拟胆碱药物对金环蛇、银环蛇毒中毒小鼠的解毒活性研究

目的测定拟胆碱药物对金环蛇、银环蛇毒中毒小鼠是否具有解毒活性。方法于昆明小鼠腹腔注射800μg/kg金环蛇毒(1.8×LD_(50))或50μg/kg银环蛇毒(1.4×LD_(50)),2 h后于腹腔注射生理盐水或不同剂量拟胆碱药物,观察24 h内对照组及拟胆碱药物组的小鼠死亡时间及死亡率,统计分析拟胆碱药物组是否具有解毒活性。结果注射金环蛇毒后,生理盐水组小鼠的死亡时间为[(3.64±1.03)～(5.08±1.27)]h,死亡率为70.0%～100%。溴吡斯的明组(62.5、125、250μg/kg),甲硫酸-新斯的明组(62.5、125、250μg/kg),毒扁豆碱组(62.5、125、250μg/kg)及石杉碱甲组(62.5、125、250μg/kg)的小鼠死亡时间分别为[(4.45±0.94)～(6.48±3.27]h、[(2.79±0.74)～(4.17±1.44)]h、[(3.74±2.21)～(5.62±2.47)]h及[(4.43±2.55)～(4.62±1.49]h;醋甲胆碱组(5、20、80μg/kg),氯贝胆碱组(5、20、80μg/kg),他克林组(250、500、1000、2000μg/kg),加兰他敏组(125、250、500、1000、2000μg/kg)及多奈哌齐组(500、1000、2000μg/kg)的小鼠死亡时间分别为[(4.59±1.90)～(5.30±2.18)]h、[(3.25±0.75)～(4.53±1.72)]h、[(5.09±2.57)～(5.66±2.63)]h、[(3.48±0.80)～(5.30±2.51)]h及[(3.25±1.47)～(4.67±1.51)]h,各给药组的死亡时间及死亡率(80%～100%)与对照组比较无明显差异(P0.05)。与对照组相比,银环蛇毒各给药组小鼠的死亡时间及死亡率亦无明显差异性(P0.05)。结论拟胆碱药物不能显著延长金环蛇、银环蛇毒中毒小鼠的死亡时间或提高生存率,无显著解毒活性。     

天麻素对脑缺血再灌注小鼠海马新生神经元的保护作用

目的:探讨不同剂量天麻素(Gastrodin,GAS)对缺血再灌注模型小鼠海马新生神经元的保护作用及其可能机制。方法:将50只C57BL/6小鼠随机分为假手术组(Sham)、模型组(MCAO+Vehicle)、模型+天麻素低剂量(10 mg/kg)组(MCAO+GAS(L)),模型+天麻素中剂量(50 mg/kg)组(MCAO+GAS(M)),模型+天麻素高剂量(100 mg/kg)组(MCAO+GAS(H))。除Sham组接受假手术外(皮肤切开,分离颈动脉),分别对MCAO组及MCAO+GAS各组行右侧颈内动脉栓线术,造成并保持大脑中动脉闭塞(MCAO)1 h。术后,Sham组和MCAO组即刻腹腔注射生理盐水(0.1 m L/kg),MCAO+GAS各组注射不同剂量天麻素,每24小时重复给药1次,连续7天。最后一次注射24 h后,对各组小鼠的神经功能进行评分,之后处死动物取脑组织,通过HE、免疫荧光染色以及Western Blot观察和比较各组小鼠海马形态学和DCX的表达水平。结果:(1)术后第7天,MCAO+Vehicle组小鼠神经功能评分(3.2±0.63)显著高于假手术组、MCAO+GAS(M)(1.8±0.63)和MCAO+GAS(H)组(P0.05)。(2)术后第7天,与假手术组相比较,MCAO+vehicle组海马颗粒细胞排列不规则,核稍大且固缩深染,齿状回部有较多空洞;与MCAO+vehicle组比较,MCAO+GAS(H)组海马空洞样改变减少,细胞排列较整齐,胞膜较完整,核结构较清晰。(3)术后第7天,与假手术组相比较,DCX染色阳性细胞数显著减少,而MCAO+GAS(M)组DCX染色阳性细胞数(183±64.5)和MCAO+GAS(H)组DCX染色阳性细胞数(195±93.68)显著高于MCAO+Vehicle组。MCAO+Vehicle组海马的DCX表达水平显著低于假手术组及MCAO+GAS(M)组和GAS(H)组(P0.01)。结论:天麻素可能通过上调海马DCX的表达水平,调节海马神经发生,进而对缺血性脑卒中后再灌注发挥神经保护作用。     

