CNQX对电刺激隐神经诱发大鼠扣带回前部神经元c-Fos基因表达的影响

目的阐明伤害性电刺激隐神经(saphenous nerve,SN)能否引起扣带回前部(anterior cingulategyrus,ACG)神经元c-Fos基因表达及其发生机制。方法用免疫组化方法研究伤害性电刺激SN后不同时间,ACG神经元c-Fos基因表达的变化,以及尾静脉注射α-氨基3-羟基5-甲基4-异恶唑丙酸/海人藻氨酸(α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid/kainate,AMPA/kainate)受体拮抗剂受体是谷氨酸受体之一,6-氰基-7-硝基喹喔啉-2,3-二酮(6-cyano-7-nitroquinoxaline-2,3-dike-tone,CNQX)对该变化的影响。结果伤害性电刺激SN后30 min ACG神经元Fos蛋白表达明显增加,60 min增加最明显,120 min后开始消退;并且尾静脉注射CNQX拮抗了伤害性电刺激SN引起的ACG神经元Fos蛋白表达的显著增加。结论伤害性电刺激SN能够引起ACG神经元Fos蛋白表达的显著增加,这种表达呈时间依赖性,提示ACG存有SN代表区,能够感受SN传入的伤害性信息;CNQX拮抗了伤害性电刺激SN引起的ACG神经元Fos蛋白表达的显著增加,提示AMPA/kainate受体参与此过程。     

大鼠鞘内注射AP5的抗内脏伤害作用

目的观察鞘内注射竞争性N-甲基-D-天冬氨酸（NMDA）受体拮抗剂2-氨基-5-磷酰基戊酸酯（AP5）对大鼠结直肠扩张（CRD）诱发的内脏伤害性刺激的影响及剂量关系。方法利用大鼠CRD模型诱导的内脏伤害性刺激,经鞘内引入药物,观察对腹壁撤回反射（AWR）达到3级时痛阈的影响。结果AP5剂量依赖地抑制CRD诱导的内脏伤害性刺激,并有时间作用特点。结论竞争性NMDA受体拮抗剂AP5可以抑制CRD诱导的内脏伤害性刺激,NMDA受体在内脏伤害性刺激脊髓水平的传递中起重要作用。     

GR82334对强电流刺激大鼠隐神经增强扣带回前部Fos蛋白表达的影响

目的研究尾静脉或蛛网膜下腔注射神经激肽(neurokinin,NK)-1受体拮抗剂GR82334对强电流刺激大鼠隐神经(saphenous nerve,SN)增强扣带回前部(anterior cingulate gyrus,ACG)Fos蛋白表达的影响。方法应用免疫组化技术进行实验研究。结果强电流刺激大鼠SN引起ACG Fos蛋白表达显著增强;尾静脉或蛛网膜下腔注射GR82334拮抗了强电流刺激大鼠SN引起的ACG Fos蛋白表达的显著增强。然而,蛛网膜下腔注射GR82334并没有完全拮抗强电流刺激大鼠SN引起的ACG Fos蛋白表达的显著增强。结论大鼠SN传导的伤害性信息能够到达ACG,激活c-fos基因表达;外周NK-1受体与中枢NK-1受体参与大鼠SN传入信息引起的ACG Fos蛋白表达增强的过程,但是,还存在其他递质和受体参与的大鼠SN信息传入的其它中枢通路引起ACG Fos蛋白表达的显著增强。     

中缝背核注射L-NAME抑制炎性痛大鼠脊髓Fos的表达

目的 观察中缝背核(DRN)内一氧化氮合酶(NOS)对炎性痛大鼠脊髓伤害性信息传递的调控作用.方法 建立大鼠单侧足底甲醛炎性痛模型,采用行为学、c-Fos免疫组织化学及烟酰胺腺嘌呤二核苷酸磷酸黄递酶(NADPH-d)组织化学技术,观察甲醛诱发炎性痛后大鼠DRN内NADPH-d、NADPH-d/Fos双标阳性神经元数量的变化;同时观察DRN内预先给予NOS抑制剂L-NAME对甲醛致痛大鼠疼痛评分及脊髓Fos蛋白表达的影响.结果 单侧足底注射甲醛后,DRN内NADPH-d、NADPH-d/Fos双标阳性神经元的数量增加.DRN内预先注射L-NAME降低炎性痛大鼠的疼痛学评分及脊髓Fos阳性神经元的数量.结论 DRN内NOS可能促进甲醛炎性痛大鼠脊髓伤害性信息的传递.     

