速激肽NK-1受体拮抗剂SR-140333对抗原引起致敏大鼠气道高反应性的影响

为观察速激肽NK-1受体拮抗剂SR-140333对抗原攻击引起的致敏大鼠气道高反应性的影响,测定了致敏大鼠在抗原攻击前后的基础呼吸频率,对MCh的反应性及支气管-肺泡灌洗液中的白细胞数量。实验结果显示,致敏大鼠吸入OA后6h基础呼吸频率增加,并显著增加乙酰甲胆碱(MCh)的反应性、MCh的-logPC₃₀值和支气管-肺泡灌洗液中的白细胞数量。ip速激肽NK-1受体拮抗剂SR-140333(0.1mg·kg^-1)或地塞米松(0.5mg·kg^-1),可明显抑制上述反应,小剂量SR-140333(0.01mg·kg^-1)仅有部分抑制作用。结果提示抗原攻击可引起致敏大鼠气道高反应性和气道炎症,速激肽NK-1受体拮抗剂可抑制这些反应。     

速激肽NK-1受体拮抗剂SR-140333对抗原引起致敏大鼠气道高反应性的影响

为观察速激肽ＮＫ１受体拮抗剂ＳＲ１４０３３３对抗原攻击引起的致敏大鼠气道高反应性的影响，测定了致敏大鼠在抗原攻击前后的基础呼吸频率，对ＭＣｈ的反应性及支气管肺泡灌洗液中的白细胞数量。实验结果显示，致敏大鼠吸入ＯＡ后６ｈ基础呼吸频率增加，并显著增加乙酰甲胆碱（ＭＣｈ）的反应性、ＭＣｈ的－ｌｏｇＰＣ３０值和支气管肺泡灌洗液中的白细胞数量。ｉｐ速激肽ＮＫ１受体拮抗剂ＳＲ１４０３３３（０１ｍｇ·ｋｇ－１）或地塞米松（０５ｍｇ·ｋｇ－１），可明显抑制上述反应，小剂量ＳＲ１４０３３３（００１ｍｇ·ｋｇ－１）仅有部分抑制作用。结果提示抗原攻击可引起致敏大鼠气道高反应性和气道炎症，速激肽ＮＫ１受体拮抗剂可抑制这些反应     

高乌甲素对手术致痛大鼠P物质的影响

新型镇痛药高乌甲素已越来越多地用于临床术后镇痛。以往研究高乌甲素多是在化学性致痛模型中进行,本文采用大鼠切口疼痛模型,观察高乌甲素对手术致痛大鼠P物质的影响,探讨高乌甲素镇痛的量效关系及给药时机。     

喷他佐辛对慢性缩窄性损伤模型大鼠皮肤P物质及神经激肽1受体表达的影响

目的观察喷他佐辛对坐骨神经慢性缩窄性损伤(CCI)模型大鼠疼痛阈值、皮肤中P物质(SP)及神经激肽1受体(NK1-R)表达的影响。方法 36只雄性SD大鼠随机分为对照组、模型组和治疗组,每组12只。暴露模型组和治疗组大鼠坐骨神经给予三线点扎,观测得造模后第15日达疼痛高峰期。治疗组于造模后第15日单次给予喷他佐辛10 mg·kg-1股部肌内注射,其他2组不给药。观察各组大鼠疼痛阈值变化,采用免疫组化法和RT-PCR法检测大鼠皮肤中SP及NK1-R蛋白和mRNA的表达水平。结果与造模前比较,造模后第15日模型组和治疗组疼痛阈值均显著降低(P <0.05),且均低于对照组(P <0.05)。治疗组大鼠治疗后疼痛阈值显著升高(P <0.05),且高于模型组(P <0.05)。模型组SP及NK1-R蛋白和mRNA的表达水平显著高于对照组(P <0.05),治疗组SP及NK1-R蛋白和mRNA的表达水平显著低于模型组(P<0.05)。结论喷他佐辛对CCI模型大鼠坐骨神经慢性疼痛有明显的镇痛效果,其机制可能与降低皮肤中SP及NK1-R的表达有关。     

