环氧化酶在神经病理性痛大鼠背根神经节P2X3受体表达上调中的作用

目的 探讨环氧化酶(COXs)在神经病理性痛大鼠背根神经节P2X3受体表达上调中的作用.方法 雄性SD大鼠24只,体重250～280g,采用随机数字表法,将其随机分为4组(n=6),神经病理性痛组(CCI组)、COX-1抑制剂组(Ⅰ组)和COX-2抑制剂组(C组)制备坐骨神经慢性缩窄性损伤(CCI)模型,假手术组(S组)仅暴露坐骨神经.于术后3～14 d Ⅰ组和C组分别以COX-1抑制剂布洛芬40mg·kg-1·d-1和COX-2抑制剂塞来昔布30mg·kg-1·d-1灌胃.分别于术前(基础状态)、术后3、5、7、10、14 d时测定热缩足潜伏期(PWL)和机械缩足阈值(PWT).然后处死大鼠,取L()-6节段背根神经节,测定P2X3受体mRNA及其蛋白表达水平.结果 与S组比较,CCI组术后PWL缩短,PWT降低,P2X3受体mRNA及其蛋白表达上调(P＜0.05);与CCI组比较,I组和C组术后PWL延长,PWT升高,P2X3受体mRNA及其蛋白表达下调(P＜0.05);与I组比较,C组术后PWL延长,PWT升高,背根神经节P2X3受体mRNA及其蛋白表达上调(P＜0.05).结论 COXs参与了神经病理性痛大鼠背根神经节P2X3受体表达上调,且COX-1的作用强于COX-2.

Abstract：

Objective To investigate the role of cyclooxygenases (COXs) in the up-regulation of the expression of P2X3 receptors in the dorsal root ganglion (DRG) in rats with neuropsthic pain. Methods Twenty-four male SD rats, weighing 250-280 g, were randomly divided into 4 groups ( n = 6 each): sham operation group (group S), chronic constrictive injury (CCI) group, COX-1 inhibitor ibuprofen group (group Ⅰ), and COX-2 inhibitor celecoxib group (group C). Neuropathic pain was induced by CCI. The animals were anesthetized with intraperitoneal 10% chloral hydrate 300-500 mg/kg. CCI was produced by placing 4 ligatures on the left sciatic nerve at 1 mm intervals. In group S, the left sciatic nerve was only exposed but not ligated. In groups Ⅰ and C, ibuprofen 40 mg·kg-1 ·d-1 and celecoxib 30 mg·kg-1 ·d-1 were given through a gastric tube into the stomach at day 3-14 after operation respectively. Paw withdrawal latency (PWL) and paw withdrawal threshold (PWT) were measured before operation (baseline), and at 3, 5, 7, 10 and 14 days after operation. Then the rats were sacrificed and their L()-6 DRGs were removed to detect the expression of P2X3 mRNA and protein. Results Compared with group S, PWL was significantly shortened, PWT decreased, and P2X3 mRNA and protein expression up-regulated in group CCI ( P ＜ 0.05＝. Compared with group CCI, PWL was significantly prolonged, PWT increased, and P2X3 mRNA and protein expression down-regulated in groups Ⅰ and C (P ＜0.05＝. Compared with group Ⅰ, PWL was significantly prolonged, PWT increased, and P2X3 mRNA and protein expression up-regulated in group C ( P ＜0.05＝. Conclusion COXs are involved in the up-regulation of the expression of P2X3 receptors in the DRG in rats with neuropathic pain, and the effect of COX-1 is stronger than that of COX-2.     

鞘内注射氯胺酮对神经病理性痛大鼠吗啡耐受时脊髓背角突触重塑的影响

目的 探讨鞘内注射氯胺酮对神经病理性痛大鼠吗啡耐受时脊髓背角突触重塑的影响.方法 鞘内置管成功的雄性SD大鼠48只,体重200～250 g,采用随机数字表法,将大鼠随机分为6组(n=8),置管后5 d,神经病理性痛组(NP组)、生理盐水对照组(NS组)、吗啡组(M组)、氯胺酮组(K组)和吗啡+氯胺酮组(MK组)采用背根神经节慢性压迫法制备神经病理性痛模型,假手术组(S组)仅暴露L5椎间孔.背根神经节慢性压迫后1 d NS组鞘内注射0.9%生理盐水20止,M组和K组分别给予吗啡20μg或氯胺酮50μg,MK组给予吗啡20μg+氯胺酮50μg,1次/d,连续14 d.分别于给药前(基础状态)、给药1、3、5、7、9、11、14 d时测定机械缩足阈值(PWT)和热缩足潜伏期(PWL).最后1d给药后立即处死大鼠,取脊髓组织,其中4只采用免疫组织化学方法测定脊髓背角突触数目,另外4只用于测定脊髓背角突触后膜致密物厚度.结果 与S组比较,其余5组PWT降低,PWL缩短,NP组、NS组、M组和K组突触数目和突触后膜致密物厚度增加(P＜0.05);与NP组比较,M组、K组和MK组PWT升高,PWL延长,突触数目和突触后膜致密物厚度降低(P＜0.05);与M组比较,MK组PWT升高,PWL延长,突触数目和突触后膜致密物厚度降低(P＜0.05).结论 鞘内注射氯胺酮可抑制神经病理性痛大鼠吗啡耐受时脊髓背角突触重塑.

