胶质细胞活化在病理性疼痛发展过程中的意义：对药物研发的启示

近年来有关胶质细胞在病理性疼痛发生和发展过程中的作用越来越受到了重视。目前认为小胶质细胞和星形胶质细胞在外周神经或组织损伤后被激活并产生各种炎性介质,成为病理性疼痛发生发展过程中重要的致痛因素,针对胶质细胞活化及其后产生的炎性介质有可能研发出新型抗病理性疼痛药物。本文就此方面的研究进展做一综述,试图为靶向胶质细胞研发新型抗病理性疼痛药物提供新的线索。     

京尼平苷对糖尿病大鼠神经病理性疼痛的影响

目的探讨京尼平苷对链脲佐菌素诱导的1型糖尿病性神经病理性疼痛大鼠的镇痛作用及可能作用机制。方法通过链脲佐菌素诱导的SD大鼠建立糖尿病性神经病理性疼痛模型,将大鼠随机分为对照组、模型组、阳性对照组(加巴喷丁)及不同剂量京尼平苷处理组(1,10和100mg·kg-1)。建模后第21天给予不同剂量京尼平苷,连续给药7d。免疫荧光组织化学法观察脊髓背角小胶质细胞活化情况,ELISA法观察脊髓致炎细胞因子水平。结果连续给予京尼平苷对糖尿病大鼠机械痛敏具有较好的镇痛作用,同时观察到京尼平苷可显著抑制脊髓背角小胶质细胞活化及致炎细胞因子TNF-α和IL-1β水平;同时京尼平苷可降低糖尿病大鼠的血糖水平。结论京尼平苷对糖尿病性神经病理性疼痛具有较好的镇痛作用,其机制可能与抑制脊髓背角小胶质细胞活化及致炎细胞因子水平有关。     

星形胶质细胞的免疫炎性反应和谷氨酸代谢异常在卒中后抑郁发病中的作用

星形胶质细胞是人脑内数量最多的细胞,在缺血早期结构会发生明显变化,表达多种神经营养因子和损伤因子。卒中发生后,星形胶质细胞活化为反应性星形胶质细胞,通过神经保护或神经毒性作用影响组织修复和神经炎性疾病的发生和发展。卒中诱发细胞间谷氨酸浓度及炎性因子的增加会促进卒中后抑郁发生,而卒中后活化的星形胶质细胞会通过多条通路调控免疫炎性反应和神经元谷氨酸代谢。现就星形胶质细胞介导的免疫炎性反应与谷氨酸能系统对卒中后抑郁中发生发展信号通路做进一步剖析,以期提供靶向调控星形胶质细胞调节谷氨酸及炎性因子水平策略,为治疗卒中后抑郁提供思路和线索。     

京尼平苷对LPS诱导的星形胶质细胞活化及炎性反应的抑制作用

目的明确京尼平苷对脂多糖(LPS)诱导的星形胶质细胞活化及炎性反应的抑制作用。方法以LPS诱导原代培养的星形胶质细胞为细胞模型,随机分为对照组、LPS组、LPS+白藜芦醇组和LPS+不同浓度京尼平苷组,采用Western Blot法检测星形胶质细胞活化标记物GFAP水平及NF-κB p65水平,ELISA法检测致炎细胞因子水平。结果京尼平苷可显著抑制LPS诱导的星形胶质细胞活化,降低TNF-α、IL-1β和IL-6的表达,同时NF-κB p65蛋白的表达也明显降低。结论京尼平苷能通过NF-κB途径抑制LPS诱导的星形胶质细胞活化,并抑制致炎细胞因子水平。     

鞘内注射氟代柠檬酸对糖尿病大鼠神经病理性疼痛的影响

研究发现脊髓的星形胶质细胞在疼痛的发生和维持中起到十分重要作用[1～3].氟代柠檬酸(flourocltrate,FC)是星形胶质细胞的特异性抑制剂,它在脊神经损伤模型中能抑制星形胶质细胞过渡活化,减轻神经病理性疼痛[2,3].但氟代柠檬酸对糖尿病大鼠神经病理性疼痛的影响如何尚未见研究.本研究观察氟代柠檬酸对糖尿病大鼠神经病理性疼痛模型大鼠痛行为及脊髓星形胶质细胞的影响,以探讨其机制.     

