不同感觉神经损伤对大鼠病理性疼痛的影响及其机制研究

目的 观察剪断大鼠的不同感觉神经分支引起的疼痛行为学表现及脑源性神经营养因子(BDNF)在其中的作用.方法 按随机数字表法将24只大鼠分为3组:SUR组,GS组和SHAM组,每组各8只,分别进行剪断Sural、GS神经和假手术.观察3组L5DRG和脊髓背角的BDNF表达情况及损伤的细胞类型.结果 GS组大鼠术后同侧后肢机械刺激50%缩足阈值较基础值和SHAM组显著下降(P＜0.01),热刺激撤足持续时间显著延长(P＜0.01),SUR组则无变化(P＞0.05).GS组L5DRG中BDNF阳性神经元比率[(37.87±4.23)%]和脊髓的BDNF阳性面积百分比[(21.9±3.1)%]较SHAM组增高[(分别为(17.31±2.12)%],(12.6±1.3)%],差异具有显著性(P＜0.01);而SUR组与SHAM组之间无统计学差异(P＞0.05).GS组表达BDNF的FG阳性神经元较SUR组明显增高(P＜0.01)[分别为(81.5%±3.8)%,(6.4±0.9)%];GS组中NF200与FG共染神经元占NF200阳性神经元的百分比较SUR组显著增高(P＜0.01)[分别为(47.7±1.8)%和(26.7±2.3)%].结论 切断骨骼肌的感觉神经会产生痛觉过敏,而切断皮肤的感觉神经则不会.不同类型神经元受损,以及DRG和脊髓背角的BDNF表达差异是导致这种痛觉差异的重要原因.

Abstract：

Objective To observe the pain behavioral performance of rats that different sensory nerve fibers were transected,and examine the expression of brain-derived neurotrophic factor(BDNF) in these models.Methods Twenty-four rats were divided into three groups according to random number table method:SUR group,GS group and SHAM group, which received sural nerve transection, gastrocnemius-soleus nerve transection or sham operation respectively.There were 8 rats in every group.The expression of BDNF in the lumbar 5 DRG and spinal dorsal horn were detected,and the types of damaged cells were also observed.Results In GS group, 50% paw-withdrawal thresholds were significantly decreased on the ipsilateral hind paw compared with baseline and with those in SHAM group,and the paw-withdrawal durations in response to the thermal stimulus increased significantly (P＜0.01 ＝.In contrast, no change was found in SUR group(P＞0.05 ).The expression of BDNF in the lumbar 5 DRG ( (37.87 ± 4.23 ) % ) and spinal dorsal horn ( (21.9 ± 3.1 ) % ) was significantly higher in GS group than in SHAM group( ( 17.31 ± 2.12 ) %, ( 12.6 ± 1.3 ) % ), and no significant difference was found between SUR and SHAM groups(P＞0.05 ).FG opposite cells which also expressed BDNF in GS group were more than those in SUR group ( (47.7 ± 1.8) % and (26.7 ± 2.3 ) % ) (P ＜ 0.01 ＝.The percentage in N200 and FG double positive cells to N200 positive cells in GS group was significantly increased in GS group than those in SUR group ( (47.7 ±1.8 ) %, (26.7 ± 2.3 ) % ) (P ＜ 0.01 ＝.Conclusion The data suggest that injury of the sensory nerve innervating skin does not produce hyperalgesia, but injury of the sensory nerve innervating muscle does.Different kinds of neuron were damaged and the differences of BDNF expression is essential for this difference.     