阿瑞匹坦对恶性肿瘤神经激肽-1受体和P物质表达的影响北大核心CSCD

目的研究P物质及神经激肽-1受体(NK-1R)在非小细胞肺癌(NSCLC)组织及人肺腺癌细胞系A549细胞系中的表达特点,探讨NK-1R拮抗药阿瑞匹坦对NK-1 R、P物质表达的影响。方法收集临床NSCLC患者原发瘤组织,相应癌旁组织和相对正常肺组织标本,用免疫组织化学法检测P物质和NK-1R的表达情况,用免疫细胞化学染色检测二者在A549细胞系的表达特点。用MTT比色法检测不同浓度阿瑞匹坦作用后A549细胞生长增殖的变化情况。结果非小细胞肺癌组织中P物质、NK-1R表达阳性率明显高于癌旁组织和正常肺组织(P<0.05)。NSCLC组织中,P物质表达阳性部位主要位于肺肿瘤细胞胞浆,而NK-1R在肿瘤细胞胞浆和胞膜均有表达。肺癌A549细胞系中绝大多数细胞均有P物质、NK-1R表达。加入阿瑞匹坦培养的A549细胞P物质、NK-1R表达明显下降(P<0.05)。阿瑞匹坦在浓度为1×10-6mol·L-1时对A549细胞生长增殖的抑制作用最明显。结论 P物质和NK-1R在NSCLC组织和A549细胞系中高表达,提示神经内分泌机制参与了NSCLC的发生和发展过程。NK-1受体拮抗药可抑制P物质和NK-1R的阳性表达,且能抑制A549细胞生长增殖,NK-1受体可成为治疗NSCLC的新靶点。     

不同给药方式对P2X_1受体介导大鼠肠系膜动脉收缩反应的影响

目的研究不同方式给予α,β-MeATP对肠系膜动脉P2X1受体介导血管收缩反应的影响。方法制备大鼠离体肠系膜动脉环标本,采用非累积给药法和单浓度给药法两种方式给予α,β-MeATP,记录药物诱发的等长收缩反应。结果两种给药方式给予α,β-MeATP(10-7-10-4mol·L-1)均可使大鼠离体肠系膜动脉产生浓度依赖性收缩反应。以KCl最大收缩反应或以标本湿重标化α,β-MeATP诱发的收缩反应时,α,β-MeATP单浓度给药的收缩反应均大于非累积给药(P<0.01)。以标本湿重标化时,10-4mol·L-1浓度α,β-MeATP诱发的收缩反应分别是(0.73±0.10)g·mg-1(单浓度组)和(0.38±0.05)g·mg-1(非累积组);以KCl最大收缩反应标化时,分别是(53.17±6.0)%(单浓度组)和(36.78±5.71)%(非累积组)。在α,β-MeATP非累积给药组120mmol·L-1KCl诱发的动脉收缩反应小于单浓度给药组和NA对照组(P<0.01)。结论在大鼠肠系膜动脉,非累积给予α,β-MeATP降低P2X1受体介导的收缩反应以及高钾诱发的收缩反应,该给药方式可能导致错误的实验结论 。     

氟比洛芬酯对手术致痛大鼠行为学的影响

目的观察氟比洛芬酯对手术致痛大鼠行为学的影响。 方法将48只SD大鼠随机分为6组。对照1（D1）组大鼠单纯吸入异氟醚麻醉,不进行手术,其余5组（D2、K1～K4组）以异氟醚吸入使大鼠麻醉,按BRENNAN 法手术,建立大鼠手术切口疼痛模型。20 min后经尾静脉给药。D1组、D2组大鼠给予0.9%氯化钠注射液,K1～K4组分别给予氟比洛芬酯注射液2,4,8,16 mg·kg 1。待大鼠手术清醒及每组给药后1 h,进行累积疼痛评分,观察给药后反应。结果氟比洛芬酯可剂量依赖性地降低切口疼痛模型大鼠的累积疼痛评分(P＜0.05)。 结论氟比洛芬酯可抑制外科手术的疼痛反应,并呈一定的剂量依赖性。     