Abstract：

Objective To investigate the effects of intrathecal (IT) ketamine on the synapsis remodeling in the spinal dorsal horn during devolopment of morphine tolerance in a rat model of neuropathic pain (NP). Methods Male SD rats weighing 200-250 g were used in this study. IT catheter was placed in the subarachnoid space according to Yaksh. Forty-eight SD rats in which IT catheter was successfully placed were randomly divided into 6groups (n=8 each): group sham operation (group S); group NP; group normal saline 20 μl IT(group NS);group morphine 20 μg IT (group M); group ketamine 50 μg IT (group K) and group morphine 20 μ g + ketamine 50 μg IT (group MK). NP was induced by compression of right L4,5 dorsal root ganglions with steel wire inserted through L4,5 intervertebral foramen in NP,M,K and MK groups. Normal saline or morphine and/or ketamine were injected IT once a day for consecutive 14 days. Paw withdrawal threshold (PWT) to mechanical stimulation and paw withdrawal latency (PWL) to a thermal nociceptive stimulus were measured on 0, 1, 3, 5, 7, 9, 11, 14 days during the consecutive 14 days of administration. The animals were sacrificed after the final IT administration. The lumbar segment of the spinal cord was removed for determination of the number of synapsis in the spinal dorsal horn by immuno-histochemistry in 4 animals in each group and observation of synaptic structure remodeling using electron microscope in another 4 animals in each group. Results Compared with group S, PWT was significantly decreased and PWL was shortened in the other 5 groups, and the number of synapsis was significantly increased and the synaptic structure was thickened in NP, NS, M and K groups (P ＜ 0.05). Compared with group NP,PWT was significantly increased and PWL was prolonged in M, K and MK groups, and the number of synapsis was significantly decreased and the thickness of synaptic structure was significantly reduced in group MK ( P ＜ 0.05).Compared with group M, PWT was significantly increased, PWL was prolonged, the number of synapsis was significantly decreased and the thickness of synaptic structure was significantly reduced in group MK ( P ＜ 0.05). Conclusion IT ketamine can inhibit the synaptic remodeling in the spinal dorsal horn during development of morphine tolerance in a rat model of NP.     

鞘内注射DREAM-shRNA对神经病理性痛大鼠脊髓背角p-CREB表达的影响

Objective To investigate the effects of intrathecal (IT) DREAM-short hairpin RNA (DREAM-shRNA) on expression of phosphorylated cyclic AMP response element binding protein (p-CREB) in the spinal dorsal horn in a rat model of neuropathic pain. Methods Adult male SD rats weighing 280-320 g were anesthetized with intraperitoneal 10% chloral hydrate. Neuropathic pain was induced by chronic constrictive injury (CCI) to sciatic nerve. IT catheters were placed according to the method described by Yaksh on 3rd day after CCI. Twenty-four rots in which IT catheter was successfully implanted were randomly divided into 4 groups (n = 6 each) : group Ⅰ sham operation (group S) ; group Ⅱ neuropathic pain (group NP) ; group Ⅲ RNA interference (group RNAi) and group Ⅳ blank vector (group BV). Lentivius with DREAM-shRNA 5 μl was injected IT in group RNAi, and blank vector 5 μl in group BV, and once a day for 7 days, starting from the day 8 after CCI. The mechanical pain threshold was measured at day 1 before CCI (T0 ,baseline) and day 7-14 after CCI (T1-8). The animals were killed on 15th day after CCI. The L4-6 lumbar segment of the spinal cord was removed for determination of the expression of green fluorescent protein (GFP) and p-CREB by immuno-fluorescent method.Results The mechanical pain threshold was significantly decreased as compared with the baseline at T0 in all 4 groups and returned to the baseline levels at T5-8 in group S and RNAi, but remained low in group NP and BY. The mechanical pain threshold was significantly lower after CCI/sham operation and significanty higher at T8 in group RNAi than in the other 3 groups. The expression of p-CREB in the spinal dorsal horn was up-regulated in group NP, RNAi and BV as compared with group S, and in group NP and BV as compared with group RNAi. The green fluorescence was observed in group RNAi but not in the other 3 groups. Conclusion IT DREAM-shRNA can ameliorate neuropathic pain in rats through inhibiting the expression of p-CREB in the spinal dorsal horn.     

侧脑室注射高选择性5-HT_（1A）受体拮抗剂或激动剂对大鼠神经病理性疼痛的影响

目的观察侧脑室注射高选择性5-HT1A受体激动剂8-OH-DPAT或拮抗剂WAY100635对大鼠神经病理性疼痛的影响。方法慢性结扎损伤（CCI）坐骨神经大鼠128只分为WAY100635组（W组,n=64）和8-OH-DPAT组（D组,n=64）,每组又分为8个亚组：WAY100635（1、2、4、8μg）-曲马多组（W1T组、W2T组、W4T组、W8T组）;WAY100635（1、2、4、8μg）-生理盐水组（W1N组、W2N组、W4N组、W8N组）;8-OH-DPAT（1、2、4、8μg）-曲马多组（D1T组、D2T组、D4T组、D8T组）;8-OH-DPAT（1、2、4、8μg）-生理盐水组（D1N组、D2N组、D4N组、D8N组）。另设空白对照组以同样方法暴露结扎坐骨神经（NN组,n=8）,建立CCI模型后腹腔和侧脑室均注射生理盐水。测定大鼠热痛敏阈（PWL）和机械痛敏阈（PWT）值。结果与NN组比较,W1T、W2T、W4T、W8T、W2N、W4N、W8N、D1T、D2T、D4T、D8T组PWL、PWT值明显升高（P〈0.01）。W4T、W8T、W2N、W4N、W8N组PWL、PWT值分别高于D4T、D8T、D2N、D4N、D8N组（P〈0.01）。结论曲马多的镇痛作用可以在中枢水平部分被5-HT1A受体激动剂拮抗,侧脑室注射5-HT1A受体拮抗剂WAY100635可以产生一定的镇痛作用。     