角叉菜胶诱导的慢性血栓小鼠模型的建立

目的采用不同浓度的角叉菜胶溶液连续腹腔注射的方式处理小鼠,筛选慢性血栓小鼠模型的条件。方法将70只ICR雄性小鼠随机分为7组,每组10只,即:空白组对照组、0.02%剂量组、0.04%剂量组、0.06%剂量组、0.08%剂量组、0.10%剂量组、0.20%剂量组。空白组小鼠腹腔注射生理盐水(0.01 mL/g),其余组腹腔注射相应浓度的角叉菜胶溶液(0.01 mL/g),每天注射1次;各组小鼠饲养于(20±1)℃室温下并自由采食、饮水。每天观察各组出现黑尾的情况(黑尾率、黑尾长度);出栓即采样;采血检测血凝四项,检测结肠组织与血浆中氧化应激指标及炎性因子指标。结果 0.06%剂量组小鼠第5天出栓2只,第6天出栓8只;与空白对照组相比,0.06%剂量组小鼠FIB含量显著升高(P 0.05),APTT显著缩短(P 0.05),TT显著延长(P 0.05),PT极显著延长(P 0.01); 0.06%剂量组结肠组织与血浆中SOD、GSH-Px均极显著降低(P 0.01)、MDA显著升高(P0.05); 0.06%剂量组结肠组织与血浆中TNF-α、IL-1β显著升高(P0.05)、IL-10显著降低(P 0.05)。结论初步得出室温为(20±1)℃条件下,0.06%的角叉菜胶溶液(0.01 mL/g)连续腹腔注射6 d(每天1次)是建立慢性血栓小鼠模型的最适条件。     

脓毒症诱导建立小鼠凝血指标与炎症细胞因子的变化及临床意义

目的:探讨脓毒症诱导建立的免疫缺陷型脓毒症小鼠的凝血指标与炎症细胞因子的变化及临床意义。方法:以40只SPF级雄性SCID小鼠(BALB/c背景)为研究对象,按实验要求随机分为对照组和观察组,观察组建立脓毒症小鼠模型(腹腔注射脂多糖1.5 mg/kg),对照组腹腔注射等体积的生理盐水,8小时后收集小鼠血清并分离肝组织,检测凝血指标(血清凝血酶原时间、活化的部分凝血酶时间)与血清炎症细胞因子(tumor necrosis factor-α,TNF-α;Interleukin-10,IL-10;interferon-γ,INF-γ)水平,并进行肝组织病理学评分,并分析观察组小鼠的凝血指标、炎症细胞因子与肺组织病理评分的相关性。结果:腹腔注射后8小时,观察组小鼠的肝组织病理评分为6.44±0.87分,对照组小鼠为0.46±0.09分,显著低于观察组,两组对比差异有统计学意义(P0.05)。观察组小鼠的血清TNF-α、IL-10、INF-γ水平都明显高于对照组小鼠(P0.05),其APTT、TT值分别为27.56±4.45s和18.69±1.62s,都明显少于对照组小鼠(30.45±2.14s和19.99±1.55s),差异都有统计学意义(P0.05)。直线相关分析显示观察组小鼠的肺组织病理评分与血清TNF-α、IL-10、INF-γ水平呈现明显正相关性(P0.05),与APTT、TT呈现明显负相关性(P0.05)。结论:脓毒症诱导建立的免疫缺陷型小鼠固有免疫细胞活性增强,可出现失控的炎症反应与凝血功能异常,使得小鼠肺组织损伤加重。     