PKCε Mediates Substance P Inhibition of GABA_A Receptors-Mediated Current in Rat Dorsal Root Ganglion

The mechanism underlying the modulatory effect of substance P(SP) on GABA-activated response in rat dorsal root ganglion(DRG) neurons was investigated. In freshly dissociated rat DRG neurons, whole-cell patch-clamp technique was used to record GABA-activated current and sharp electrode intracellular recording technique was used to record GABA-induced membrane depolarization. Application of GABA(1–1000 μmol/L) induced an inward current in a concentration-dependent manner in 114 out of 127 DRG neurons(89.8 %) examined with whole-cell patch-clamp recordings. Bath application of GABA(1–1000 μmol/L) evoked a depolarizing response in 236 out of 257(91.8%) DRG neurons examined with intracellular recordings. Application of SP(0.001–1 μmol/L) suppressed the GABA-activated inward current and membrane depolarization. The inhibitory effects were concentration-dependent and could be blocked by the selective neurokinin 1(NK1) receptors antagonist spantide but not by L659187 and SR142801(1 μmol/L, n=7), selective antagonists of NK2 and NK3. The inhibitory effect of SP was significantly reduced by the calcium chelator BAPTA-AM, phospholipase C(PLC) inhibitor U73122, and PKC inhibitor chelerythrine, respectively. The PKA inhibitor H-89 did not affect the SP effect. Remarkably, the inhibitory effect of SP on GABA-activated current was nearly completely removed by a selective PKCε inhibitor epilon-V1-2 but not by safingol and LY333531, selective inhibitors of PKCα and PKCβ. Our results suggest that NK1 receptor mediates SP-induced inhibition of GABA-activated current and membrane depolarization by activating intracellular PLC-Ca2+-PKCε cascade. SP might regulate the excitability of peripheral nociceptors through inhibition of the "pre-synaptic inhibition" evoked by GABA, which may explain its role in pain and neurogenic inflammation.     

Differential expression of alpha-adrenoceptor subtypes in rat dorsal root ganglion after chronic constriction injury

Summary： mRNAs of alpha-adrenoceptor （α-AR） subtypes are found in neurons in dorsal root ganglion （DRG） and change after peripheral nerve injury. In this study, the distribution of α-AR subtype proteins was studied in L5 DRG of normal rats and rats with chronic constriction injury of sciatic nerve （CCI）. Using immunofluorescence technique, it was found that α1A-, α1B-, and α2A-AR proteins were expressed in large, medium, and small size neurons in normal DRG, and significantly increased in all size neurons 14 days after CCI. α1D- and α2C-AR was also expressed in all size neurons in normal DRG. However, α1D-AR was significantly increased and α2C-AR was decreased in small size neurons 14 days post CCI. α2B-AR neurons were not detectable in normal and CCI DRG. Co-expression of α1A- and α2A-AR in the same neuron was observed in normal DRG and increased post CCI. Collectively, these results indicated that there is distinct distribution of α-AR subtypes in DRG neurons, and the distribution and levels of expression of α-AR subtypes change differently after CCI. The up-regulation of α-AR subtypes in DRG neurons may play an important role in the process of generating and transmitting neuropathic pain.     

Different suppressive effect of lidocaine on persistent Na^＋ current and transient Na^＋ current in dorsal root ganglion neurons

Objective ：To investigate the different suppressive effect of lidocaine on persistent Na^＋ current and transient Na^＋ current in injured or uninjured dorsal root ganglion neurons. Methods： Totally 23 SD rats were randomly divided into 2 groups： control group （n： 10） and chronically compressed DRG （dorsal root ganglion） group （CCD group, n= 13）. Rats were anesthetized and DRG was isolated. Single DRG neuron was isolated by enzymatic disassociation method. Persistent Na^＋ current （INap） and transient Na^＋ current （INaT） were elicited in voltage clamp mode. Results： The presence of INap was testified in most DRG neurons （38/46 neurons in CCD group and 31/39 neurons in control group, P〉0. 05）. However, the cur- rent density of INap in CCD group （4. 6±0. 6 pA/pF, n=38 neurons） was greater than that in control group （2.5±0.4 pA/pF, n=31 neurons） （P〈0. 05）. The characteristics of INap was observed and found that INap could he blocked by 0.2 μmol/L tetrodotoxin easily. Furthermore, the does-effect relationship of lidocaine on INaP and IN.T were also examined. INaP and IN.T were suppressed by different concentrations of li- docaine, the range for INap was 5-20 μmol/L and for INaT was 0. 05-2 mmol/L. Conclusion： INap and INaT were suppressed by different concentrations of lidocaine. INap was suppressed by very low concentration of lidocaine （5-20 μmol/L）. However, INaT could only be blocked by high concentration of lidocaine （0.05-2 mmol/L）.     