神经病理性痛大鼠背根神经节神经元γ氨基丁酸激活电流的改变

目的 探讨神经病理性痛大鼠背根神经节(DRG)神经元γ氨基丁酸(GABA)激活电流的改变.方法 健康成年SD大鼠20只,雌雄不拘,体重100～150g,采用随机数字表法,将大鼠随机分为2组,假手术组(S组,n=5),神经病理性痛组(NP组,n=15)行l3脊神经结扎诱发神经病理性痛.于术后5 d处死大鼠,S组急性分离L3-5DRG神经元,NP组急性分离L5DRG神经元,建立膜片钳全细胞记录模式.通过细胞外加药排管给予GABA 100μmol/L,记录各类型DRG神经元中GABA激活电流的发生率及电流幅度,记录给GABA前、后静息电位、动作电位(基强度、阈电位和超射值).结果 细胞外给予100μmol/L的GABA后可诱发部分神经元出现快速失活的内向电流.与给GABA前比较,S组给GABA后DRG大、中神经元出现明显去极化,静息电位升高,超射值和基强度降低(P＜0.05),阈电位差异无统计学意义(P＞0.05),NP组给GABA后DRG大、中神经元静息电位升高(P＜0.05),超射值、基强度和阈电位差异无统计学意义(P＞0.05).与S组比较,NP组DRG大、中神经元GABA激活电流的发生率和电流幅度均降低,静息电位、超射值、基强度的变化幅度降低(P＜0.05),阈电位的变化幅度差异无统计学意义(P＞0.05).S组和NP组部分DRG小神经元出现自发放电,具有自发放电的神经元上没有GABA激活电流出现.结论 神经病理性痛大鼠DRG大、中神经元上GABA介导的抑制信号的减弱,引起神经元兴奋性升高,可能是神经病理性痛的发病机制之一.

Abstract：

Objective To investigate the change in GABA receptor-activated current in dorsal root ganglion (DRG) neurons in rats with neuropathic pain. Methods Twenty adult SD rats of both sexes weighing 100-150 g were randomly divided into 2 gorups: sham operation group (group S, n = 5) and neuropathic pain group (group NP, n= 15). Neuropathic pain was induced by ligation of right L5 spinal nerve. The animals were sacrificed at 5 days after operation. The L5 DRG( neurons in group NP and L3-5 DRG neurons in group S were immediately isolated. Whole-cellpatch- clamp technique was used. The extracellular solution contained GABA 100μmol/L.The frequency and amplitude of the GABA-activated current in DRG neurons and the changes in action potential (threshold potential, rheobase and overshoot) and resting potential before and after GABA administration were recorded. Results GABA 100μmol/L induced rapid inactivation of inward current in most neurons. Compared with the baseline before application of GABA, in group S GABA induced depolarization,increased resting potential and decreased amplitude and rheobase of action potential in large and medium DRG neurons, while in group NP GABA increased resting potential but induced no significant change in threshold potential and rheobase and overshoot of action potential. The frequency and amplitude of GABA-activated current and the degree of change in resting potential and rheobase and overshoot of action potential were significantly lower in group NP than in group S.Spontaneous discharge occurred in small DRG neurons in both groups. No GABA-activated current was observed in all DRG neurons with spontaneous discharge. Conclusions Neuropathic pain is induced by decreasing GABA-mediated inhibition signals in large and medium DRG neurons leading to increased excitability of neurons.     

加巴喷丁对奥沙利铂诱发神经病理性痛小鼠背根神经节神经元高电压激活钙通道的影响

目的 评价加巴喷丁对奥沙利铂诱发神经病理性痛小鼠背根神经节(DRG)神经元高电压激活钙通道的影响.方法 清洁级雄性昆明小鼠,体重20～25 g,6周龄.采用腹腔注射奥沙利铂3mg/kg的方法制备神经病理性痛模型.取模型制备成功的小鼠41只,采用随机数字表法,将小鼠随机分为神经病理性痛组(NP组,n=20)和加巴喷丁组(G组,n=21),另取10只正常小鼠为正常对照组(C组).G组于奥沙利铂给药后第3天腹腔注射加巴喷丁100mg/kg,NP组及C组给予等容量的生理盐水,每天1次,连续3 d.于加巴喷丁给药前即刻、给药后1～3 d(T1-4)时测定小鼠双后肢机械缩足阈值(MWT).于最后一次MWT测定后,取两侧L4,5节段DRG,分离DRG神经元,采用全细胞膜片钳记录峰电流密度,拟合DRG神经元高电压激活钙通道激活曲线和稳态失活曲线,计算半数激活电位(Va1/2)和半数失活电位(Vi1/2).结果 与C组比较,NP组T1～4时、G组T1时MWT降低,NP组峰电流密度和Vi1/2升高(P＜0.05),Va1/2差异无统计学意义,G组峰电流密度、Vi1/2和Va1/2差异无统计学意义(P＞0.05);与NP组比较,G组T2-4时MWT升高,峰电流密度和Vi1/2降低(P＜0.05).结论 加巴喷丁减轻小鼠奥沙利铂诱发神经病理性痛的机制可能与抑制DRG神经元高电压激活钙通道电流,促进通道失活有关.