阿魏酸对小胶质细胞炎性反应的抑制作用

目的考察阿魏酸对脂多糖诱导的小鼠小胶质细胞炎性反应的抑制作用及其机制。方法采用脂多糖(LPS)刺激小胶质细胞(BV-2)活化,研究阿魏酸对炎症反应的抑制作用。采用硝酸还原酶法检测阿魏酸对一氧化氮(nitric oxide,NO)的影响,定量PCR和蛋白印迹分析阿魏酸对诱导型一氧化氮合酶(inducible nitric oxide synthase,i NOS)和环氧合酶-2(COX-2)的影响,定量PCR技术和ELISA分析阿魏酸对炎性因子白细胞介素-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-α,TNF-α)的影响,进而检测阿魏酸对促分裂原活化蛋白激酶(mitogen-activated protein kinases,MAPK)信号通路的影响。结果 1.25~20μmol·L-1的阿魏酸对细胞活性无明显影响。阿魏酸浓度依赖性降低NO的浓度,可以明显抑制i NOS、COX-2的基因和蛋白表达,明显抑制IL-1β、IL-6、TNF-α的表达,提前给予阿魏酸共同孵育对LPS引起的ERK信号通路的磷酸化有抑制作用。结论阿魏酸具有抑制小胶质细胞活化,抑制神经性炎症的作用,其机制可能是阿魏酸通过ERK信号通路发挥对炎性分子的抑制作用。     

复方环磷腺苷乳膏对银屑病小鼠模型的作用

目的:研究复方环磷腺苷乳膏对银屑病小鼠模型的治疗作用。方法:采用小鼠阴道上皮和小鼠尾鳞片上皮银屑病样皮损模型,观察复方环磷腺苷乳膏对上皮细胞分裂及表皮细胞分化的影响。结果:复方环磷腺苷乳膏能明显抑制阴道上皮细胞有丝分裂,对小鼠尾鳞片表皮颗粒层形成有显著促进作用。结论:复方环磷腺苷乳膏为治疗银屑病的有效药物。     

复方环磷腺苷乳膏治疗银屑病的药效学研究

目的研究复方环磷腺苷乳膏对银屑病动物模型的治疗作用。方法采用小鼠阴道上皮病理模型、小鼠尾鳞片上皮病理模型和豚鼠耳部皮肤银屑病样皮损模型,观察复方环磷腺苷乳膏对上皮细胞分裂及表皮细胞分化的影响。结果复方环磷腺苷乳膏能明显抑制阴道上皮细胞有丝分裂、对小鼠尾鳞片表皮颗粒层形成有显著促进作用、使豚鼠银屑病样模型耳部皮肤厚度显著减少。结论复方环磷腺苷乳膏为治疗银屑病的有效药物。     

复方中药塞络通对多发性脑梗死大鼠星形胶质细胞功能及脂笼蛋白2蛋白表达的影响

目的脑卒中是一种常见的疾病,绝大多数脑卒中病例是由短暂性或永久性脑血管闭塞引起的缺血性脑血管病(缺血性中风)最终导致脑梗死。星形胶质细胞做为脑内大量存在的细胞,在缺血损伤后形态肥大,表现为突起延长和胞体肿胀。脂笼蛋白2(LCN2)是一种分泌蛋白,在脑缺血和神经退行性疾病中具有重要作用。塞络通胶囊(SLT)是由人参、银杏、藏红花三种中草药组成的治疗血管性痴呆的标准化制剂。虽然最近的临床试验已经证明SLT对血管性痴呆的有益作用,但其潜在的细胞机制尚未得到充分的探索。方法采用大鼠微球法致多发性脑梗死实验模型,通过Morris水迷宫实验评价SLT对恢复期大鼠空间记忆功能障碍的影响;随后,根据组织内及血清内炎性因子的表达,以及qPCR、免疫荧光、免疫印迹观察星形胶质细胞的活化、特异蛋白LCN2及其诱导的JAK2\STAT3信号通路的变化,以考察恢复期脑缺血损伤后星形胶质细胞的活化与炎性反应的关系及SLT对恢复期缺血损伤皮层区星形胶质细胞功能的影响;建立人星形胶质细胞糖氧诱导(OGD)损伤模型,并采用慢病毒转染的方法过表达目的基因LCN2,检测细胞分泌炎性因子含量变化,免疫荧光、蛋白免疫印迹观察星形胶质细胞的活化、特异蛋白LCN2及其诱导的JAK2/STAT3信号通路的变化,考察星形胶质细胞损伤后SLT有效成分对其的影响。结果 SLT可显著降低脑缺血引起的大鼠空间记忆认知功能障碍,减轻星形胶质细胞的活化及增殖,较为明显改善脑脑缺血损伤大鼠皮层缺血周边半暗带区神经元及突触超微结构损伤;SLT可抑制缺血侧皮层区及血清内炎性因子及趋化因子的表达,根据免疫荧光与免疫印迹的结果看出,SLT可能通过抑制LCN2诱导的JAK2/STAT3信号通路的表达来抑制炎性反应。结论 SLT介导神经炎症反应,通过LCN2-JAK2/STAT3通路抑制星形胶质细胞增生,抑制神经炎症反应,从而保护缺血性脑损伤,为缺血性脑卒中的治疗策略提供新的思路。     