兔脓毒性休克模型的建立

目的:建立相对稳定的兔脓毒性休克模型.方法:雄性新西兰大白兔20只,随机分为模型组和假手术组.麻醉后无菌操作下取中、下腹部正中切口,找出盲肠,距盲肠末端8.0cm处4号丝线结扎盲肠和相应血管;于盲肠游离末端避开血管戳孔2次,孔径0.5cm;将盲肠原位放回腹腔,缝合腹壁切口;腹腔内注射30ml/kg温生理盐水.假手术组只探查腹腔.术后每隔3小时监测肛温.颈动脉插管有创动脉压监测确定模型成功后送血培养、血气、血乳酸,监测4小时后处死动物,送腹水培养,取右肺中叶测定肺含水量,取心、肝、脾、肺、肾、肠常规切片HE染色.结果:制模成功时间为:(18.91±1.384)小时;模型平均动脉压MAP(95.00±10.817 vs 52.38±15.565,P＜0.05);血气:PH(7.40±0.047 vs 7.09±0.146,P＜0.01),PCO2(30.0±5.831 vs 19.80±4.104,P＜0.01),BE(-6.46±4.931 vs -24.11±4.276,P＜0.01),HCO3-(18.45±4.367 vs 5.73±2.422,P＜0.01);血乳酸(2.53±1.108 vs 7.85±5.834,P＜0.05);血培养(7/10)阳性:大肠埃希菌;腹水培养(10/10)阳性:大肠埃希菌及阴沟肠杆菌;肛温于术后呈先上升后下降,假手术组肛温较模型组差异有显著性(39.63±0.492 vs 36.82±0.999,P＜0.01);成模后肛温与平均动脉压呈正相关关系,r=0.748.P=0.013,方程:y(平均动脉压)=34.46x+0.45;肺干湿比及肺含水量无明显差异[(0.22±0.014 vs 0.19±0.288,P＞0.05),(78.17±1.375 vs 80.58±2.878,P＞0.05)].常规HE染色可见明显病理学改变.结论:本模型可模拟多微生物感染致脓毒性休克模型,模型相对稳定,可用于兔种属.     

不同注射途径的脂肪间充质干细胞对大鼠肠炎疗效的实验研究

目的探讨脂肪间充质干细胞(ADMSCs)腹腔及静脉注射对三硝基苯磺酸(TNBS)诱导肠炎疗效的影响。方法将32只大鼠完全随机分成正常组、腹腔注射组、静脉注射组及模型组,每组8只。用TNBS诱导炎症性肠病(IBD)动物模型,腹腔及静脉注入ADMSCs,记录大鼠的疾病活动指数(DAI)、观察结肠宏观损伤及微观变化、测定结肠的髓过氧化物酶(MPO)活性、检测结肠Ki-67^+细胞的表达、比较血液中IL-1β、TNF-α浓度及结肠TGF-β、IL-6、IL-17A、IL-10的基因表达水平。采用单因素方差分析及独立t检验进行统计学分析。结果 ADMSCs腹腔注射组(98.05±0.63)g高于静脉注射组[(94.32±0.48)g,t=12.281,P=0.000],同时腹腔注射组DAI评分1.71±0.75低于静脉注射组3.57±0.97,(t=-3.980,P=0.002)。另外发现腹腔注射组结肠组织内髓过氧化物酶MPO浓度(95.75±5.52)U/g低于静脉注射组(74.37±5.12)U/g,(t=-7.513,P=0.000),腹腔注射组结肠病理评分2.14±0.69低于静脉注射组3.57±0.76,(t=-3.612,P=0.004)。结肠免疫荧光检查发现腹腔注射组比静脉注射组有更多的Ki-67^+细胞。酶联免疫吸附测定(ELISA)发现腹腔注射组血浆中IL-1β的浓度(130.71±7.08)pg/ml比静脉注射组(163±9.09)pg/ml低,(t=-8.518,P=0.000),同样腹腔注射组血浆中TNF-α的浓度(201.71±6.75)pg/ml也比静脉注射组(242.28±8.30)pg/ml低,(t=-10.033,P=0.000)。此外,结肠组织实时定量聚合酶联反应(RT-qPCR)的结果显示腹腔注射组IL-6 mRNA的表达4.34±0.48比静脉注射低6.15±1.05,(t=-4.147,P=0.001),腹腔注射组IL-17A mRNA的表达2.61±0.53也比静脉注射低3.57±0.46,(t=-4.301,P=0.001)。然而,腹腔注射组IL-10 mRNA的表达水平37.75±4.46比静脉注射组高27.68±2.25,(t=5.327,P=0.001),腹腔注射组TGF-β mRNA的表达水平15.82±0.99也比静脉注射组高11.97±2.25,(t=3.740,P=0.003)。结论 ADMSCs腹腔注射优于静脉注射并可能成为ADMSCs治疗IBD的较好选择。     

