatinosa) of the dorsal horn. The second order neurons are either nociceptive specific (substantia gelatinosa) or wide dynamic range (WDR) neurons (in laminae V and VI) that respond to a wide range of noxious and non-noxious input. Both pathways ascend up the spinal cord via the spinothalamic tracts to the thalamus, which synapse and project on to the somatosensory cortex. Inhibitory inter-neurons in the substantia gelatinosa prevent activation of the dorsal root ganglia. Interneurons can be activated by A beta and inhibited by A beta and C fibre activity. Pain can be ‘gated-out’ by stimulating the large A beta fibres in the painful area. This is the working mechanism behind transcutaneous electrical nerve stimulation. The descending inhibition pathways originate at the level of the cortex and thalamus, and descend via the brainstem (periaqueductal grey) and the dorsal columns to terminate at the dorsal horn of the spinal cord. Neurotransmitters noradrenaline, serotonin (5-HT) and the endogenous opioids are released to provide antinociception.