迷走神经电刺激对脑卒中运动功能障碍作用的研究进展

脑卒中是导致长期获得性残疾的主要原因。将康复与增强神经突触可塑性的技术相结合的干预措施有望增强脑卒中患者康复。近年来,迷走神经电刺激（vagus nerve stimulation, VNS）作为一种神经调节疗法在治疗脑卒中后运动功能障碍方面逐渐显示出积极作用。本文介绍了现有的VNS应用于脑卒中患者运动功能障碍康复的临床试验。     

RNA干扰c-kit对K562细胞成瘤性的影响

目的：研究短干扰RNA（siRNA）对K562细胞中c—kit基因表达的影响。方法：构建针对c—kit基因的shRNA表达质粒pSilencer—kit,在脂质体介导下转染人白血病细胞系K562,应用RT—PCR检测c—kitmRNA水平变化,并将转染后的K562细胞注射裸小鼠体内检测成瘤能力的变化。结果：重组体pSilencer-kit转染细胞后显著下调c—kitmRNA水平及其在裸鼠体内成瘤能力,与对照组相比差异有统计学意义（P〈0．01）.结论：利用RNA干扰技术能有效抑制靶基因c—kit的表达,为肿瘤的基因治疗提供新思路。     

α-7烟碱型乙酰胆碱受体激动剂EVP-6124的Ⅱb期临床试验获得阳性结果

EnVivo制药公司近日在美国神经精神药理学会第50届年会上公布了其研发的新型口服α-7烟碱型乙酰胆碱（Ach）受体激动剂EVP-6124的Ⅱb期临床试验阳性结果。     

花椒素对疼痛的改善作用及对炎症介质的影响

目的 探讨花椒素对炎性疼痛的改善作用及其调控机制。方法 构建昆明小鼠一般性疼痛模型和甲醛伤害性疼痛模型,比较花椒素对两种疼痛模型的影响;记录两种模型小鼠自主活动次数,并检测炎症因子水平;分析α7烟碱型乙酰胆碱受体(α7 nicotinic acetylcholine receptor,α7nAChR)拮抗剂对花椒素镇痛的阻断作用,并采用蛋白免疫印迹法测定α7nAChR拮抗剂阻断花椒素后JAK2及STAT3蛋白表达的变化。结果 一般性疼痛模型,注射花椒素1 h后,小鼠疼痛阈值由(17.2±4.8)s上升至(30.3±9.2)s,提高率由(43.3±1.3)%提高至(161.5±8.9)%;注射花椒素2 h后,小鼠疼痛阈值由(15.8±5.4)s提高至(30.6±7.5)s,提高率由(31.7±2.5)%提高至(158.7±9.8)%;甲醛伤害性疼痛模型,对照组与花椒素组Ⅰ相急性疼痛期出现舔足行为持续时间无明显差异,而Ⅱ相慢性疼痛期的小鼠疼痛行为持续时间缩短,且呈剂量依赖性;注射花椒素后,一般性疼痛模型小鼠自主活动次数由(198.2±71.8)次减少为(102.0±48.5)次,而甲醛伤害...     

α-芋螺毒素[A10L]PnIA荧光探针的设计合成及活性研究

α7烟碱型乙酰胆碱受体(nAChR)广泛分布于中枢和外周神经系统,与多种神经系统疾病、炎症反应密切相关.α-芋螺毒素[A 10L]PnIA作为靶向α7 nAChR的拮抗剂,对研究α7 nAChR相关生理、病理过程具有重要作用.利用荧光素5-羧基四甲基罗丹明标记[A 10L]PnIA,体外两步法氧化折叠获得活性肽([A1...     