NMDA和非NMDA受体参与介导猫脊髓内脏伤害性信息传递(英文)

目的:研究NMDA(N-methyl-D-aspartic acid)和非NMDA受体在介导脊髓内脏痛传入中的作用,方法:气球膨胀(3-15 kPa,20 s)麻醉猫结-直肠诱发脊髓背角痛敏神经元发放,结果:1)扩张结-直肠引起神经元发放增加的为兴奋性型:17个SLA型(短潜伏期突然增加);11个SLS型(短潜伏期渐增);9个LL型(长潜伏期),15个神经元属于抑制性的Inh型,2)67.6％,78.4％和59.5％的膨胀肠诱发兴奋的神经元,分别被微电泳NMDA、使君子酸(QA)和海人藻酸(KA)激活;60％,86.7％和53.3％的Inh神经元也分别被3个酸激活.3)微电泳NMDA受体拮抗剂d,l-2-amino-5-phosphonovalemte(APV)和非NMDA受体拮抗剂6,7-dinitro-quinoxaline-2,3-dione(DNQX),分别使兴奋性反应减少35％±10％和65％±14％,DNQX明显强于APV(P<0.05).DNQX使3/7个Inh神经元抑制翻转30％-50％,而APV无效,结论:NMDA和非NMDA受体均参与介导脊髓内脏伤害性信息传递,而非NMDA受体的作用更强。 (责任编辑 李颖)     

鞘内注射P物质拮抗氯胺酮的抗伤害作用

目的观察脊髓P物质对氯胺酮抗伤害作用的影响。方法在小鼠福尔马林实验中,结合行为学和Fos蛋白表达,观察鞘内注射(it)不同剂量的P物质对氯胺酮抗伤害作用的影响。结果氯胺酮20、30mg·kg-1ip可剂量依赖性地减少小鼠舔足时间(P<0.05)。P物质0.25、0.5ngit可增加注射氯胺酮小鼠舔足时间(P<0.05)。小鼠注射福尔马林后,注射侧脊髓背角Fos免疫样(Foslikeimmunoreactive,FLI)阳性神经元数量明显增加(P<0.01),预先给于氯胺酮30mg.kg-1ip可以明显减少脊髓背角FLI阳性神经元数量(P<0.01),而P物质0.5ngit能明显削弱氯胺酮对脊髓背角Fos表达的抑制(P<0.01)。结论鞘内注射P物质能拮抗脊髓水平氯胺酮抗伤害作用。     

异氟烷催眠、镇痛作用与NMDA受体甘氨酸位点的关系

目的分析异氟烷催眠、镇痛作用与NMDA受体甘氨酸位点的关系。方法建立小鼠异氟烷注射催眠、镇痛模型后,在小鼠催醒、甩尾、福尔马林实验中,观察侧脑室或鞘内注射NMDA受体甘氨酸位点的激动剂D-丝氨酸(D-Serine,D-Ser)后小鼠睡眠时间、甩尾潜伏期或累计舔足时间的变化;用免疫组化方法观察异氟烷及鞘内用药对福尔马林小鼠脊髓Fos蛋白表达的影响。结果侧脑室注射D-Ser对异氟烷的催眠时间无影响(P>0.05)。鞘内注射D-Ser(0.025、0.05、0.1ng)可拮抗甩尾实验、福尔马林实验Ⅰ相中异氟烷的镇痛作用(P<0.05,P<0.01),而对福尔马林实验Ⅱ相异氟烷的镇痛作用无影响(P>0.05)。鞘内注射D-Ser0.05ng可拮抗异氟烷对福尔马林小鼠脊髓Fos蛋白表达的抑制作用(P<0.01)。结论异氟烷催眠作用与脑内NMDA受体甘氨酸位点关系不大;脊髓NMDA受体甘氨酸位点介导异氟烷对热、化学刺激的镇痛作用,而与异氟烷对慢性炎性疼痛的镇痛作用无明显关系。     