慢性脑低灌注大鼠海马神经元损伤的机制研究

目的 探究慢性脑低灌注大鼠海马神经元损伤的机制.方法 双侧颈总动脉结扎(2VO)制备慢性脑低灌注大鼠模型,2VO术后8周取材,分别进行HE染色、电镜、流式细胞术以及Western Blotting检测.结果 HE结果显示2VO组的海马神经元排列稍紊乱,且数量与假手术对照相比有不同程度减少[CA2区:(34.75±3.40)个,(49.25±9.67)个,P＜0.05;DG区:(73.50±9.26)个,(90.75±4.35)个,P＜0.05];电镜观察发现2VO组大鼠海马区部分神经元胞核中略有固缩,出现异染色质边集,胞浆则出现水肿,以线粒体与内质网尤甚;流式细胞术结果表明2VO组海马神经元早期凋亡率[(9.117±2.540)%]高于假手术对照组[(4.750±3.481)%],差异有统计学意义(P＜0.05);Western Blotting检测发现procaspase-3在2VO组大鼠海马的表达与假手术对照组差异无显著性(P＞0.05).结论 慢性脑低灌注大鼠海马神经元的损伤主要由凋亡造成.

Abstract：

Objective To explore the the cellular damage of hippocampus neuron in a rat model of chronic cerebral hypoperfusion. Methods Rat model of chronic cerebral hypoperfusion was established by permanent bilateral common carotid arteries occlusion (2VO). Eight weeks after the operation,the brains were removed and examined with histological stains, electron microscope, flow cytometer and Western Blotting. Results Compared with the control group,the arrangement of hippocampus neurons in 2VO rats appeared to be more irregular, and the number of the neurons decreased partly ( CA2: ( 34.75 ± 3.40) vs (49.25 ± 9.67 ), P ＜ 0. 05; DG: ( 73.50 ± 9.26)vs ( 90.75 ± 4.35 ), P ＜ 0. 05 ). By electron microscopic study of hippocampus neurons in 2VO rats, the nuclei became smaller and the heterochromatin assembled in the border of the nuclei in some neurons, while cytoplasm swelled,especially in mitochondria and endoplasmic reticulum. The rate of apoptosis of hippocampus neurons in 2VO rats( (9. 117 ±2. 540)% ) ,detected by the flow cytometer,was higher than that of sham group( (4. 750 ±3.481 ) % ) (P ＜ 0. 05 ). The expression of pro-caspase-3 in hippocampus of 2 VO rats was not altered significantly compared with the control group(P ＞ 0. 05 ). Conclusion The cellular damage of hippocampus neuron in 2VO rats was mainly caused by apoptosis.     

加巴喷丁对SNL模型大鼠损伤DRG神经元细胞内[Ca2+]i的影响

目的:探讨加巴喷丁对脊神经结扎(SNL)模型大鼠损伤背根神经节神经元胞内游离钙浓度的影响?方法:SD大鼠,雄性35只,4~6周龄,采用左侧L5 SNL术制备大鼠神经病理性疼痛模型?于结扎后15 d采用酶消化法急性分离损伤同侧L5背根神经节神经元(SNL组)和假手术组L5背根神经节神经元(Sham组)?采用流式细胞仪检测10?100和300 μmol/L浓度加巴喷丁对损伤侧L5背根神经节神经元基础及高K+激发胞内游离钙浓度的影响?结果:加巴喷丁剂量依赖性地抑制高钾激发的胞内游离钙浓度的增加(P < 0.05或P < 0.01),但对静息钙无作用?结论:加巴喷丁对神经病理性疼痛大鼠损伤背根神经节神经元高钾激发的钙内流的抑制作用,可能与其抗伤害作用的外周机制相关?     