神经病理性疼痛发生机制的研究进展

为了获得高效治疗神经病理性疼痛的方法,本研究对近几年国内外神经病理性疼痛机制与治疗方面的研究进行分析。大量动物模型实验研究证实,脊髓背角星形胶质细胞激活、C-纤维的敏化调节、嘌呤受体信号的激活以及TNF-α细胞因子释放等途径或机制共同作用导致神经病理性疼痛的发生。针对这些机制的研究可指导临床从不同途径阻断疼痛的发生及发展过程,从而治疗该类疼痛,并可为通过减少外周刺激、提高疼痛阈值、阻断疼痛感觉传导等相应手段治疗神经病理性疼痛提供重要理论依据。     

Chronic inflammatory pain and the neurovascular unit: a central role for glia in maintaining BBB integrity?

Pain is a complex phenomenon involving both a peripheral innate immune response and a CNS response as well as activation of the hypothalamic-pituitary-adrenal axis. The peripheral innate immune response to injury involves the rapid production and local release of proinflammatory cytokines such as tumor necrosis factor-alpha (TNF-/alpha), interleukin-1 (IL-1) and IL-6. Recent studies into the CNS response to peripheral chronic inflammatory pain strongly implicates a role for glia, and local synthesis of proinflammatory cytokines and growth factors. A characteristic feature of CNS inflammation is gliosis, in which inflammatory mediators activate glial cells (e.g. astrocytes and microglia, macrophages and leukocytes) which have been shown to induce and maintain hyperalgesia. In addition, inflammatory pain induces changes in blood-brain barrier (BBB) permeability and alters transport of clinically relevant drugs used to treat pain into the brain. Despite the increasing body of evidence for the involvement of glia in chronic pain and the role of glia in maintaining the BBB, few studies have addressed glial/endothelial interactions and the mechanisms by which glia may regulate the BBB during inflammatory pain. Further studies into the cellular mechanisms of glial/endothelial interactions may identify novel therapeutic targets for reversing chronic inflammatory induced BBB dysfunction and innovate therapies for modulating the severity of chronic inflammatory pain.     

比较研究奇正青鹏软膏与辣椒碱软膏的抗炎镇痛作用及机制

目的：探讨奇正青鹏软膏（简称青鹏软膏）和辣椒碱软膏在治疗关节炎疼痛作用及机制上存在的相似性及差异性。方法：分别用两种药物预处理大鼠,观察两药作用下福尔马林致痛模型大鼠的疼痛行为反应、局部组织病理变化、足底炎性痛局部组织P物质（SP）的表达、血清一氧化氮（NO）的水平以及血浆β-内啡肽（β-EP）的水平。结果：与模型组比较,青鹏软膏及辣椒碱各剂量组均能明显改善局部组织病理状况,并显著降低大鼠SP阳性细胞积分光度值（P〈0.01）;青鹏软膏两剂量组和辣椒碱大剂量组均抑制福尔马林II相疼痛、降低血清NO水平（P〈0.05）,并提高血浆β-EP的水平（P〈0.05）。与辣椒碱相比,青鹏软膏没有局部刺激作用,其作用具有剂量依赖性,并且对福尔马林Ⅰ相疼痛有抑制作用。结论：青鹏软膏与辣椒碱的镇痛作用机制有很大的相似性,但青鹏软膏的作用可能更加广泛和可控。     