Abstract：

Objective To investigate the effects of gabapentin on high-voltage-activated calcium currents in dorsal root ganglion (DRG) neurons in mice with oxaliplatin-induced neuropathic pain (NP). Methods Pathogen-free male Kunming mice aged 6 weeks weighing 20-25 g were used in this study. NP was induced by injection of intraperitoneal oxaliplatin 3 mg/kg. Successful induction of NP was defined as the mechanical paw withdrawal threshold (MWT) measured at 3 d after oxaliplatin administration decreased to 40% of the baseline ( before administration of oxaliplatin). Forty-one mice in which NP was successfully induced were randomly divided into 2 groups: NP group ( n = 20) and gabapentin group (group G, n = 21 ). Another 10 normal mice served as control group (group C). At 3 days after oxaliplatin administration, gabapentin 100 mg/kg was injected intraperitoneally once a day for 3 consecutive days in group G, while C and NP groups received the equal volume of normal saline.MWT to von Fray filament stimulation was measured immediately before and 1-3 days after gabapentin administration (T1-4). After the last measurement of MWT, bilateral L4.5 DRG was collected and neurons were isolated. The high-voltage-activated calcium currents were recorded using whole-cell patch-clamp technique. The peak current density and the voltage where half of the current was activated ( Va1/2 ) or inactivated ( Vi 1/2 ) were calculated. Results Compared with group C, MWT at T1-4 was decreased, the peak current density and Vi1/2 were significantly increased in group NP, and MWT at T1 was decreased in group G ( P ＜ 0.05). There was no significant difference in the peak current density, Vi1/2 and Va1/2 between C and G groups ( P ＞ 0.05). MWT at T2-4 was significantly increased, while the peak current density and Vi1/2 were significantly decreased in group G compared with group NP (P ＜ 0.05). Conclusion Gabapentin can reduce oxaliplatin-induced NP in mice through inhibiting high-voltage-activated calcium currents and promoting the inactivation of the channels in DRG neurons.     

消炎镇痛液混合牛痘疫苗致炎兔皮提取物对大鼠坐骨神经损伤的效应

目的 评价消炎镇痛液混合牛痘疫苗致炎兔皮提取物(ERSVV)对大鼠坐骨神经损伤的效应.方法 成年SD大鼠50只,雌雄各半,体重260～300 g,采用随机数字表法,将大鼠随机分为5组(n=10):假手术组(S组)、坐骨神经慢性压迫性损伤组(CCI组)、消炎镇痛液组(A组)、EBSVV组(E组)和混合用药组(A+E组).CCI组、A组、E组和A+E组制备大鼠坐骨神经慢性压迫性损伤模型;S组仅暴露坐骨神经.在坐骨神经旁置管,于术后14 d开始给药,A组注射消炎镇痛液0.2Ml[地塞米松棕榈酸酯0.05ml(4 mg/ml)、2%利多卡因0.05 ml、维生素B120.05 ml(0.5 mg/ml)和注射用水0.05ml)];E组注射ERSVV 0.2ml(40 U/ml);A+E组注射消炎镇痛液(去掉注射用水0.05 ml)+ERSVV0.05 ml共0.2 ml.第1天给药后,地塞米松棕榈酸酯改为注射用水,持续给药14 d.分别于术前1 d(基础状态)、术后1、5、7、14、21、28 d时测定机械缩足阈值(MWT).5组分别于术后14、28 d时取5只大鼠,暴露坐骨神经,测定运动神经传导速度(NCV)及复合肌肉运动电位波幅(AP),然后取结扎处坐骨神经,光镜下观察病理学结果,计数轴突数,并测定髓鞘厚度.结果 与S组比较,CCI组MW、AP、轴突计数和髓鞘厚度降低,NCV减慢(P＜0.01);与CCI组比较,A组和E组MWT、AP、轴突计数和髓鞘厚度升高,NCV增快(P＜0.05);与A组或E组比较,A+E组MWT、AP、轴突计数和髓鞘厚度升高,NCV增快(P＜0.05或0.01).结论 与消炎镇痛液或ERSVV比较,两种药物混合应用可进一步减轻大鼠慢性压迫性坐骨神经损伤.

Abstract：

Objective To investigate the effect of anti-inflammation-analgesic injection (AIAI) combined with extract from rabbit skin inflamed by vaccinia virus (ERSVV) on repair of the damaged sciatic nerve in rats.Methods Fifty adult SD rats of both sexes weighing 260-300 g were randomly divided into 5 gronps ( n = 10each): sham operation group (group S); sciatic nerve chronic constriction injury group (group CCI); CCI + AIAI group (group A); CCI + ERSVV group (group E) and CCI + AIAI + ERSVV group (group A + E). Right sciatic nerve was exposed and 4 loose ligatures were placed on the sciatic nerve at 1 mm intervals with 4/0 catgut in CCI,A,E and A + E groups. A catheter was placed around sciatic nerve, and fixed to the nearby muscle and kept unclogged by injecting 0.2 nl distilled water daily. AIAI and/or ERSVV 0.2 ml were injected via the catheter starting from the 14th day after operation. AIAI 0.2 ml contained dexamethasone palmitate (4 mg/ml) 0.05 ml, 2% lidocaine 0.05 ml and vitamin B12 (0.5 mg/ml) 0.05 ml in distilled water. Dexamethasone palmitate was omitted in AIAI starting from the 2nd of drug administration in group A and A + E. Paw withdrawal threshold to mechanical stimulation (MWT) was measured before (baseline) and at 1, 5, 7, 14, 21 and 28 days after operation. Sciatic nerves were exposed at 14 and 28 days after operation in 5 rats in each group. Conduction velocity of motor nerve (NCV) and action potential (AP) of gastrocnemius muscle were measured. Sciatic nerve at the site of CCI was examined for pathologic changes, the number of axons (NA) and thickness of myelin sheath (TMS) with light microscope. Results CCI significantly decreased MWT, AP, NA, TMS and NCV in group CCI as compared with group S (P ＜0.01). AIAI and/or ERSVV significantly attenuated CCI-induced decrease in MWT, AP, NA, TMS and NCV in A,E and A + E groups as compared with CCI group ( P ＜ 0.05). Their curative effects were potentiated by combined use. Conclusions Both AIAI and ERSVV have curative effects against CCI-induced sciatic nerve injury and their actions are potentiated by combined used.     