脂多糖联合MPTP诱导的小鼠慢性帕金森病模型的评价

目的建立脂多糖(lipopolysaccharide,LPS)联合1-甲基-4-苯基-1,2,3,6-四氢吡啶(1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine,MPTP)诱导的慢性小鼠帕金森病模型,探讨其行为学和黒质多巴胺能神经元细胞的变化情况。方法将20只C57BL/6小鼠随机分为模型组和对照组,模型组每日腹腔注射LPS(0.25 mg/kg)一次,连续3 d,最后一次注射LPS 4 h后,每日腹腔注射MPTP(25 mg/kg)一次,连续2 d,对照组腹腔注射相同量的生理盐水。8周后用步态分析和转棒实验评价小鼠的行为学能力,免疫组化法观察黑质酪氨酸羟化酶(TH)阳性细胞变化情况。结果模型组小鼠行为学变化较对照组差异有显著性(P0.05),模型组小鼠中脑黒质区显示出严重的神经元细胞损伤。结论 LPS联合MPTP腹腔注射可成功诱导出慢性小鼠帕金森病模型,提示该模型可用于帕金森病的发病机制及药物治疗效果等相关研究。     

儿童功能性便秘患者肠道菌群及血清脑肠肽水平的变化

目的探讨儿童功能性便秘(FC)患者肠道菌群及血清脑肠肽水平的变化。方法选取2017年1月至2019年4月在我院儿科门诊治疗的80例FC患儿为研究组,选择同期于体检中心检查的无胃肠道疾病的30例健康儿童为健康组。研究组患儿根据病情轻重分为轻中度组(52例)和重度组(28例)。比较各组儿童肠道菌群数量[乳杆菌、双歧杆菌和大肠埃希菌]及血清脑肠肽指标[一氧化氮(NO)、P物质(SP)和血管活性肠肽(VIP)]水平的差异。结果研究组患儿肠道双歧杆菌[(8.16±0.89)lg CFU/g]和乳杆菌数量[(7.92±0.83)lg CFU/g]明显少于健康组[(8.94±0.97)lg CFU/g、(8.63±0.91)lg CFU/g],而大肠埃希菌数量[(8.12±0.81)lg CFU/g]明显多于健康组[(7.30±0.72)lg CFU/g],差异有统计学意义(t=2.37、2.35、2.44,P0.05)。重度组患儿肠道双歧杆菌和乳杆菌数量明显少于轻中度组,而大肠埃希菌数量明显多于轻中度组(t=2.36、2.34、2.29,P0.05)。研究组患儿血清NO和VIP水平[(50.12±8.25)μmol/L、(29.17±5.12)pg/mL]明显高于健康组[(34.46±6.13)μmol/L、(20.43±4.79)pg/mL],SP水平[(5.21±0.82)ng/mL]明显低于健康组[(7.57±1.04)ng/mL],差异有统计学意义(t=2.78、2.54、2.47,P0.05)。重度组患儿血清NO和VIP水平高于轻中度组,SP水平明显低于轻中度组(t=2.31、2.41、2.23,P0.05)。结论 FC患儿存在肠道菌群失调和脑肠肽分泌异常,且随着病情的加重其肠道菌群失调及脑肠肽分泌异常越发明显。     

不同时间睡眠干扰所致小鼠的类抑郁样行为

目的研究不同时间睡眠干扰后所致的小鼠类抑郁样行为学表现。方法使用滚筒睡眠干扰仪对小鼠进行不同时间(5、10、15 d)睡眠干扰后,分别对各组动物进行自主活动测试实验、强迫游泳实验和悬尾实验。结果干扰5 d组与对照组比,自主活动总路程、平均速度、运动总时间减少(P<0.05),干扰10 d组与对照组比,自主活动总路程、平均速度、运动总时间减少(P<0.05)。干扰5 d后,小鼠强迫游泳、悬尾实验中各指标与对照组比无差异,干扰10 d组小鼠的游泳不动时间[(143.92±9.48)s]和悬尾不动时间[(127.89±6.33)s]均较对照组小鼠游泳不动时间[(128.50±6.63)s]和悬尾不动时间[(102.64±9.57)s]长(P<0.05),干扰15 d组小鼠的游泳不动时间[(143.08±8.13)s]和悬尾不动时间[(119.10±10.43)s]均较对照组小鼠游泳不动时间[(113.00±7.28)s]和悬尾不动时间[(89.55±9.07)s]长(P<0.05)。干扰5 d组、10 d组、15 d组与对照组比,体重均降低(P<0.05)。结论滚筒法睡眠干扰10、15 d可引起小鼠的类抑郁样行为学表现。     