鞘内注射哌唑嗪对氯胺酮抗伤害作用的影响

目的　探讨脊髓α1 受体和氯胺酮(Ket, 37. 5mg·kg-1,ip)抗伤害作用的关系。方法　用热水甩尾法观察大鼠鞘内预先注射α1 受体拮抗剂哌唑嗪(Pra, 10, 30μg)对Ket抗伤害作用的影响。并用c fos基因免疫组织化学技术,观察Ket对痛刺激诱发的大鼠脊髓c- fos表达的调节作用及鞘内预先注射Pra(30μg)对Ket调节作用的影响。结果　鞘内单独注射各剂量Pra对动物痛阈均无明显影响(P>0 .05), 鞘内预注Pra(10μg)对Ket抗伤害作用无明显影响(P>0 .05)。而鞘内预注Pra(30μg)则可明显减弱Ket抗伤害作用(P<0 .05)。痛刺激前给予Ket明显减少背角各层Fos免疫阳性神经元的数量(P<0 .05),Ket对痛刺激诱发的脊髓ⅠⅣ层c fos表达的抑制作用可被鞘内预注Pra所减弱(P<0 .05)。结论　脊髓α1 受体参与Ket抗伤害作用。     

NMDA受体NR2B亚单位在慢性内脏痛觉敏化中的作用

目的探讨外周与脊髓NMDA受体NR2B亚单位在慢性内脏痛觉敏化中的作用。方法模型组大鼠出生后d8～15,每天接受一次伤害性结直肠扩张刺激,8wk龄后用腹壁撤退反射(AWR)评估大鼠肠道敏感性。对腰骶背根神经节及胸腰与腰骶脊髓背角神经元进行免疫组织化学染色,比较对照与模型大鼠NR2B的表达。并比较对照与模型两组大鼠腹腔注射AP-7(NMDA受体拮抗剂)前后原发传入神经对结直肠扩张刺激的反应。结果①模型组大鼠AWR评分显著增高。②模型大鼠脊髓背根神经节NR2B亚单位表达增强。③模型大鼠脊髓内脏相关神经元NR2B亚单位表达增强。④AP-7显著抑制模型大鼠腰骶传入神经纤维对结直肠刺激的反应。结论NMDA受体NR2B亚单位可能参与慢性内脏痛外周与脊髓痛觉敏化的过程。     

P物质增强大鼠鞘内注射D—丝氨酸诱发的热痛过敏

目的：研究脊髓伤害性信息传递中P物质（SP）与N－甲基－D－天冬氨酸（NMDA）受体甘氨酸位点激动剂D－丝氨酸（D-serine)之间的功能联系。方法：在浅麻大鼠,采用行为学方法,测定甩尾反射潜伏期（TFL）并结合鞘内给药途径观察药物作用。结果：鞘内注射D-serine 1000nmol后1．5分钟,TEL明显缩短;在注射D-serine 10nmol前6分钟鞘内施加SP 0.05nmol,明显增强D-serine 10nmol引起的TEL缩短效应;选择性NMDA受体甘氨酸位点拮抗剂7－氯犬尿酸1pmol及非选择性PKC抑制剂H－7 10μmol均可阻断这种增强作用。结论：SP可使D－丝氨酸诱发的热痛过敏明显加强,NMDA受体甘氨酸位点及胞内蛋白激酶系统参与了脊髓SP与NMDA受体的相互作用。     

NO介导吗啡戒断大鼠脊髓Fos蛋白和NMDA_(1A)受体mRNA表达的增加

目的:观察NO在纳洛酮催促吗啡戒断大鼠脊髓神经元活动变化中的作用。方法:采用Fos免疫组织化学、NADPH-d组织化学、Fos/NADPH-d双标、鞘内注射、反义寡核苷酸和RT-RCR技术。结果:急性应用纳洛酮和慢性应用吗啡对大鼠脊髓Fos蛋白及NADPH-d阳性神经元表达无明显影响,二者也无Fos/NADPH-d双标神经元表达;纳洛酮催促吗啡戒断大鼠脊髓Fos蛋白、NADPH-d阳性神经元、纤维和终末表达明显增加,且出现Fos/NADPH-d双标神经元表达。预先鞘内注射nNOS反义寡核苷酸明显降低吗啡戒断症状评分,减少吗啡戒断大鼠脊髓Fos蛋白及NMDA_(1A)R mRNA表达。结论:NO介导吗啡戒断大鼠脊髓Fos和NMD_(1A)R mRNA表达的增加。     