神经病理性痛大鼠背根神经节TRESK mRNA表达的变化

目的 探讨神经病理性痛大鼠背根神经节(DRG) 孔钾离子通道TRESK mRNA表达的变化.方法 雄性SD大鼠32只,体重22、0～250 g,采用随机数字表法,将大鼠随机分为2组(n=16):假手术组(S组)和神经病理性痛组(NP组).采用坐骨神经分支选择性损伤法制备神经病理性痛模型.S组仅暴露神经,不结扎.于术前1 d和术后1、3、5、7、14 d取8只大鼠,测定左后肢机械缩足反应阈值(MWT)和热缩足潜伏期(TWL).于术前1 d和术后14 d痛阈测定结束后取L4,5术侧DRG,采用RT-PCR法测定TRESK mRNA的表达.结果 与S组比较,NP组MWT明显降低,DRG TRESK mRNA表达明显下调(P<0.05或0.01),TWL差异无统计学意义(P>0.05).结论 神经病理性痛大鼠DRG TRESK mRNA表达下调,该变化可能与神经病理性痛的形成有关.

Abstract：

Objective To evaluate the changes in the expression of diplopore potassium ion channel TRESK mRNA in dorsal root ganlion (DRG) in rats with neuropathic pain (NP) .Methods Thirty-two male SD rats weighing 220-250 g were randomly divided into 2 groups ( n = 16 each) : group sham operation (group S) and group NP. NP was induced by ligation and severance of left tibial and common fibular nerves according to the technique described by Decosterd. Eight rats in each group were sacrificed 1 day before and 14 day after operation and their L4,5 DRGs in the operated side were isolated for determination of TRESK mRNA expression by RT-PCR. In the remaining 8 rats in each group paw withdrawal threshold to mechanical stimuli ( MWT) and paw withdrawal latency to a thermal nociceptive stimulus (TWL) were measured at 1 day before (baseline) and 1, 3, 5, 7, 14 day after operation. Results MWT was significantly lower in group NP than in group S. The TRESK mRNA expression in L4,5 DRGs in the operated side was significantly decreased after operation as compared with the baseline before operation in group NP and was significantly lower in group NP than in group S. Conclusion The development and maintenance of NP may be closely related with down-regulation of TRESK mRNA.     

阿魏酸钠对婴幼儿体外循环血管内皮功能的保护作用

Objective Cardiopulmonary bypass (CPB) and its related ischemia reperfusion injury may cause endothelial cell injury.To study the protective effects of sodium ferulate in vascular endothelial function during CPB by testing the changes of vascular endothelial cell( CEC),nitric oxide( NO) and endothelin-1 ( ET-1 ) in children with congenital heart disease.Methods Sixty patients with congenital heart disease,including 28 males and 32 females were studied.The mean age was (19.7 ±10.4) months and body weight (10.5 ±6.1) kg.There were 37 VSD,8 ASD,7 TOF,5 TAPVC and 3 CAVC,among them 26 patients had pulmonary hypertension.They were randomly divided in to two groups:sodium ferulate group ( group S,n = 30),and control group ( group C,n =30) .Sodium ferulate (8 mg/kg) was given intravenously before CPB.Blood samples were taken from the arterial line at following time points:before CPB (TO),bypass 30 min(Tl ),the termination of CPB (T2 ),2h after operation ( T3 ) and 6h after operation ( T4 ),respectively for determination the concentration of vascular endothelial cell (CEC) in the blood,the concentration of nitric oxide (NO) and endothelin-1 ( ET-1) in the plasma.Results There were no significant difference for the two groups regarding above parameters at TO ( P > 0.05).The level of CEC was significantly elevated after CPB in both groups ( P < 0.05 ) .CEC were lower at T2 in group S than in group C ( P < 0.05 ) .NO was decreased in both groups,but was higher in group S at T2,T3 and T4 ( P < 0.05 ) .The concentration of plasma ET-1 was not significantly different before CPB,but there was a slight decrease at T1,and then it was significantly increased in both groups (P<0.05).But it was lower in group S than in group C at T1,T2,T3 and T4(P<0.05 orP<0.01).Conclusion There was severe endothelial cell damage during CPB.Sodium Ferulate can effectively antagonize the secretion of ET-1 to promote the formation of NO.Therefore,it reduces CPB-induced endothelial cell damage and protects vascular endothelial function during CPB.     