灵芝多糖对顺铂引起的呕吐具抑制作用

以雄性昆明小鼠为实验材料,研究灵芝多糖对顺铂引起呕吐的抑制作用。实验分为5组,生理盐水+生理盐水组:腹腔注射0.9% NaCl两次,格拉司琼+顺铂组:腹腔注射格拉司琼和顺铂,生理盐水+顺铂组:腹腔注射0.9% NaCl和顺铂,灵芝多糖+生理盐水组:腹腔注射灵芝多糖和0.9% NaCl,灵芝多糖+顺铂组:腹腔注射灵芝多糖和顺铂,两种成分的注射间隔为30min。上述处理每天1次,连续5d,比较各组小鼠对高岭土的摄取量和脑内Fos蛋白表达水平。结果显示:顺铂可增加小鼠高岭土摄取量和脑内Fos蛋白表达水平,而格拉司琼和灵芝多糖可减少小鼠对高岭土的摄取量并降低脑内Fos蛋白表达水平。因此认为灵芝多糖可有效抑制顺铂引起的恶心呕吐。     

SR142801 behaves as a tachykinin NK-3 receptor agonist on a spinal nociceptive reflex in the rat

Effects of two commonly used tachykinin NK-3 receptor antagonists (SR 142801 and R820) intrathecally (i.t.) administered were assessed in the rat tail-flick test. SR142801 and its (R)-enantiomer SR142806 (1.3, 6.5 and 65 nmol) were found as potent as senktide and [MePhe7]NKB (NK-3 selective agonists) to induce transient antinociceptive effects. Naloxone (10 microg) and R820 (6.5 nmol) blocked reversibly the responses to 6.5 nmol senktide, [MePhe7]NKB, SR142801 and SR142806 when administered i.t. 15 min earlier. However, the antinociceptive responses induced by SR142801 and SR142806 were not affected by i.t. pretreatments with NK-1 (6.5 nmol SR140333) and NK-2 (6.5 nmol SR48968) receptor antagonists. In control experiments, the NK-1 and NK-2 antagonists prevented the hyperalgesic effects to NK-1 ([Sar9,Met(O2)11]SP) and NK-2 ([beta-Ala8] NKA(4-10)) receptor agonists (6.5 nmol i.t.), respectively. R820 had no direct effect on nociceptive threshold and failed to alter angiotensin II-induced antinociception. The data suggest that the antinociceptive effect of SR142801 is due to an agonist effect at NK-3 receptor in the rat spinal cord that involves a local opioid mechanism. These results can be best explained by the existence of inter-species NK-3 receptor subtypes.     

Receptor subtypes involved in tachykinin-mediated edema formation.

Intradermal (ID) injection of the natural tachykinins substance P (SP), neurokinin A (NKA), and neurokinin B (NKB) (0.3-30 nmol) resulted in a marked and dose-related rat paw edema, with mean ED50 values of 2.68 nmol, 1.17 nmol, and 0.80 nmol, respectively. The ID injection of the selective NK1, SP methyl-ester (1-30 nmol), NK2, [beta-Ala8]-neurokinin A4-10) (beta-Ala, 0.3-30 nmol), or NK3, senktide (1-10 nmol) agonists, caused extensive edema formation with mean ED50s of 0.48 nmol, 0.41 nmol, and 0.18 nmol, respectively. The ID injection of the selective NK1 antagonist FK888 (0.1-3 nmol) produced marked inhibition (94%, 52%, and 66%, respectively) of rat paw edema induced by SP, NKA, or SP methyl-ester. The ID co-injection of the NK2 receptor antagonist SR 48968 elicited a graded inhibition (52%, 67%, and 35%, respectively) of rat paw edema induced by NKA, beta-Ala and, to a lesser extent, the edema caused by SP. Finally, the ID co-injection of the NK, receptor antagonist SR 142801 significantly inhibited (53%, 76%, 53%, and 100%, respectively) the edema formation caused by NKB and NKA or by SP and senktide. Together, the data of the present study suggest that tachykinin-mediated rat paw edema depends on the activation of NK1, NK2 and NK3 receptor subtypes, with apparent major involvement of NK1 receptors subtypes.     