四氢巴马汀同类物对福尔马林致痛诱导Fos蛋白表达的影响

目的 研究四氢巴马汀（ＴＨＰ）同类物对福尔马林致痛诱导的Ｆｏｓ蛋白表达的影响,以阐明ＴＨＰ同类物的镇痛机制。方法 在右后肢脚掌皮下注射５％福尔马林５０μＬ,诱发性疼痛,用免疫组织化学方法观察Ｆｏｓ的蛋白表达,结果：腹腔注射ＴＨＰ同类物和Ｄ２受体拮抗剂螺哌隆诱导的Ｆｏｓ蛋白表达主要位于纹状体和伏膈核,Ｄ２受体激动剂喹吡罗可阻滞ｌ－ＴＨＰ和螺哌隆诱导的Ｆｏｓ蛋白表达,ＴＨＰ同类物明显增加脑干下行痛觉调     

Activation of ERK／CREB pathway in spinal cord contributes to chronic constrictive injury-induced neuropathic pain in rats

Aim: To investigate whether activation and translocation of extracellular signalregulated kinase (ERK) is involved in the induction and maintenance of neuropathic pain, and effects of activation and translocation of ERK on expression of pCREB and Fos in the chronic neuropathic pain. Methods: Lumbar intrathecal catheters were chronically implanted in male Sprague-Dawley rats. The left sciatic nerve was loosely ligated proximal to the sciatica‘s trifurcation at approximately 1.0 mm intervals with 4-0 silk sutures. The mitogen-activated protein kinase kinase (MEK) inhibitor U0126 or phosphorothioate-modified antisense oligonucleotides (ODN) were intrathecally administered every 12 h, 1 d pre-chronic constriction injury (CCI) and 3 d post-CCI. Thermal and mechanical nociceptive thresholds were assessed with the paw withdrawal latency (PWL) to radiant heat and von Frey filaments. The expression of pERK, pCREB, and Fos were assessed by both Western blotting and immunohistochemical analysis. Results: Intrathecal injection of U0126 or ERK antisense ODN significantly attenuated CCI-induced mechanical allodynia and thermal hyperalgesia. CCI significantly increased the expression of p-ERK-IR neurons in the ipsilateral spinal dorsal horn to injury, not in the contralateral spinal dorsal horn. The time courses of pERK expression showed that the levels of both cytosol and nuclear pERK, but not total ERK, were increased at all points after CCI and reached a peak level on postoperative d 5. CCI also significantly increased the expression of pCREB and Fos. Phospho-CREB-positive neurons were distributed in all laminae of the bilateral spinal cord and Fos was expressed in laminae I and II of the ipsilateral spinal dorsal horn. Intrathecal injection of U0126 or ERK antisense ODN markedly suppressed the increase of CCI-induced pERK, pCREB and c-Fos expression in the spinal cord. Conclusion:The activation of ERK pathways contributes to neuropathic pain in CCI rats, and the function of pERK may partly be accomplished via the cAMP response element binding protein (CREB)-dependent gene expression.     

NMDA受体介导皮下注射福尔马林诱发的大鼠脊髓Fos蛋白的表达

AIM: To examine the effects of N-methyl-D-aspartate (NMDA) and non-NMDA receptors on noxious stimulation-induced Fos expression in the rat spinal cord. METHODS: Formalin (2%) was injected s.c. into one hindpaw of the rat. Fos expression was exhibited by immunocytochemical technique. RESULTS: Two hours after s.c. formalin, Fos-like immunoreactive (FLI) neurons were distributed mainly in medial part of the lamina I and the outer lamina II of the ipsilateral dorsal horn. dl-2-Amino-5-phosphonovalerate (APV) administered intrathecally (10 microL, 0.01, 0.1, or 1 g.L-1) before injection of formalin into a hindpaw reduced the number of FLI neurons dose-dependently in the dorsal horn (P < 0.01), while 6,7-dinitroquinoxaline-2, 3(1H,4H)-dione (DNQX) (1 g.L-1) was ineffective. CONCLUSION: NMDA receptor mediated noxious stimulation-induced Fos expression in the rat spinal cord.     