cAMP/Epac信号通路调控慢性咳嗽及中药干预作用研究进展

慢性咳嗽属于临床疑难杂症,气道炎症、咳嗽神经通路敏感性增强以及炎性疼痛是其主要病理学基础。环磷酸腺苷(cyclic adenosine monophosphate,cAMP)/cAMP直接激活的交换蛋白分子(exchange protein activated by cAMP,Epac)信号通路广泛参与气道炎症(特别是气道神经源性炎症)、咳嗽神经兴奋性以及炎性疼痛等生理病理过程。本文就近年来cAMP/Epac信号通路调控气道炎症、咳嗽通路增敏、炎性疼痛而影响慢性咳嗽以及中药干预作用研究进行综述,为研究慢性咳嗽以及镇咳药的开发应用提供理论基础。     

Effect of synthetic cell-penetrating peptides on TrkA activity in PC12 cells

As TrkA, a high-affinity receptor of nerve growth factor (NGF), is a potential target for relieving uncontrolled inflammatory pain, an effective inhibitor of TrkA has been required for pain management. To identify a specific inhibitor of TrkA activity, we designed cell-penetrating peptides combined with amino-acid sequences in the activation loop of TrkA to antagonize tyrosine kinase activity. To select a peptide inhibiting TrkA activity, we examined the effect of cell-penetrating peptides on tyrosine kinase activity of recombinant TrkA in vitro and studied their effects on NGF-stimulated neurite outgrowth and protein phosphorylation in PC12 cells. Thereafter we investigated the effect of the selected peptide on NGF-stimulated TrkA activity and the expression of transient receptor potential channel 1 in PC12 cells. The selected peptide inhibited TrkA activity, but did not inhibit tyrosine kinase activities of other receptor-type tyrosine kinases in vitro. It also suppressed NGF-stimulated responses in PC12 cells. The selected synthetic cell-penetrating peptide antagonizing TrkA function would be a candidate for inflammatory pain therapy.     

Inhibition of Nav1.7 channel by a novel blocker QLS-81 for alleviation of neuropathic pain

Voltage-gated sodium channel Nav1.7 robustly expressed in peripheral nociceptive neurons has been considered as a therapeutic target for chronic pain, but there is no selective Nav1.7 inhibitor available for therapy of chronic pain. Ralfinamide has shown anti-nociceptive activity in animal models of inflammatory and neuropathic pain and is currently under phase III clinical trial for neuropathic pain. Based on ralfinamide, a novel small molecule (S)-2-((3-(4-((2-fluorobenzyl) oxy) phenyl) propyl) amino) propanamide (QLS-81) was synthesized. Here, we report the electrophysiological and pharmacodynamic characterization of QLS-81 as a Nav1.7 channel inhibitor with promising anti-nociceptive activity. In whole-cell recordings of HEK293 cells stably expressing Nav1.7, QLS-81 (IC₅₀ at 3.5 ± 1.5 μM) was ten-fold more potent than its parent compound ralfinamide (37.1 ± 2.9 μM) in inhibiting Nav1.7 current. QLS-81 inhibition on Nav1.7 current was use-dependent. Application of QLS-81 (10 μM) caused a hyperpolarizing shift of the fast and slow inactivation of Nav1.7 channel about 7.9 mV and 26.6 mV, respectively, and also slowed down the channel fast and slow inactivation recovery. In dissociated mouse DRG neurons, QLS-81 (10 μM) inhibited native Nav current and suppressed depolarizing current pulse-elicited neuronal firing. Administration of QLS-81 (2, 5, 10 mg· kg⁻¹· d⁻¹, i.p.) in mice for 10 days dose-dependently alleviated spinal nerve injury-induced neuropathic pain and formalin-induced inflammatory pain. In addition, QLS-81 (10 μM) did not significantly affect ECG in guinea pig heart ex vivo; and administration of QLS-81 (10, 20 mg/kg, i.p.) in mice had no significant effect on spontaneous locomotor activity. Taken together, our results demonstrate that QLS-81, as a novel Nav1.7 inhibitor, is efficacious on chronic pain in mice, and it may hold developmental potential for pain therapy.     