Neurokinin A and neurokinin B in the human retina

Very recently, the authors found levels of neurokinin (NK) A-like immunoreactivities in the human retina which were more than five times higher than those of substance P (SP). The present study aimed to find out how many of these immunoreactivities can be attributed to NKA and NKB and then the exact distribution pattern of both NKA and NKB was evaluated in the human retina and compared with that of SP. For this purpose, NKA-like immunoreactivities were characterized in the human retina by reversed phase HPLC followed by radioimmunoassay using the K12 antibody which recognizes both NKA and NKB. Furthermore, the retinae from both a 22- and 70-year-old donor were processed for double-immunofluorescence NKA/SP and NKB/SP. The results showed that NKA contributes to approximately two thirds and NKB to approximately one third of the immunoreactivities measured with the K12 antibody. NKA was found to be localized in sparse amacrine cells in the proximal inner nuclear layer, in displaced amacrine cells in the ganglion cell layer with processes ramifying in stratum 3 of the inner plexiform layer and also in sparse ganglion cells. By contrast, staining for NKB was only observed in ganglion cells and in the nerve fiber layer. Double-immunofluorescence revealed cellular colocalization of NKA with SP and also of NKB with SP. Thus, the levels of NKA and NKB are more than three and two times higher than those of SP, respectively. Whereas the distribution pattern of NKA is typical for neuropeptides, the localization of NKB exclusively in ganglion cells is atypical and unique.     

Pharmacological profile of hemokinin 1: a novel member of the tachykinin family

Recently, the cloning of a novel preprotachykinin gene (PPT-C) has been reported. This gene codes for a novel peptide named hemokinin 1 (HK-1). In contrast with the known tachykinins, which are exclusively expressed in neuronal tissues, PPT-C mRNA was detected primarily in hematopoietic cells. In this study, we pharmacologically characterised the effects of HK-1 using three tachykinin monoreceptor systems, namely the rabbit jugular vein (rbJV) for NK(1), the rabbit pulmonary artery (rbPA) for NK(2), and rat portal vein (rPV) for NK(3) receptors. In all these preparations substance P (SP), neurokinin A (NKA) and neurokinin B (NKB) elicited concentration dependent contractions showing similar maximal effects and the following rank order of potency: SP > NKA = NKB in the rbJV, NKA > NKB > SP in the rbPA, and NKB > NKA > SP in the rPV. In those vessels HK-1 behaved as a full agonist displaying potencies similar (rbPA and rPV) or slightly higher (rbJV) than those of SP. In the rbJV, SR 140333, a selective NK(1) receptor antagonist, antagonised the effects of HK-1 and SP with similar high potencies (pK(B) 9.3 and 9.5, respectively). Similar results were obtained with the pseudopeptide NK(1) antagonist, MEN 11467 (pK(B) 8.8 and 8.6, respectively). Taken together, these data indicate that HK-1 behaves as a NK(1) preferring receptor agonist.     

[3H]Neurokinin B and 125I-Bolton Hunter Eledoisin Label Identical Tachykinin Binding Sites in the Rat Brain

[3H]Neurokinin B ([3H]NKB) of high specific activity (75 Ci/mmol) was synthesized for study of its binding to crude synaptosomes from the rat cerebral cortex. The specific binding of [3H]NKB (75% of total binding) was temperature dependent, saturable, and reversible. Scatchard analyses and Hill plots showed the existence of a single population of noninteracting binding sites (KD = 4.3 nM; Bmax = 123 fmol/mg of protein). Competition studies indicated the following rank order of potencies among tachykinins: NKB greater than eledoisin (E) greater than kassinin greater than physalaemin greater than neurokinin A (NKA) greater than substance P (SP), a result suggesting that NKB might be the endogenous ligand for [3H]NKB binding sites. It is of interest that 127I-Bolton Hunter (BH) NKA (127I-BHNKA) was much more potent than NKA in inhibiting the specific binding of [3H]NKB, which raises certain questions concerning the use of 125I-BHNKA as a ligand for NKA binding sites in the brain. These results, as well as those obtained with different SP analogues, show a close similarity to those obtained previously with 125I-BHE binding to cortical synaptosomes. This suggested that the two ligands labeled identical binding sites. In addition, using either [3H]NKB or 125I-BHE as ligands, similar displacement curves were obtained with increasing concentrations of NKB and 127I-BHE. The similarity of the [3H]NKB and 125I-BHE binding sites was further confirmed by comparison of their localization on rat brain sections by autoradiography. The distribution of binding sites for [3H]NKB and 125I-BHE was identical throughout the brain, and the highest density of binding sites for the two ligands was found in layers IV and V of the cerebral cortex, the paraventricular nucleus of the hypothalamus (magnocellular part), and the ventral tegmental area.     