利多卡因对致痛大鼠L4～5脊髓P物质分布影响的实验研究

目的研究鞘内(IT)利多卡因对福尔马林致痛大鼠脊髓P物质(SP)含量的影响,探讨其镇痛的可能机制.方法取雄性SD大鼠15只,分为三组假手术组、对照组和治疗组.后足皮下注射3%福尔马林50ml致痛造膜.对照组造膜前IT0.9%氯化钠20μl;治疗组造模前IT2%利多卡因20μl.采用免疫组织化学SABC法结合图像分析技术.结果大鼠脊髓背角Ⅰ层和Ⅱ层有密集SP分布,治疗组与对照组相比含量明显减少,差异非常显著(P＜0.01).结论在大鼠外周炎性疼痛模型中,超前鞘内给利多卡因的镇痛作用与其抑制SP在脊髓背角的释放有关.     

在线SPE-HPLC技术检测血浆中NK-1浓度的研究

目的：建立NK-1的在线SPE-HPLC血浆样品检测方法。方法：采用在线SPE-HPLC技术测定小鼠血浆中NK-1浓度。预处理柱：CAPCELL PAK MF Ph-1色谱柱（4.6mm×10mm）,色谱分析柱：Venusil MP C18（5μm,100,4.6mm×150mm）,预处理流动相：20mM磷酸盐缓冲液（pH值=2.8）,分析流动相：20mM磷酸盐缓冲液（pH值=2.8）∶乙腈（70∶30,v/v）,检测波长：270nm,柱温：30℃。结果：本方法线性范围25～1000ng·ml-1,最低检测限2ng·ml-1,r=0.9994;日内、日间精密度分别小于5.0%和6.0%;高、中、低3个浓度的QC样品：方法回收率分别为103.25±2.98%、97.21±4.06%、102.35±2.64%。结论：本法测定血浆中NK-1含量准确、简便,适用于NK-1药物动力学的研究。     

NK-1 receptor antagonists as antitumor drugs: a survey of the literature from 2000 to 2011

Introduction: After binding to the neurokinin (NK-1) receptor, substance P (SP) induces tumor cell proliferation, the migration of tumor cells (invasion and metastasis) and angiogenesis. By contrast, NK-1 receptor antagonists inhibit tumor cell proliferation (tumor cells die by apoptosis), block the migratory activity of tumor cells and exert antiangiogenic properties.

Areas covered: This review offers a 12-year overview of the underlying mechanism of the action of the SP/NK-1 receptor system and NK-1 receptor antagonists in cancer, providing a new approach to the treatment of tumors.

Expert opinion: Chemically diverse NK-1 receptor antagonists have been identified. The antitumor action of these compounds is independent of their chemical structures and such action is associated with their affinity for the NK-1 receptor and with the dose of the antagonist administered. The NK-1 receptor can be considered as a target in cancer treatment and NK-1 receptor antagonists could be considered as new antitumor drugs. The NK-1 receptor antagonist aprepitant is used in clinical practice and exerts an antitumor action against tumor cells in vitro. In the future, such antitumor action should be tested in human clinical trials.     

Smooth muscle of rabbit isolated aorta contains the NK-2 tachykinin receptor

Summary Neurokinin A, neurokinin B and substance P caused concentration-related contractions of rabbit isolated aorta with pD₂ values of 8.1, 6.9 and 6.0, respectively. [D-Pro², D-Trp^7,9]-substance P, a competitive tachykinin antagonist, had pA₂ values of 5.3 against neurokinin A, 5.1 against neurokinin B and 5.2 against substance P indicating that tachykinin receptors mediated responses to the agonists. [pGIu⁵,MePhe⁸,-McGly⁹]-substance P 5–11 (DiMe-C7), senktide and septide did not contract the aorta. It is concluded that of the known tachykinin receptors smooth muscle of the rabbit isolated aorta contains only the NK-2 type. Send offprint requests to J. W. Constantine at the above address     

NK-1 as a melanoma target

Introduction: Melanoma expresses both neurokinin-1 (NK-1) receptors and substance P (SP). After binding to the NK-1 receptor, SP induces tumor cell proliferation in melanoma cells, whereas after binding to the same NK-1 receptor, antagonists inhibit melanoma cell proliferation and cause tumor cell death by apoptosis. Thus, the NK-1 receptor could be a new and promising target in melanoma therapy.