Downregulation of metabotropic glutamate receptors mGluR5 and glutamate transporter EAAC1 in the myenteric plexus of the diabetic rat ileum

Objective: To study the morphologic abnormalities of the myenteric plexus in diabetic rats and to explore the mechanism of their effect on gastrointestinal motility. Methods: Forty rats were randomly divided into a diabetic group and a control group, Gastric emptying and small intestine transit rates were measured and histologic and molecular changes in glutamatergic nerves in the ileal myenteric plexus were observed, mGluR5 receptor and EAAC1 transporter changes in the diabetic rats were studied using fluorescence immunohistochemistry and RT-PCR. Results:Eighteen weeks after the establishment of the diabetic rats model, gastric emptying and small intestine transit rates were found to be significantly delayed in the diabetic group when compared with the control group. The density of glutamatergic ganglia and neurons in the ileal myenterie plexus were significantly decreased in the diabetic group when compared with control group(P < 0.05) and the mGluR5 receptors and EAAC1 transporters were downregulated in the diabetic rats(P < 0.05). Conclusion: Decreased glutamatergic enteric ganglia and neurons and decreased mGluR5 receptors and EAAC1 transporters in the intestinal myenteric plexus is one of the mechanisms of diabetic gastroenteropathy in rats.     

Influence of Yanyankang Powder(眼炎康散)on Th1/Th2 in Rats with Experimental Autoimmune Uveitis

Objective:To study the influence of Yanyankang Powder(眼炎康散) on Th1/Th2 in rats with experimental autoimmune uveitis(EAU).Methods:The EAU models were induced in Lewis rats by immunization with interphotoreceptor retinoid binding protein(IRBP) 1177-1191 in complete Freund's adjuvant.The rats were randomly divided into 3 groups:a model control group,a Yanyankang group,and a prednisone group,9rats in each group.The model control group was intervened with saline solution by gavage.The Yanyankang group was intervened with Yanyankang Powder 4 g/(kg·day) by gavage.The prednisone group were intervened with prednisone acetate tablets 5 mg/(kg·d) by gavage.All groups were intervened after immunization once every 2 days for 18 days and monitored by slit-lamp biomicroscopy daily until day 18.The levels of gamma interferon(INF-7) and interleukin-10(IL-10) in the supernatants of T cells were determined by enzyme-linked immunosorbent assay.Polymerase chain reaction(PCR) technology was used for measuring Th1 and Th2 related cytokine mRNA expressions.Results:Slighter intraocular inflammation was found in the Yanyankang group and the prednisone group than the control group.The levels of the IFN-γ and IL-10 in the supernatants of the spleen lymph node cells were 382.33 ± 6.30,155.87 ±4.46 μ/L in the Yanyankang group and270.93 ± 7.76,265.32 ±11.88 μg/L in the prednisone group.Both had significant differences compared with the control group(941.53 ± 8.59,20.67 ±4.65 μg/L;P=0.01).The PCR results showed the same tendency.Conclusion:Yanyankang Powder showed favorable effects in the rats with EAU by influencing the function of Th1 and Th2 cells.     