抗敏止痒乳膏的药效学

目的：考察抗敏止痒乳膏止痒镇痛，抗炎消肿的疗效。方法：通过两种疼痛模型，两种类症模型和一种致痒模型，用抗敏止痒乳膏进行实验性治疗和保护作用研究，并与新肤松进行了对比实验。结果：抗敏止痒乳膏具有显著的止痒、镇痛作用和良好的抗炎作用，并呈明显的量效关系。在相同剂量下，止痒、镇痛作用明显优于新肤松（P＜0．05或P＜0．01），抗炎作用与新肤松相似（P＞0．05），结论：证实了抗敏止痒乳膏治疗瘙痒性皮肤病的作用，为临床应用提供实验依据。     

Synthesis and Analgesic Effects of μ-TRTX-Hhn1b on Models of Inflammatory and Neuropathic Pain

μ-TRTX-Hhn1b (HNTX-IV) is a 35-amino acid peptide isolated from the venom of the spider, Ornithoctonus hainana. It inhibits voltage-gated sodium channel Nav1.7, which has been considered as a therapeutic target for pain. The goal of the present study is to elucidate the analgesic effects of synthetic μ-TRTX-Hhn1b on animal models of pain. The peptide was first synthesized and then successfully refolded/oxidized. The synthetic peptide had the same inhibitory effect on human Nav1.7 current transiently expressed in HEK 293 cells as the native toxin. Furthermore, the analgesic potentials of the synthetic peptide were examined on models of inflammatory pain and neuropathic pain. μ-TRTX-Hhn1b produced an efficient reversal of acute nociceptive pain in the abdominal constriction model, and significantly reduced the pain scores over the 40-min period in the formalin model. The efficiency of μ-TRTX-Hhn1b on both models was equivalent to that of morphine. In the spinal nerve model, the reversal effect of μ-TRTX-Hhn1b on allodynia was longer and higher than mexiletine. These results demonstrated that μ-TRTX-Hhn1b efficiently alleviated acute inflammatory pain and chronic neuropathic pain in animals and provided an attractive template for further clinical analgesic drug design.     

Brain cytokines as neuromodulators in cardiovascular control

1. The role of cytokines in cardiovascular control, especially in neurogenic hypertension, has received considerable attention during the past few years. Brain cytokines have been shown to exert profound effects on neuronal activity. Recently, a number of studies have shown that administration of pro‐inflammatory cytokines or anti‐inflammatory cytokines into the central nervous system has a significant impact on sympathetic outflow, arterial pressure and cardiac remodelling in experimental models of hypertension and heart failure. 2. Our objective in this review is to present a succinct account of the effect of cytokines on neuronal activity and their role in cardiovascular disease. Furthermore, we propose a hypothesis for a neuromodulatory role of cytokines in the neural control of cardiovascular function.     

康妇消炎栓联合盐酸莫西沙星治疗盆腔炎性包块的临床效果研究

目的：探讨康妇消炎栓联合盐酸莫西沙星治疗盆腔炎性包块的临床效果。方法选择本院2012年6月~2013年12月收治的盆腔炎性包块患者94例,将其随机分为对照组和研究组各47例,对照组患者给予盐酸莫西沙星治疗,研究组患者给予康妇消炎栓联合盐酸莫西沙星治疗,疗程为2周,比较两组的治疗效果。结果研究组治疗总有效率为95.7%,对照组治疗总有效率为78.7%,研究组治疗总有效率明显高于对照组,差异有统计学意义（P〈0.05）。治疗后短期内（1、3个月）两组盆腹部疼痛评分差异无统计学意义（P〉0.05）,治疗后6个月研究组盆腹部疼痛评分明显低于对照组,差异有统计学意义（P〈0.05）。结论康妇消炎栓联合盐酸莫西沙星治疗盆腔炎性包块能有效缓解患者的症状体征,效果确切,并且对盆腔炎主要后遗症---盆腹腔疼痛有较好的预防作用,值得临床推广应用。