Areas covered: This review provides an overview of the underlying mechanism of action of the SP/NK-1 receptor system, and NK-1 receptor antagonists in human melanoma, over the last 7 years.

Expert opinion: In stages III – IV, no effective treatment exists for melanoma and hence there is an urgent need to improve therapy in melanoma patients. The NK-1 receptor is a promising new target in human melanoma treatment, since preclinical assays (most of them in vitro assays) have reported that NK-1 receptor antagonists exert an antitumor action against melanoma. The NK-1 receptor antagonist aprepitant is used in clinical practice and exerts an antitumor action against human melanoma in vitro. In the future, such antitumor action should be tested in human clinical trials. This should be faster compared with less investigated NK-1 receptor antagonists, because a great part of the required safety and characterization studies for aprepitant have already been carried out.     

Synthesis and purification of two SP (6–11) analogues with enhanced selectivity for the NK-1 receptor

Two hexapeptide analogues of Substance P (6–11) have been synthesized. Replacement of Gly₉ by proline provides a peptide with tenfold enhanced selectivity for the NK-1 receptor. The corresponding proline-containing glycopeptide incorporating a β-d -glucopyranosyl residue linked to the side-chain of Glus was 100 times more selective than Substance P for the same receptor.     

The inflammatory reaction induced by formalin in the rat paw

The involvement of bradykinin and some other inflammatory mediators in formalin-induced oedema and plasma extravasation was examined. Formalin was injected in rat paws at two doses, 1.75% or 5%. The lower dose induced the development of an immediate oedema associated with a progressive accumulation of ¹²⁵I-labelled albumin in the paws. These changes were suppressed by pretreatment with capsaicin or xylocaine. They were abolished by RP67580, a NK₁ receptor antagonist, and increased by phosphoramidon or diprotin A. They were not affected by HOE140, a bradykinin B₂ antagonist, captopril, methysergide, mepyramine, indomethacin, ketoprofen or l-N ^G-nitroarginine. The higher dose of formalin induced a swelling of the paws which took place in two phases associated with two periods of increase in vascular permeability. This oedema was reduced by pretreatment with capsaicin but not with xylocaine. It was reduced by RP67580 injected before or 30 min after formalin. It was inhibited by mepyramine, methysergide, indomethacin and NS-398, a cyclooxy-genase-2 inhibitor. It was not modified by HOE140. Its development was similar in normal and kininogen-deficient rats. We concluded that formalin administered at a low dose induces an oedema which mainly results from a neurogenic inflammation mediated by neuropeptides such as substance P. At higher doses, formalin induces an oedema which mainly depends on the release of substance P, prostanoids, 5-hydroxytryptamine and histamine. Bradykinin plays no significant role in the vascular changes whereas this peptide has been reported to participate in the stimulation of nociceptive afferent neurons. This discrepancy could be explained by a difference in the threshold of stimulation of the nociceptive neurons and that of the cells of the vascular walls, or by a formation of kinins in close contact of the neurons. Received: 29 June 1998 / Accepted: 20 November 1998     