Expression changes of parvalbumin and microtubule-associated protein 2 induced by chronic constriction injury in rat dorsal root ganglia

Background  Parvalbumin (PV), as a mobile endogenous calcium buffer, plays an important role in affecting temporospatial characteristics of calcium transients and in modulating calcium homeostasis. PV is expressed in neurons in the dorsal root ganglion (DRG) and spinal dorsal horn and may be involved in synaptic transmission through regulating cytoplasm calcium concentrations. But the exact role of PV in peripheral sensory neurons remains unknown. Microtubule-associated protein 2 (MAP-2), belonging to structural microtubule-associated protein family, is especially vulnerable to acute central nervous system (CNS) injury, and there will be rapid loss of MAP-2 at the injury site. The present study investigated the changes of PV expressing neurons and the MAP-2 neurons in the DRG after an operation for chronic constriction injury to the unilateral sciatic nerve (CCI-SN), in order to demonstrate the possible roles of PV and MAP-2 in transmission and modulation of peripheral nociceptive information.

Methods  Seventy-two adult male Sprague-Dawley (SD) rats, weighing 180–220 g, were randomly divided into two groups (36 rats in each group), the sham operation group and chronic constriction injury (CCI) group. Six rats in each group were randomly selected to receive mechanical and thermal sensitivity tests at one day before operation and 1, 3, 5, 7, and 14 days after surgery. After pain behavioral test, ipsilateral lumbar fifth DRGs were removed and double immunofluorescence staining was performed to assess the expression changes of PV and of MAP2 expressing neurons in the L5 DRG before or after surgery.

Results  The animals with CCI-SN showed obvious mechanical allodynia and thermal hyperalgesia (P <0.05). Both the thermal and mechanical hyperalgesia decreased to their lowest degree at 7 days after surgery compared to the baseline before surgery (P <0.01). In normal rats before surgery, a large number of neurons were MAP-2 single labeled cells, and just a small number of PV-expressed neurons were found. PV-positive neurons, PV-positive nerve fibers and PV-negative neurons, formed a direct or close contact for cross-talk. We used immunocytochemical staining to quantify the time course of changes to PV and MAP-2 expressing neurons in tissue, and found that the number of PV expressing neurons began to slightly decrease at 3 days after surgery, and had a significant reduction at CCI day 5, day 7 (P <0.05). But MAP-2 neurons significantly decreased on just the 3rd day after CCI (P <0.05). No changes in PV and MAP-2 expression were almost found in sham operated rats. The number of PV positive neurons, was positively correlated with the hyperalgesia threshold.

Conclusions  A sharp decline in MAP-2 neurons may be the early response to surgical injury, and PV positive neurons were much more effective at affecting the changes of pain behaviors, indicating that the down-regulation of PV protein could participate in, at least in part, the modulation of nociceptive transmission.

     

丁基苯酞对慢性脑缺血大鼠脑组织caspase-3表达的影响

目的 观察丁基苯酞(NBP)对慢性脑缺血大鼠脑组织中caspase-3表达的影响.方法 将大鼠随机分成对照组、模型组和治疗组,采用双侧颈总动脉结扎法建立大鼠慢性脑缺血模型,HE染色观察神经元形态学改变,免疫组化法观察各组大鼠大脑皮层和海马中caspase-3表达情况.结果 慢性脑缺血后,大鼠脑组织中caspase-3表达水平增高,经NBP治疗后,表达水平降低(P<0.05).结论 NBP可以通过下调慢性脑缺血组织中caspase-3的表达发挥神经保护作用. Abstract： Objective To observe the effect of dl-butylphthalide(NBP) on the expression of caspase-3 in cortex and hippocampus after chronic cerebral ischemia in rats. Methods Chronic cerebral ischemia models were established by the permanent ligation of bilateral common carotid arteries. Sixty rats were randomly divided into control group, model group and NBP-treated group. The morphologic change of the neurons was observed with HE stainingand the expression of caspase-3 in rat brain cortex and hippocampus was measured by immunohistochemical technique. Results The expression of caspase-3 in cortex and hippocampus was significantly increased in model group compared with that in control group, but decreased in NBP-treated group compared with that in model group(P<0.05). Conclusions NBP may exert neuroprotective effect by decreasing the expression of caspase-3 in cortex and hippocampus after chronic cerebral ischemia in rats.     