白三烯拮抗剂ONO-1078对电刺激迷走神经、辣椒素和P物质引起的豚鼠心血管反应的作用(英文)

阐明ONO-1078(ONO,4-氧-8-[对-(4-苯丁氧基)苯甲酰氨基]-2-(5-四唑基)-4H-1-苯并毗喃)对神经原性刺激诱导心血管反应的作用.方法:观察豚鼠心房和心室伊文思蓝渗出以及平均主动脉压(MAP)变化.结果:在阿托品(1 mg·kg~(-1),iv)预先处理后,电刺激迷走神经(ESV,10 Hz,5 ms,2或10 V,90 s)显著增高伊文思蓝渗出;辣椒素和P物质也增加染料渗出并降低平均动脉压(MAP).ONO(0.03,0.1 mg·kg~(-1),iv)抑制ESV的反应,在刺激强度低(2 V)时更明显;ONO 0.03 mg·kg~(-1)减弱辣椒素引起的微血管渗漏和低血压,但对P物质无影响.结论:ONO-1078可能通过抑制感觉神经肽释放而调节神经原性炎症时的心血管反应.     

激活小胶质细胞α_7-nAChR减少Aβ蛋白所致的神经元毒性的研究

目的研究激活小胶质细胞上的α7-烟碱型乙酰胆碱受体对减少由于β-淀粉样蛋白1-42沉积所致的慢性炎性反应对神经元毒性作用。方法取孕18 d的胎鼠大脑皮质做原代神经元的培养,选用微管相关蛋白-2与神经元特异性烯醇化酶作为神经元特异性标志物,用免疫细胞化学技术对神经元进行鉴定。培养10 d的神经元与小胶质细胞共同培养于带有膜插件的培养皿中。试验分空白对照组、β-淀粉样蛋白组、共培养组、烟碱预处理组,各组添加β-淀粉样蛋白1-42共培养96 h后,用流式细胞仪及免疫荧光法检测神经元的凋亡率。结果体外培养10 d的神经元,用免疫荧光及DAB显色鉴定神经元均能显色,细胞纯度达到了94.49%。激活组与未激活组相比神经元的凋亡率明显下降(P<0.05)。结论激活小胶质细胞上的α7-烟碱型乙酰胆碱受体能减少β-淀粉样蛋白介导的神经元凋亡,对神经元产生保护作用。     

胆碱能抗炎通路α7烟碱型乙酰胆碱受体的最新研究进展

α7nAChR(α7 nicotinic acetylcholine receptor)属于烟碱型乙酰胆碱受体（nAChRs）,是机体胆碱能抗炎通路的关键受体,在免疫系统的神经调控中发挥重要作用。近期研究发现,α7nAChR还参与调节免疫以外的多种生理病理过程,如非酒精性脂肪肝、血管新生、保护心脏等。此外,α7nAChR在神经退行性疾病等与能量代谢紧密关联。本文综述α7nAChR在治疗炎症、改善能量代谢中发挥的作用,以及α7nAChR参与治疗的新方向。     

α7n型乙酰胆碱能受体在小胶质细胞中下调炎症水平的作用及其机制研究

目的 小胶质细胞在中枢神经系统的炎症相关疾病中发挥重要作用,旨在研究在炎症环境中α7 n型乙酰胆碱能受体的抗炎作用及机制.方法 应用PNU282987激动α7 n型乙酰胆碱能受体,应用脂多糖(LPS)造成细胞的炎症模型,通过实时定量PCR技术检测BV2细胞的炎症因子IL-1β、IL-6、TNF-α及M1型巨噬细胞标记物...     

屈洛昔芬对妊娠大鼠黄体细胞凋亡和C-myc、Bax、Bcl-2蛋白表达的影响(英文)

目的:观察屈洛昔芬对妊娠大鼠黄体细胞凋亡的影响,并分析黄体中C-myc,Bax和Bcl-2蛋白表达与屈洛昔芬所诱导的黄体细胞凋亡间的可能联系。方法:大鼠于妊娠第2天口服给予屈洛昔芬20mg·kg~(-1)后,分别于第4天和第8天取卵巢,HE染色和TUNEL法检测黄体中凋亡细胞的存在,免疫组织化学方法观察黄体中C-myc,Bax和Bcl-2蛋白的表达,同时测定卵巢重量、蛋白质含量及血中孕酮水平。结果:大鼠经屈洛昔芬处理后,第4天卵巢黄体中出现明显的凋亡细胞,第8天时更加明显。卵巢重量、蛋白质含量及血清孕酮水平在第8天时显著下降。屈洛昔芬处理组大鼠卵巢黄体中C-myc蛋白表达在第4天即显著增加,Bax蛋白表达的显著增加在第8天可观察到,而Bcl-2蛋白在卵巢黄体中的表达无明显改变。结论:屈洛昔芬可诱导妊娠大鼠着床前黄体细胞凋亡,C-myc蛋白及Bax/Bcl-2蛋白表达的增加可能与该过程有关。     