阿魏酸钠对婴幼儿体外循环血管内皮功能的保护作用

Objective Cardiopulmonary bypass (CPB) and its related ischemia reperfusion injury may cause endothelial cell injury.To study the protective effects of sodium ferulate in vascular endothelial function during CPB by testing the changes of vascular endothelial cell( CEC),nitric oxide( NO) and endothelin-1 ( ET-1 ) in children with congenital heart disease.Methods Sixty patients with congenital heart disease,including 28 males and 32 females were studied.The mean age was (19.7 ±10.4) months and body weight (10.5 ±6.1) kg.There were 37 VSD,8 ASD,7 TOF,5 TAPVC and 3 CAVC,among them 26 patients had pulmonary hypertension.They were randomly divided in to two groups:sodium ferulate group ( group S,n = 30),and control group ( group C,n =30) .Sodium ferulate (8 mg/kg) was given intravenously before CPB.Blood samples were taken from the arterial line at following time points:before CPB (TO),bypass 30 min(Tl ),the termination of CPB (T2 ),2h after operation ( T3 ) and 6h after operation ( T4 ),respectively for determination the concentration of vascular endothelial cell (CEC) in the blood,the concentration of nitric oxide (NO) and endothelin-1 ( ET-1) in the plasma.Results There were no significant difference for the two groups regarding above parameters at TO ( P > 0.05).The level of CEC was significantly elevated after CPB in both groups ( P < 0.05 ) .CEC were lower at T2 in group S than in group C ( P < 0.05 ) .NO was decreased in both groups,but was higher in group S at T2,T3 and T4 ( P < 0.05 ) .The concentration of plasma ET-1 was not significantly different before CPB,but there was a slight decrease at T1,and then it was significantly increased in both groups (P<0.05).But it was lower in group S than in group C at T1,T2,T3 and T4(P<0.05 orP<0.01).Conclusion There was severe endothelial cell damage during CPB.Sodium Ferulate can effectively antagonize the secretion of ET-1 to promote the formation of NO.Therefore,it reduces CPB-induced endothelial cell damage and protects vascular endothelial function during CPB.     

曲马多对神经病理性痛大鼠中脑远位触液神经元5-HT1A受体表达的影响

目的 探讨曲马多对神经病理性痛大鼠中脑远位触液神经元5-HT1A受体表达的影响.方法 SPF级雄性SD大鼠40只,体重220～280 g,采用坐骨神经慢性压迫法制备大鼠神经病理性痛模型,随机分为5组(n=8):正常对照组(C组)、生理盐水组(NS组)、曲马多组(T组)、神经病理性痛+生理盐水组(NP+NS组)和神经病理性痛+曲马多组(NP+T组).C组不行任何处理;NS组和T组仅暴露坐骨神经,分别腹腔注射生理盐水2 ml/kg或曲马多10 mg/kg;NP+NS组和NP+T组制备神经病理性痛模型,模型制备后第7天分别腹腔注射生理盐水2 ml/kg或曲马多10 mg/kg.除C组外,其余4组于腹腔注射曲马多或生理盐水前(T1)和注射后1 h(T2)时测定热痛阈和机械痛阈.于模型制备后第5天,左侧侧脑室注射30%霍乱毒素亚单位B与辣根过氧化物酶复合物(CB-HRP)3μl以标记远位触液神经元,并测定远位触液神经元5-HT1A表达水平.结果 与C组比较,NP+NS组中脑远位触液神经元5-HT1A受体表达下调(P＜0.05),其余组差异无统计学意义(P＞0.05);与NS组和T组比较,NP+NS组中脑远位触液神经元5-HT1A受体表达下调,热痛阈和机械痛阈降低,NP+T组热痛阈和机械痛阈降低(P＜0.05),中脑远位触液神经元5-HT1A受体表达差异无统计学意义(P＞0.05);与NP+NS组比较,NP+T组中脑远位触液神经元5-HT1A受体表达上调,热痛阈和机械痛阈升高(P＜0.05).结论 曲马多可下调中脑远位触液神经元5-HT1A受体的表达,该作用可能是其减轻大鼠神经病理性痛的机制之一.     

μ受体在抗神经生长因子抗体减轻大鼠骨癌痛中的作用

目的 评价μ受体在抗神经生长因子抗体(anti-NGF)减轻大鼠骨癌痛中的作用.方法 实验一健康雌性SD大鼠60只,体重200～220 g,随机分为4组(n=15):假手术组(S组)、假手术+anti-NGF组(SN组)、骨癌痛组(P组)和骨癌痛+anti-NGF组(PN组).P组和PN组于左侧胫骨上段骨髓腔内注射10μl Walker256乳腺癌细胞(1×105个)制备骨癌痛模型;S组和SN组于左侧胫骨上段注射PBS 10μl.于肿瘤细胞接种后13 d时,进行鞘内置管.鞘内置管成功后3 d,SN组和PN组鞘内注射anti-NGF 10μg(用生理盐水稀释至10μl),S组和P组鞘内注射生理盐水10μl,2次/d,连续5 d.于肿瘤细胞接种前、肿瘤细胞接种后13、16、18、21 d时测定自发缩足次数(NSF)、热缩足潜伏期(PWL)和机械性痛阈(PWT).肿瘤细胞接种后21 d时,处死大鼠,取L4.5段脊髓背角和背根神经节,测定μ受体及其mRNA的表达.实验二健康雌性SD大鼠30只,体重200～220 g,随机分为2组(n=15):骨癌痛+anti-NGF组(PN组)和骨癌痛+纳洛酮+anti-NGF组(PNN组).于左侧胫骨上段骨髓腔内注射10μlWalker256乳腺癌细胞(1×105个)制备骨癌痛模型.于肿瘤细胞接种后13 d时,进行鞘内置管.鞘内置管成功后3 d,PN组鞘内注射鞘内注入anti-NGF 10μg(生理盐水稀释至25μl);PNN组鞘内注射纳洛酮10μg(生理盐水稀释至25μl),0.5 h后,鞘内注射anti-NGF 10μg(生理盐水稀释至25 μl),2次/d,连续5 d.于肿瘤细胞接种前、肿瘤细胞接种后13、16、18、21 d时测定大鼠NSF、PWL和PWT.结果 实验一与S组比较,SN组NSF、PWL和PWT差异无统计学意义,SN组和PN组μ受体及其mRNA表达差异无统计学意义(P＞0.05),P组和PN组瘤细胞接种后13～21 d时NSF增加,PWL缩短,PWT降低,P组μ受体及其mRNA表达下调(P＜0.05或0.01);与P组比较,PN组肿瘤细胞接种后18～21 d时NSF减少,PWL延长,PWT升高,μ受体及其mRNA表达上调(P＜0.05或0.01).实验二与PN组比较,PNN组肿瘤细胞接种后18～21 d时NSF增加,PWL缩短,PWT降低(P＜0.05或0.01).结论 anti-NGF减轻大鼠骨癌痛与μ受体的激活有关.     