外周神经损伤后对大鼠脊髓运动神经元及背根节感觉神经元的影响

大鼠左腓总神经离断后与胫神经端侧吻合,于术后第1至8周取L4～5脊髓和背根节（DRG）做冰冻切片,分别显示前角运动神经元（SMN）AChE活性和DRG神经元NOS活性。结果发现;①正常鼠SMN－AChE活性中－强度阳性反应。实验组外侧核SMN－AChE活性和阳性细胞数,于术后2～3周比正常者明显减低,有的SMN－核周体出现空泡样变性;术后4～8周,SMN酶活性和阳性细胞数渐增,并明显高于正常者。同体对照组与实验组同期比较,SMN酶活性相对较弱。②正常鼠部分中－小型DRG神经元NOS活性为中－强度阳性反应。实验组于术后第1～4周,NOS活性和阳性细胞数较正常者明显增高;术后第5～8周其渐趋正常。对照组DRG神经元NOS活性表达较强及与实验组有相应的变化梯度。二组DRG内均见到空泡样变性的神经元核周体。结果提示,坐骨神经慢性损伤所产生SMN－AChE活性增强,可能是轴突再生的反应,并对由NO介导的一级感觉信息传导有一定影响。     

天麻素对大鼠离体胸主动脉血管环的作用及机制研究

目的：观察天麻素对大鼠离体胸主动脉环的作用,并探讨其作用机制。方法采用离体动脉环灌流方法,观察累加浓度天麻素对大鼠离体胸主动脉环的直接作用;同法观察不同浓度天麻素(5μmol/L、50μmol/L、100μmol/L、150μmol/L、200μmol/L、250μmol/L)对10-6 mol/L去氧肾上腺素(PE)预收缩有内皮和去内皮血管环的作用;用无Ca2+K-H液和不同浓度天麻素孵育对PE收缩去内皮血管环的作用;观察200μmol/L天麻素对60 mmol/L KCl预收缩血管环的作用。结果天麻素直接作用于血管环时,天麻素组与对照组血管紧张度比较差异无统计学意义(P>0.05);天麻素(≥50μmol/L)作用于PE预收缩血管环时,有内皮天麻素组血管环舒张率分别高于有内皮对照组[(41.9±8.9）%、(57.6±9.1)%、(65.0±10.2)%、(75.6±11.7)%、(81.9±12.0)%vs (10.5±1.6)%、(19.4±5.9)%、(22.6±7.2)%、(24.9±6.8)%、(25.1±7.3)%、P<0.05],去内皮天麻素组血管环舒张率分别高于去内皮对照组[(20.6±2.9)%、(36.5±8.2)%、(43.7±9.3)%、(55.7±9.6)%、(58.7±9.6)%vs (9.8±2.1)%、(15.8±4.7)%、(18.6±6.5)%、(20.3±5.0)%、(21.5±5.1)%,P<0.05],且均有浓度依赖性,多组间比较F值分别为5.67,5.99,6.11,6.47,7.05,P<0.05;天麻素作用于无钙K-H液中PE收缩去内皮血管环时,实验组舒张率[(5.1±1.2)%]小于对照组[(12.8±3.7)%],差异有统计学意义(t=5.599,P<0.05)。天麻素处理作用于60 mmol/L KCl预收缩的血管环时,天麻素组[（96.7±8.6）%]与对照组[(98.6±7.9)%]张力变化差异无统计学意义(t=0.581,P>0.05)。结论天麻素对正常血管无舒张作用;天麻素能舒张PE预收缩的血管,其机制是通过抑制血管平滑肌IP3受体介导的内钙释放;它不能抑制α1受体依赖性钙通道和电压依赖性钙通道;天麻素舒张PE收缩的血管的作用有内皮依赖性和浓度剂量依赖效应。     