Heterogeneity of tachykinin receptors in the rabbit lung

The tachykinins, substance P (SP) and neurokinin A (NKA), are agonists for the NK₁ and NK₂ receptors, respectively. Tachykinins have various respiratory effects, including bronchoconstriction. This study characterizes tachykinin binding sites in the rabbit lung. We hypothesize that (2-[¹²⁵I]iodohistidyl¹)Neurokinin A ([¹²⁵I]NKA) interacts with NK₁ and NK₂ binding sites in the rabbit lung. The K_d determined from saturation isotherms was 0.69 X/÷1.14 nM (geometic mean X/÷ SEM) and the B_max was 4.15±0.22 femtomole/mg protein (arithmetic mean±SEM). Competitive inhibition studies with NKA, SP and various selective tachykinin agonists showed the rank order of potency: [β-Ala⁸]-Neurokinin A 4–10=SP ≫ NKA ≫ [Sar⁹,Met(O₂)¹¹]-Substance P. [β-Ala⁸]-Neurokinin A 4–10, a selective NK₂ agonist, and SP inhibition of [¹²⁵I]NKA binding were best described using a two-site model. Competitive inhibition studies using the selective nonpeptide NK₂ antagonist (SR 48968) and the selective nonpeptide NK₁ antagonist (CP 96,345) revealed K_i's of 5.5 nM and 8.1 nM, respectively. Our data therefore suggest that [¹²⁵I]NKA binds to both the NK₁ and NK₂ receptors in the lung. Special issue dedicated to Dr. Kinya Kuriyama.     

Different subtypes of tachykinin NK(1) receptor binding sites are present in the rat brain

(2-[(125)I]iodohistidyl(1))Neurokinin A ([(125)I]NKA), which labels "septide-sensitive" but not classic NK(1) binding sites in peripheral tissues, was used to determine whether septide-sensitive binding sites are also present in the rat brain. Binding studies were performed in the presence of SR 48968 (NK(2) antagonist) and senktide (NK(3) agonist) because [(125)I]NKA also labels peripheral NK(2) binding sites and, as shown in this study, central NK(3) binding sites. [(125)I]NKA was found to label not only septide-sensitive binding sites but also a new subtype of NK(1) binding site distinct from classic NK(1) binding sites. Both subtypes of [(125)I]NKA binding sites were sensitive to tachykinin NK(1) antagonists and agonists but also to the endogenous tachykinins NKA, neuropeptide K (NPK), and neuropeptide gamma (NPgamma). However, compounds of the septide family such as substance P(6-11) [SP(6-11)] and propionyl-[Met(O(2))(11)]SP(7-11) and some NK(1) antagonists, GR 82334, RP 67580, and CP 96345, had a much lower affinity for the new NK(1)-sensitive sites than for the septide-sensitive sites. The hypothalamus and colliculi possess only this new subtype of NK(1) site, whereas both types of [(125)I]NKA binding sites were found in the amygdala and some other brain structures. These results not only explain the central effects of septide or SP(6-11), but also those of NKA, NPK, and NPgamma, which can be selectively blocked by NK(1) receptor antagonists.     

酮替芬联合沙丁胺醇气雾剂喷吸对咳嗽变异性哮喘患者嗜酸性粒细胞趋化因子与肺功能的影响

目的:探讨酮替芬联合沙丁胺醇气雾剂喷吸对咳嗽变异性哮喘患者嗜酸性粒细胞趋化因子与肺功能的影响。方法:选择2015年3月到2016年4月我院接诊的98例咳嗽变异性哮喘患者研究,按抽签法将其随机分为观察组和对照组,每组49例。对照组患者给予沙丁胺醇气雾剂、氨茶碱治疗,观察组在对照组基础上给予酮替芬治疗。观察比较治疗前后两组临床疗效;白天及夜间咳嗽症状评分;嗜酸性粒细胞(EOS)计数和嗜酸性粒细胞趋化因子(Eotaxin)水平;肺功能相关评价指标(FVC、FEV1、PEF)。结果:治疗后,观察组有效率(93.88%)高于对照组(79.59%),差异有统计学意义(P0.05)。两组白天及夜间咳嗽症状评分均显著下降,观察组下降更为明显[(0.77±0.39)VS(0.99±0.52)、(0.87±0.32)VS(1.07±0.34)],差异有统计学意义(P0.05);两组EOS计数和Eotaxin水平均明显下降,观察组下降更为显著[(188.47±30.39)VS (232.59±30.52)、(169.44±27.79)VS (191.07±34.34)],差异具有统计学意义(P0.05);两组相关评价指标(FVC、FEV1、PEF)均明显提高,观察组相关指标值提高更为显著[(3.99±0.39)VS(3.87±0.12)、(3.87±0.79)VS(3.52±0.39)、(7.99±1.98)VS(7.34±1.01)],差异有统计学意义(P0.05)。结论:酮替芬联合沙丁胺醇气雾剂喷吸对咳嗽变异性哮喘的疗效显著,能明显降低患者嗜酸性粒细胞趋化因子的水平,显著改善患者的肺功能,值得临床推广运用。     