乳铁蛋白对神经病理性痛大鼠脊髓背角PKG活性的影响

目的 探讨乳铁蛋白对神经病理性痛大鼠脊髓背角cGMP依赖性蛋白激酶(PKG)活性的影响.方法 雄性SD大鼠32只,体重200～250 g,随机分为4组(n=8):假手术组仅分离坐骨神经,不结扎,鞘内注射生理盐水10μl+50%二甲基亚砜(DMSO)10μl;余3组采用结扎坐骨神经的方法制备大鼠神经病理性痛模型,神经病理性痛组鞘内注射生理盐水10μl+50%DMSO10μl;乳铁蛋白组鞘内注射乳铁蛋白100μg+50%DMS010μl;PKG抑制剂KT5823组鞘内注射乳铁蛋白100μg+KT582310μl.给药后180 min内每隔30 min以热刺激法测定大鼠缩爪潜伏期,随后处死大鼠取脊髓背角,采用免疫荧光法检测PKG活性,并行定量分析.结果 与神经病理性痛组和KT5823组相比,乳铁蛋白组缩爪潜伏期延长,乳铁蛋白组脊髓背角PKG活性升高(P＜0.05);神经病理性痛组与KT5823组上述指标比较差异无统计学意义(P＞0.05).结论 乳铁蛋白可通过抑制脊髓背角PKG活性减轻大鼠神经病理性痛.     

鞘内注射突触后致密物质-95反义寡核苷酸对神经病理性疼痛小鼠痛行为学的影响

目的 探讨鞘内给予突触后致密物质-95(postsynaptic density protein-95,PSD-95)反义寡核苷酸对坐骨神经结扎小鼠疼痛行为的影响. 方法 选取鞘内置管成功的C57BL/6雄性小鼠48只,采用随机数字表法分为4组(每组12只):假手术组(Sham组)、生理盐水组(NS组)、PSD-95反义寡核苷酸组(A组)、PSD-95错义寡核苷酸组(M组).采用结扎坐骨神经的方法制备小鼠坐骨神经慢性压迫性损伤(chronic constriction injury,CCI)模型,结扎坐骨神经后第1～14天,NS组、A组、M组分别鞘内注射生理盐水5μl、反义寡核苷酸5 μg/5μl、错义寡核苷酸5μg/5μl,1次/d.于术前1d及术后1、3、5、7、14、17、21 d测定小鼠结扎侧足底机械缩足反射阈值(paw withdrawal mechanical threshold,PWMT)和热缩足潜伏期(paw withdrawal thermal latency,PWL). 结果 A组小鼠术后1～14 d的疼痛阈值与Sham组维持一致[第14天,PWMT (5.69±1.34)g,P＞0.05;PWL(9.65±1.44)s,P＞0.05].术后17d,A组小鼠损伤侧足出现疼痛[PWMT (4.24±1.83)g,P＜0.05;PWL (7.18±0.41)s,P＜0.05],但是与NS组[PWMT (1.77±0.38)g,P＜0.05;PWL(4.33±1.21)s,P＜0.05]、M组[PWMT(1.6±0.37)g,P＜0.05;PWL (4.38±0.95)s,P＜0.05]比较,疼痛明显减轻,并且持续到术后第21天. 结论 在CCI所致神经病理性疼痛的发展阶段,连续鞘内给予PSD-95反义寡核苷酸可以完全逆转给药期内小鼠痛行为表现,并且在停止给药后7d仍有明显缓解疼痛的作用.     

杏仁核注射U0126对芬太尼诱发大鼠痛觉过敏的镇痛效应

目的探讨杏仁核中央核(Ce A)细胞外信号调节蛋白激酶(ERK)在芬太尼诱发痛觉过敏(OIH)发病机制中的作用。方法实验一:雄性SD大鼠12只,随机分为实验组(OIH模型组)和对照组,OIH建模成功后取右侧Ce A组织用Western blot检测p-ERK蛋白的表达水平。实验二:另取雄性SD大鼠30只,随机分为OIH组、OIH+U0124组、OIH+U0126(0.15 nmol)组、OIH+U0126(0.45 nmol)组和OIH+U0126(1.5 nmol)组,每组6只,均杏仁核置管后制作OIH模型,成功后向Ce A内分别注射0.3μl DMSO、U0124(1.5 nmol)、U0126(0.15、0.45、1.5 nmol);观测注药前后机械缩足阈值及热缩足潜伏期的变化。取Ce A组织检测p-ERK蛋白的表达水平。结果实验一:与对照组比较,实验组机械缩足阈值和热缩足潜伏期明显降低,Ce A区p-ERK蛋白表达明显升高(P0.05)。实验二:建模后各组大鼠机械缩足阈值及热缩足潜伏期均明显降低(P0.05),Ce A区pERK蛋白表达增加,U0126剂量依赖性地翻转上述行为学和分子水平的变化(P0.05)。结论Ce A区ERK参与了芬太尼诱发的痛觉过敏的维持,靶向抑制Ce A区ERK激活可治疗芬太尼诱发的痛觉过敏。     