依达拉奉通过降低脊神经结扎大鼠背根神经节和脊髓pJNK抑制痛敏

目的:观察依达拉奉对脊神经结扎(SNL)大鼠痛敏的影响及其作用机制。方法:将SD雄性大鼠随机分为假手术(Sham)组、SNL模型组和依达拉奉(Eda)组,观察脊神经结扎大鼠术前及术后7 d机械痛反应阈值(PWMT)的变化;在术后相应时间点取各组大鼠手术侧L5和L4及对侧L5背根神经节(DRG)和脊髓,观察pJNK在DRG和脊髓中的表达分布及依达拉奉对pJNK表达的影响。结果:依达拉奉可以减轻脊神经结扎引起的痛敏现象;SNL组术后24 h手术侧L5DRG中pJNK阳性神经元的表达增加,术后3 d免疫双荧光显示脊髓中 pJNK在星型胶质细胞中存在高表达现象;依达拉奉可以降低相应时间点DRG和脊髓中pJNK表达的含量。结论:依达拉奉能抑制脊神经结扎引起的痛敏现象,其机制可能是通过降低脊髓和DRG中pJNK的含量而产生镇痛作用的。     

NGF和GM1联合应用对坐骨神经损伤大鼠初级传入神经元的保护作用

目的:探讨联合应用神经营养因子(NGF)和神经节苷脂（GM1）对大鼠周围神经损伤后初级传入神经元的保护作用。方法: 选用SD大鼠96只，随机分为对照组、NGF组、GM1组和NGF + GM1组。将大鼠右侧坐骨神经切断，神经两断端相距5?mm，硅胶管桥接，用不同的药物处理后，于术后不同时间取出背根神经节进行研究。结果：4周时神经元数和神经传导速度，NGF+GM1组多于或快于其它各组(P＜0.01），NGF组和GM1组均多于或快于对照组(P ＜0.01）；8周时，各实验组多于或快于对照组(P＜0.01），各实验组之间差异无统计学意义（P＞0.05）。结论:周围神经损伤后，联合应用NGF和GM1对初级传入神经元的保护作用优于单用NGF、GM1，且起效早。     

外源性表皮生长因子对大鼠坐骨神经损伤后感觉神经元超微结构变化的影响

目的应用外源性表皮生长因子(EGF)观察神经损伤后对背根神经节感觉神经元超微结构改变的影响。方法雄性SD大鼠18只,体重180～220g,随机分成对照组和EGF组,选用大鼠坐骨神经切断近端双重结扎的动物实验模型。对照组于神经结扎近端注入生理盐水5μl,EGF组注入hEGF5μl(10μgEGF溶于5μl生理盐水中)分别于手术后7、14、28d用4%多聚甲醛灌注后取腰5背根神经节观察感觉神经元超微结构的改变。结果腰5背根神经节超微结构均发生改变,随着时间的推移,对照组内感觉神经元细胞核逐渐缩小,核质淡,稀疏,线粒体肿胀,嵴消失,基质丢失,卫星细胞从与神经元紧密相连到与之分离。EGF组神经元仅出现线粒体轻度的变性改变,其余结构基本正常。结论EGF对神经损伤后感觉神经元有一定的保护作用     

神经病理性痛大鼠背根神经节P2X3受体表达及电生理学特性的改变

目的 观察坐骨神经缩窄性损伤(CCI)致神经病理性痛后大鼠背根节神经元P2X3受体的表达和电生理学特性的变化.方法 应用免疫组化技术观察CCI后不同时间相应节段(L4、L5、L6)脊髓及背根神经节P2X3受体表达的变化;全细胞膜片钳技术观察CCI后不同时间L4、L5、L6节段背根节神经元ATP反应电流的变化.结果 CCI后7、14 d,损伤同侧L4、L5、L6节段背根节P2X3免疫阳性神经元数量明显增加,而且染色加深,相应节段的脊髓背角浅层P2X3免疫反应性亦明显增加.CCI后7、14 d,损伤同侧相应节段背根节产生ATP快反应电流的细胞数量明显增加,电流幅度亦增强.应用非选择性P2X受体拮抗剂Suramin能明显抑制ATP反应电流.结论 坐骨神经缩窄性损伤致神经痛后同侧对应节段的背根节神经元P2X3受体表达明显增加,ATP快反应电流明显增强.     