SR 142801, a tachykinin NK(3) receptor antagonist,prevents beta(2)-adrenoceptor agonist-induced hyperresponsiveness to neurokinin A in guinea-pig isolated trachea

We investigated whether fenoterol was able to enhance contractile responsiveness to neurokinin A (NKA) on the guinea-pig isolated trachea. We then studied the effects of two inhibitors of nuclear factor kappa B (NFkappaB), gliotoxin and pyrrolidine dithiocarbamate, and of the tachykinin NK(1), NK(2) and NK(3) receptor antagonists, SR 140333, SR 48968 and SR 142801 and determined whether tachykinin receptor gene expression was up-regulated in the trachea after exposure to fenoterol. Fenoterol (0.1 microM, 15 h, 21 degrees C) induced an increased contractile response to NKA (mean of difference in maximal tension between control and fenoterol +/- S.E.M; +0.47 +/- 0.14 g, n = 26, P < 0.01). This hyperresponsiveness was strongly reduced by co-incubation with gliotoxin (0.1 microg/ml) or pyrrolidine dithiocarbamate (0.1 mM) and abolished by SR 140333 (0.1 microM) and SR 142801 (0.1 microM). SR 48968 (0.1 microM) diminished the tracheal contractility to NKA but failed to reduce the hyperreactivity induced by fenoterol. Tachykinin NK(1) receptor (NK(1)R), NK(2) receptor (NK(2)R) and NK(3) receptor (NK(3)R) gene expression was analyzed by semiquantitative RT-PCR. Compared to control tissues, NK(1)R and NK(2)R mRNA expression was increased by about 1.6-fold and 1.4-fold, respectively, in tissues treated with fenoterol. We were unable to detect the presence of NK(3)R mRNA in the guinea-pig trachea. In conclusion, fenoterol induces tracheal hyperresponsiveness to NKA and an up-regulation of NK(1)R and NK(2)R gene expression. The hyperresponsiveness implicates the NFkappaB pathway and is abolished by tachykinin NK(1) (SR 140333) and NK(3) (SR 142801) receptor antagonists.     

Effects of Qufeng Xuanfei Decoction in Animal Model of Post-Infectious Cough

This study evaluated the effects and potential mechanisms of Qufeng Xuanfei decoction in animal model of post-infectious cough. Sixty SD rats were randomly divided into six groups (10 animals per group): control, disease model, low- (4.62 g kg^?1), medium- (9.24 g kg^?1), and high-dose (13.86 g kg^?1) decoction, and positive treatment groups (dextromethorphan hydrobromide, 8 mL kg^?1). To model post-infectious cough, all but control group animals were challenged with exposure to 50 g sawdust and 10 cigarette smokes for 30 min day^?1 for a total of 10 days, followed by subsequent exposures to lipopolysaccharide (20 µg) and capsaicin (10^?4 M) aerosols. The drugs were given by oral gavage for 15 days after which lung pathology, cell counts and cell differentials in bronchoalveolar lavage (BAL), and concentrations of neuropeptides [substance P (SP), neurokinins A (NKA) and B (NKB), and calcitonin gene-related peptide (CGRP)] in BAL (ELISA) were assessed. Compared with control group animals, significant inflammation and damage to bronchial epithelium were observed in the disease model group. A marked decrease in BAL percentages of all types of inflammatory cells was observed in the decoction-treated groups, with most changes in the medium-dose decoction group (p < 0.001 vs. disease model group). Further, airway inflammation and damage, as well as the levels of SP, NKA, NKB, and CGRP in BAL decreased the most in the medium-dose group (p < 0.001 vs. disease model group). In conclusion, medium-dose Qufeng Xuanfei decoction efficiently decreases the levels of neuropeptides, attenuates airway inflammation, and promotes recovery from disease.