神经病理性痛大鼠脊髓背角星形胶质细胞NF-κB活性的变化

目的 观察神经病理性痛大鼠脊髓背角星形胶质细胞NF-κB活性的变化,以探讨脊髓星形胶质细胞调控神经病理性痛时胞内可能的信号转导通路机制.方法 雄性SD大鼠16只,月龄2～3 71,体重220～280 g,随机分为2组(n=8):假手术组(S组)和神经病理性痛组(CCI组).CCI组采用慢性压迫性损伤法制备大鼠慢性神经病理性痛模型,S组仅暴露坐骨神经.分别于术前1 d和术后7 d测定机械痛阈和热痛阈,术后第7天测定痛阈后处死大鼠,取脊髓,记录腰段脊髓背角星形胶质细胞核内NF-κBp65的免疫反应阳性细胞数.结果 与术前1 d比较,CCI组大鼠术后7 d机械痛阈和热痛阈降低(P<0.05).与S组比较,CCI组大鼠术后7 d机械痛阈和热痛阈降低,术侧脊髓背角星形胶质细胞NF-κBp65免疫阳性细胞数增多(P<0.05).结论 脊髓背角星形胶质细胞参与大鼠神经病理性痛的调控,其机制可能与NF-κB信号转导通路有关.     

切口痛-瑞芬太尼痛觉过敏大鼠脊髓总蛋白及膜蛋白NMDA受体NRl 、NR2A及NR2B亚基表达的变化

目的 评价切口痛-瑞芬太尼痛觉过敏大鼠脊髓总蛋白及膜蛋白NMDA受体NR1、NR2A及NR2B亚基表达的变化.方法 尾静脉置管成功的雄性SD大鼠32只,体重240～260 g,月龄2～3月,采用随机数字表法,将大鼠随机分为4组(n=8):对照组(C组)静脉输注等容量生理盐水60 min,瑞芬太尼组(R组)静脉输注瑞芬太尼1.2μg·kg-1·min-1 60 min;切口痛组(I组)建立切口痛模型,同时静脉输注等容量生理盐水60 min;瑞芬太尼+切口痛组(R+I组)建立切口痛模型,同时静脉输注瑞芬太尼1.2 μg· kg-1 ·min-1 60 min.于生理盐水或瑞芬太尼给药前24h、给药后2、6、24和48 h时测定机械刺激缩足阈值(PWT)和热刺激缩足潜伏期(PWL),最后一次测定痛阈后处死取脊髓L4～6节段,采用Western blot法测定脊髓总蛋白及膜蛋白NMDA受体NR1、NR2A及NR2B亚基的表达,并计算膜蛋白中NR2B/NR2A比值.结果 与C组比较,I组、R组和R+I组PWT降低,PWL缩短,总蛋白及膜蛋白NMDA受体NR1和NR2B亚基表达上调,膜蛋白中NR2B/NR2A比值增加(P＜0.05).与R组和I组比较,R+I组PWT降低,PWL缩短,总蛋白及膜蛋白NMDA受体NR1和NR2B亚基表达上调,膜蛋白中NR2B/NR2A比值增加(P＜0.05).各组总蛋白及膜蛋白NMDA受体NR2A亚基表达差异无统计学意义(P＞0.05).结论 大鼠切口痛-瑞芬太尼痛觉过敏的形成可能与脊髓总蛋白及膜蛋白NMDA受体NR1和NR2B亚基表达上调和膜蛋白中NMDA受体NR2B亚基组成比例的增加有关.     

脊髓神经元型一氧化氮合酶在大鼠神经病理性痛中的作用

目的 探讨脊髓神经元型一氧化氮合酶(nNOS)在大鼠神经病理性痛中的作用.方法 健康雄性SD大鼠40只,体重220～280 g,采用结扎坐骨神经干的方法建立坐骨神经慢性压迫性损伤(CCI)模型.随机分为4组(n=10),Ⅰ组及Ⅱ组暴露坐骨神经干,分别于术后1 d开始鞘内注射选择性nNOS抑制剂7-NI 60 μg[溶于20%二甲基亚砜(DMSO)]10μl)、20%DMSO 10μl,1次/d,连续6d;Ⅲ组及Ⅳ组制备CCI模型,分别于术后1 d开始鞘内注射7-NI 60μg(溶于20%DMSO 10μl)、20%DMSO 10 μl,1次/d,连续6 d.分别于CCI前1 d、CCI后1、3、5、7 d时测定大鼠机械痛阈和热痛阈.于CCI后7 d,各组分别取5只大鼠,取术侧L_(4～6)背根神经节,分别采用实时定量PCR和Western blot法测定nNOS mRNA及蛋白的表达水平.结果 与Ⅰ组和Ⅱ组比较,T_(1～4)时Ⅲ组和Ⅳ组术侧后肢机械痛阈和热痛阈降低(P＜0.05),背根神经节nNOS蛋白及mRNA的表达上调(P＜0.05);与Ⅲ组比较,T_(1～4)时Ⅳ组机械痛阈和热痛阈降低,背根神经节nNOS蛋白及mRNA的表达上调(P＜0.05).结论 脊髓nNOS参与了大鼠神经病理性痛的形成.