侧脑室注射改构体酸性成纤维细胞生长因子对帕金森病大鼠旋转行为的影响

目的 观察改构体酸性成纤维细胞生长因子(aFGF)对帕金森病大鼠模型行为学的影响.方法 采用SD雄性大鼠72只,随机分为假手术组,PD模型组,PD模型加生理盐水组(PD+NS组),PD模型加改构体aFGF组(PD+aFGF组),每组18只.通过脑内立体定向术,将6-OHDA注入后3组大鼠左侧黑质致密部(substantia nigra pars compacta,SNC)和中脑被盖腹侧区(ventral tegmeatal area,VTA)以建立帕金森病模型,PD+Ns组于术后第1天起每隔2d于右侧侧脑室注射10μl Ns,PD+aFGF组于术后第1天起每隔2d于右侧侧脑室注射10μl改构体aFGF(0.2μg/μl),假手术组只进行假手术处理,分别于术后4d,1,2,3,4周用阿扑吗啡(apomorphine,APO)给大鼠腹腔注射(0.5mg/kg),观察并记录大鼠行为学变化情况.结果 假手术组均未出现旋转行为,PD模型组和PD+Ns组大鼠旋转启动时间逐渐缩短,持续时间逐渐延长,旋转速度逐渐加快,至术后2周旋转行为趋于稳定;在术后2周,PD+aFGF组旋转启动时间[(5.50±1.18)min],较PD模型组[(3.60±1.17)min,P<0.05]和PD+Ns组[(3.10±1.02)min,P<0.05]明显延长;PD+aFGF组旋转持续时间[(25.90±8.80)min],较PD模型组[(55.40±10.14)min,P<0.05]和PD+Ns组[(53.90±12.27)min,P<0.05]明显缩短;PD+aFGF组旋转平均速度[(6.52±1.34)r/min],较PD模型组[(12.90±2.21)r/min,P<0.05]和PD+Ns组[(11.80±3.65)r/min,P<0.05]明显减慢.术后3周、4周,大鼠旋转行为各项指标统计学分析结果 与术后2周相同.结论 PD模型大鼠具有明显得行为学改变,改构体aFGF能明显改善大鼠的旋转行为.     

瑞舒伐他汀对急性冠状动脉综合征患者介入后血管内皮功能、血清炎症因子和预后的影响

目的：研究瑞舒伐他汀（RSVT）对急性冠状动脉综合征（ACS）患者行经皮冠状动脉介入治疗术（PCI）后血管内皮功能（VEF）、血清炎症因子（IF）以及预后的影响。方法选择该院2010年7月至2013年7月80例行PCI术的ACS病患进行观察,分为观察组和对照组,每组40例。观察组给予RSVT干预治疗,对照组给予常规治疗。对比两组VEF、IF以及预后情况。结果观察组术后4周VWF为（92．6±12．3）％、ET‐1为（55．6±5．6）ng／L;较对照组术后4周的（105．4±13．6）％、（67．8±7．4）ng／L明显更低;观察组术后4周NO为（78．6±9．4）μmol／L,较对照组术后4周的（63．2±9．5）μmol／L明显更高。观察组PCI术后4周CRP为（5．4±2．2）mg／L,较对照组的（10．5±4．1）mg／L明显更低。观察组CVE发生率为5．00％（2／40）、再狭窄率为2．50％（1／40）明显较对照组的30．00％（12／40）、10．00％（4／40）更低,差异有统计学意义（均P＜0．05）。结论RSVT能够有效改善ACS病患PCI术后VEF,减少炎性反应,降低CVE以及再狭窄的发生概率。