急慢性迷走神经刺激对大鼠海洛因复吸行为的影响及作用机制

目的:探讨急慢性迷走神经刺激(VNS)对大鼠海洛因复吸行为可能的干预作用及机制。方法:SD大鼠每天进行4h的海洛因自身给药训练,持续14d,建立具有强迫性觅药和给药特征的自身给药模型。随后所有大鼠植入VNS电极,恢复后进行10d的海洛因觅药行为消退训练,期间分组给予急性VNS、慢性VNS或假刺激。消退结束后测定各组大鼠条件性线索诱导下的海洛因觅药行为的恢复(即海洛因复吸行为)。免疫荧光法检测中枢核团c-Fos的表达水平。结果:海洛因复吸行为测定结果显示,与假刺激对照组比较,急性VNS组和慢性VNS组的有效鼻触数均明显降低(P<0.01)。免疫荧光的结果显示,与对照组比较,急性VNS组(P<0.05)和慢性VNS组(P<0.01)中央杏仁核(Ce)的c-Fos表达水平均明显降低,而边缘下区(IL)的c-Fos表达水平均明显升高(P<0.05)。结论:急慢性VNS均能够显著抑制大鼠的海洛因复吸行为,其机制可能与Ce和IL脑区神经元激活的改变有关。     

The role of the α7 subunit of the nicotinic acetylcholine receptor on motor coordination in mice treated with methyllycaconitine and anabasine

The adverse effects of methyllycaconitine (MLA) have been attributed to competitive antagonism of nicotinic acetylcholine receptors (nAChR). Research has indicated a correlation between the LD₅₀ of MLA and the amount of α₇ nAChR in various mouse strains, suggesting that mice with more α₇ nAChR require more MLA to be poisoned. However, recent research demonstrated that there was no difference in the acute lethality (LD₅₀) to MLA in mice lacking the α₇ nAChR subunit compared with wild‐type mice. The objective of this study was to determine if the α₇ nAChR subunit plays a role in motor coordination deficiencies that result from exposure to nAChR antagonists and agonists. We compared the motor function and coordination in wild‐type mice to mice lacking the α₇ subunit of the nAChR, after treating them with a non‐lethal dose of MLA or anabasine, using the following tests: balance beam, grip strength, rotarod, open field and tremor monitor. Analysis of the data indicated that overall there was no difference between the wild‐type and knockout mice (P = 0.39 for grip strength; P = 0.21 for rotarod; P = 0.41 for balance beam; P = 0.22 for open field; and P = 0.62 for tremors). Thus results from this study suggest that α₇ nAChR does not play an integral role in the acute effects of MLA or anabasine on motor function/coordination. Consequently other subunits of nAChRs found in the neuromuscular junction are likely the primary target for MLA and anabasine resulting in motor coordination deficiencies and acute toxicosis. Published 2013. This article is a U.S. Government work and is in the public domain in the USA.     

1,2,5,6-4H-1-烷基-3-取代吡啶类新衍生物对血管内皮细胞功能的调节作用及其构效关系

目的：以槟榔碱为结构母核，设计合成1，2，5，6-4H-1-烷基-3-取代吡啶类新衍生物，分析其对血管内皮细胞功能的影响。方法：采用离体大鼠主动脉环和离体豚鼠回肠收缩实验，分别观察新结构化合物对血管和回肠平滑肌张力的影响；并进一步观察一氧化氮合酶抑制剂L-NAME、环氧酶抑制剂吲哚美辛、M受体亚型非选择性激动剂毛果芸香碱和M受体亚型非选择性拮抗剂阿托品对新结构化合物诱发的血管舒张反应的影响。结果：从100个新化合物中发现4个有舒血管反应，但不激活M受体的化合物，分别是HH91、HH95、HH98、HH103。新化合物HH103诱发的内皮依赖性舒血管反应可被L-NAME所拮抗，但不被吲哚美辛、阿托品和毛果芸香碱拮抗。结论：新化合物可诱发内皮依赖性舒张反应并具有特定构效关系；HH103通过促进内皮细胞释放NO发挥其效应；但其作用特征又与乙酰胆碱不同。     