The spinal nitric oxide involved in the inhibitory effect of midazolam on morphine-induced analgesia tolerance

Previous studies had shown that pretreatment with midazolam inhibited morphine-induced tolerance and dependence. The present study was to investigate the role of spinal nitric oxide (NO) in the inhibitory effect of midazolam on the development of morphine-induced analgesia tolerance. Subcutaneous injection of 100 mg/kg morphine to mice caused an acute morphine-induced analgesia tolerance model. To develop chronic morphine tolerance in mice, morphine was injected for three consecutive days (10, 20, 50 mg/kg sc on Day 1, 2, 3, respectively). In order to develop chronic tolerance model in rats, 10 mg/kg of morphine was given twice daily at 12 h intervals for 10 days. Midazolam was intraperitoneally injected 30 min prior to administration of morphine. Tail-flick test, hot-plate and formalin test were conducted to assess the nociceptive response. Immunocytochemistry, histochemistry and western blot were performed to determine the effect of midazolam on formalin-induced expression of Fos protein, nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) and nitric oxide synthase (NOS) in chronic morphine-tolerant rats, respectively. The results showed that pretreatment with midazolam significantly inhibited the development of acute and chronic morphine tolerance in mice, which could be partially reversed by intrathecal injection of NO precursor L-arginine (L-Arg). In chronic morphine-tolerant rats, pretreatment with midazolam significantly decreased the formalin-induced expression of Fos and Fos/NADPH-d double-labeled neurons in the contralateral spinal cord and NADPH-d positive neurons in the bilateral spinal cord. Both inducible NOS (iNOS) and neuronal NOS (nNOS) protein levels in the spinal cord were significantly increased after injection of formalin, which could be inhibited by pretreatment with midazolam. The above results suggested that the decrease of the activity and expression of NOS contributed to the inhibitory effect of midazolam on the development of morphine tolerance.     

弗氏佐剂致炎大鼠脊髓背角Fos神经元的表达

目的探讨完全弗氏佐剂（CFA）致炎性痛大鼠Fos神经元（Fos-LIN）在脊髓背角痛觉传导通路中的表达及意义。方法①16只SD大鼠随机均分为两组,于右后肢踝关节外侧皮下分别注射0.9%生理盐水（NS）或CFA50μl,观察注射前后大鼠热缩足反射潜伏期TWL的变化;②48只大鼠随机平分为两组后分别注射NS和CFA（剂量和注射部位同前）以免疫组化法检测脊髓背角Fos-LIN表达。结果①大鼠注射CFA后TWL逐渐降低,12h时为最低点,持续3d后逐渐恢复,14d时基本恢复至基础水平;②NS组脊髓背角Fos-LIN散在出现,而CFA组中Fos-LIN1h时最先出现在大鼠右侧脊髓背角I-II层,4h时出现在V-VI层的数目逐渐增多,12h时为脊髓全层出现最多,至14d时在背角各层少见,且同组左侧脊髓各层Fos-LIN在1h-14d均比较少见。结论50μlCFA能诱导大鼠产生为期2周的炎性痛觉过敏。炎症期间脊髓背角Fos神经元的持续表达与外周痛觉信号的传导和中枢的调控有关。     

预先应用咪哒唑仑抑制小鼠和大鼠吗啡戒断反应

目的:研究咪哒唑仑对小鼠和大鼠吗啡戒断反应的影响.方法:实验中采用急性和慢性吗啡依赖和纳洛酮催促戒断模型.使用放免法测定cAMP含量,免疫组织化学方法观察Fos蛋白表达变化.结果:合用咪哒唑仑和吗啡可抑制小鼠急性和慢性吗啡依赖的发展.在急性吗啡依赖小鼠,咪哒唑仑-吗啡组纳络酮催促跳跃的ED_(50)(10.4,8.5-12.3 mg/kg)明显大于生理盐水-吗啡组(3.0,1.9-4.3 mg/kg)(P<0.01).在慢性吗啡依赖小鼠,咪哒唑仑-吗啡组纳络酮催促跳跃的发生率和跳跃次数明显低于生理盐水-吗啡组(P<0.01).预先使用咪哒唑仑抑制吗啡戒断大鼠脊髓Fos蛋白表达,但不能抑制脊髓cAMP含量的增加.结论:咪哒唑仑通过抑制脊髓神经元敏感化减轻吗啡戒断反应,cAMP信号转导通路不参与介导这一效应.