Effect of caffeine on response of rabbit isolated corpus cavernosum to high K+ solution, noradrenaline and transmural electrical stimulation

1. Caffeine has wide-ranging activities on smooth muscles, including contractile and relaxant effects. The aim of the present study was to examine the activity of caffeine on rabbit corpus cavernosum (RCC). 2. The effects of caffeine (0.5-4.0 mmol/L) on the response of RCC to high K+ solution, noradrenaline (NA) and transmural electrical stimulation (EFS) were studied in a tissue bath system. 3. Caffeine did not contract the RCC. However, 0.5-4.0 mmol/L caffeine caused concentration-dependent relaxation of tension development in high-K+ (120 mmol/L) solution in contrast with the solvent control. At 4.0 mmol/L caffeine, high-K+ solution-induced tone of the RCC was reduced by 73.4 +/- 7.3%. Caffeine (0.5-4.0 mmol/L) also concentration-dependently relaxed NA (12.5 micro mol/L)-induced tonic contraction of the RCC. At 4.0 mmol/L caffeine, NA-induced tone of the RCC was reduced by 41.1 +/- 7.0%. Incubation of RCC in 2.0 mmol/L caffeine for 30 min prior to EFS (1-40 Hz) caused a marked rightward shift in the frequency-response curve. 4. The results of the present study suggest that caffeine exhibits relaxant activity on rabbit cavernosal smooth muscle and the mechanism of this activity possibly involves inhibition of Ca2+ signalling.     

龙胆苦苷对大鼠血管平滑肌细胞增殖、迁移的影响

目的 观察龙胆苦苷对血管平滑肌细胞（vascular smooth muscle cells,VSMC）增殖、迁移能力的影响,并探讨其可能的作用机制。方法 体外分离培养大鼠血管平滑肌细胞,将血管平滑肌细胞随机分为分为control组、GPS处理组、PF-3758309+GPS组。采用噻唑蓝(MTT)法观察各组细胞增殖能力,计算细胞增殖抑制率;采用细胞划痕实验观察各组细胞迁移能力,计算细胞迁移率。采用Western blotting法检测各组细胞Pak4及MMP-2表达情况。结果 与对照组相比,龙胆苦苷处理组细胞增殖率及迁移率明显降低（P<0.01）,GPS组细胞内Pak4、MMP-2 表达水平明显降低（P<0.01）;而与GPS组相比,PF-3758309+GPS组可明显增加细胞增殖率及迁移率（P<0.05）,血管平滑肌细胞内 MMP-2 表达水平明显上调（P<0.05）。 结论 龙胆苦苷具有抑制血管平滑肌细胞增殖及迁移,其机制可能与抑制Pak4相关。     

Effects of electrical stimulation of the reticular formation and chlorpromazine on performance of trace conditioned avoidance response in the rat

The effects of electrical stimulation to the mesencephalic reticular formation and chlorpromazine on the performance of a trace conditioned avoidance response by rats were studied. Either treatment alone impaired the performance; this impairment was a function of level of stimulation or dose of the drug, respectively. The performance deficit was not present when a high intensity of stimulation of the reticular formation was combined with a moderate dose of chlorpromazine. However, the combination of a high dose of the drug with a low stimulation intensity interfered with the avoidance responding more than any other condition tested. These effects appeared to be independent of neutral or negative reinforcement effects of the stimulation, as tested in an independent situation.     

Effects of clonidine,dihydralazine and splanchnic nerve stimulation on the release of neuropeptide Y,MET-enkephalin and catecholamines from dog adrenal medulla

Summary Various neuropeptides are costored together with catecholamines in the adrenal medulla. The concurrent release (evaluated by adrenal vein plasma levels) of these neuropeptides [neuropeptide Y (NPY), met-enkephaline (ME)] and catecholamines [adrenaline (A) and noradrenaline (NA)] from the adrenal gland was examined in chloralose-anesthetized dogs after intravenous administration of clonidine (10 g/kg) and dihydralazine (1 mg/kg). These results were compared to those obtained after the stimulation of the right splanchnic nerve at 1, 5 and 10 Hz frequencies. The increment in the release of catecholamines and neuropeptides was evaluated for dihydralazine and splanchnic nerve stimulation.Dihydralazine (at its maximal effect) induced a significant preferential increase in catecholamines (expressed as mean (SEM): NA: 17.3 (5.4) fold, A: 13.1 (2.6) fold) and ME (16.0 (7.1) fold) versus basal values. However, the significant increase in NPY-LI was only 2.0 (0.4) times the baseline.Splanchnic nerve stimulation induced a frequency-dependent increase in catecholamines and neuropeptides. When the stimulation frequency was increased from 1 Hz to 5 Hz, NA and A levels increased 17.9 (4.3) and 14.0 (2.2) fold, respectively and ME levels 14.1 (3.0) fold. By contrast, NPY-LI was increased only 2.3 (0.3) fold under the same conditions. Increasing the stimulation frequency from 5 Hz to 10 Hz resulted in similar elevations of NA, ME, and NPY-LI adrenal plasma levels (about 4 times) whereas A only increased twice.Clonidine decreased catecholamine and ME adrenal plasma levels (the maximal percent decrease when compared with control values was about 75%) whereas NPY adrenal plasma levels remained unchanged.In conclusion, the present data indicate that (i) both adrenal ME and NA always exhibit corelease in a parallel fashion which is not the case for NPY art; (ii) different populations of chromaffin vesicles could be preferentially mobilized according to different physiological and pharmacological patterns. Correspondence to M. A. Tran at the above address     

Effects of hexamethonium and other ganglionic blocking agents on electrical activity of the esophagus induced by vagal stimulation in the dog.

Electrical activity of the dog's esophagus was evoked by electrical stimulation of the vagus nerve and recorded by using a suction electrode placed on the serosal surface of the esophagus. Neuromuscular blocking agents (d-tubocurarine and succinylcholine) blocked this activity, while atropine had no effect. Hexamethonium (in doses which did not effect neuromuscular transmission in the diaphragm) and high doses of nicotine significantly depressed the evoked esophageal electrical activity, while lower doses of nicotine facilitated the response. The results suggest that a certain proportion of the striated muscle of the esophagus is innervated by processes from the intermediate ganglion cells in the Auerbach plexus.     

TF3-H4对CD4~+CD25~+调节性T细胞和CD4~+CD25~-效应性T细胞早期活化的影响

目的观察1',2',3',7'-四氢茶黄素-3,3'-双没食子酸酯(TF3-H4)对CD4+CD25+调节性T细胞和CD4+CD25-效应性T细胞早期活化的影响,分析TF3-H4对两群功能相反的CD4+T细胞的作用。方法免疫磁珠分离BALB/c小鼠脾脏CD4+CD25+调节性T细胞和CD4+CD25-效应性T细胞,加入ConA或PDB,和TF3-H4共同孵育12 h后收集细胞,流式细胞仪分析细胞表面早期活化标志CD69的表达。结果在ConA和PDB的作用下,两群细胞早期活化标志CD69的表达均升高。TF3-H4(20 mg.L-1)不能抑制PKC激动剂PDB激活的CD4+CD25-效应性T细胞CD69的表达,却能抑制ConA激活的CD4+CD25-效应性T细胞CD69表达;同时,TF3-H4也能明显抑制ConA激活的CD4+CD25+调节性T细胞的CD69表达。结论茶黄素衍生物TF3-H4可能经由TCR活化途径的上游抑制CD4+CD25-效应性T细胞活化;TF3-H4对CD4+CD25+调节性T细胞和CD4+CD25-效应性T细胞早期活化的抑制作用,可能是该类化合物发挥抗炎、抗肿瘤作用的机制之一。