Analgesic effects of mGlu1 and mGlu5 receptor antagonists in the rat formalin test

mGlu1 and mGlu5 receptors have been implicated in pain associated with inflammation. In the present study, the formalin test was used to measure sustained pain with components of tissue injury. The aims of the present study were to assess: (i) the role of mGlu1 and mGlu5 receptors in inflammatory pain using selective antagonist EMQMCM, 1.25-5 mg/kg, as the mGlu1 receptor antagonist, and MPEP or MTEP, 2.5-10 mg/kg, as mGlu5 receptor antagonist; (ii) the possible interaction between mGlu1 and mGlu5 receptor antagonists and morphine; and (iii) whether tolerance develops to the analgesic effects of these antagonists after prolonged treatment. EMQMCM, MTEP and MPEP significantly reduced the manifestation of both phases of formalin response. However, all these mGlu receptor antagonists did not affect the withdrawal latencies in a model of acute pain (Hargreaves test), which has a different underlying mechanism. In the present study, the suppressive effect on formalin-induced pain behaviour was much stronger when mGlu1 and mGlu5 receptor antagonists were co-injected compared to administration of a single antagonist, but this effect was not seen when mGlu receptor antagonist was co-administered with morphine. This is in contrast to the pronounced inhibitory effects after co-treatment with morphine and the uncompetitive NMDA receptor antagonist memantine. The present study also provides the first direct in vivo evidence that prolonged administration of MTEP (5 mg/kg) over 7 days leads to the development of tolerance to its antinociceptive effects. Such tolerance was not observed when EMQMCM (5 mg/kg) was administered in the same manner. In conclusion, these results provide additional arguments for the role of group I mGlu receptors in pain with inflammatory conditions.     

NKP608, an NK1 receptor antagonist, has an anxiolytic action in the social interaction test in rats

RATIONALE: Evidence is starting to accumulate that NK1 receptor antagonists might have anxiolytic effects in animal tests and in patients. OBJECTIVE: To examine the effects of NKP608, a substance P antagonist acting at NK1 receptors, in various conditions of the social interaction test of anxiety and to determine its effects after 3 and 6 weeks of treatment. METHODS: Rats were tested after vehicle, 0.01 or 0.1 mg/kg PO in three conditions of the social interaction test that varied in the level of anxiety generated. Thus pairs of rats were tested in an arena with which they were unfamiliar that was lit by high (HU) or low (LU) light and in the condition that generated the lowest level of anxiety, i.e. an arena with which they were familiar, lit by low light (LF). They were also tested after 3 and 6 weeks of treatment with 0.03 mg/kg and after 24 h withdrawal from these chronic treatments. RESULTS: NKP608 had significant anxiolytic effects at 0.01, 0.03 and 0.1 mg/kg PO in the HU and LU test conditions, but was without effect in the LF condition, except for an increased incidence of bite attacks at 0.1 mg/kg. The anxiolytic effect of 0.03 mg/kg remained after 3 weeks of chronic treatment and there was no anxiogenic effect after 24 h of drug withdrawal. Following 6 weeks of chronic treatment (0.03 mg/kg per day), tolerance had developed, but no anxiogenic withdrawal effect was seen 24 h after the last dose. CONCLUSIONS: These results provide further evidence that substance P may play a role in mediating states of anxiety and suggest that the selective NK1 receptor antagonist NKP608 may prove a useful anxiolytic compound.     

Yohimbine produces antinociception in the formalin test in rats: involvement of serotonin1A receptors

Rationale: Previous studies have suggested that the α₂-adrenergic receptor antagonist yohimbine produced antinociceptive effects in the formalin test in rats. However, yohimbine is also an agonist at serotonin (5-HT)_1A receptors, suggesting the possibility that the antinociceptive effects of yohimbine might be mediated via these receptors. Objective: The purpose of the present studies was to evaluate the potential role of 5-HT_1A receptors in mediating the antinociceptive effects of yohimbine. Methods: The antinociceptive effects of yohimbine were evaluated using the formalin test in rats. Results: Yohimbine (2.5–10 mg/kg s.c.) produced dose-related antinociception during both phase I and phase II of the formalin test, and was approximately equipotent and equiefficacious to morphine. The selective 5-HT_1A receptor antagonist WAY 100,635 (0.03–3.0 mg/kg s.c.) produced a partial reversal of yohimbine. In comparison, the selective 5-HT_1A receptor agonist (±)8-hydroxy- dipropylaminotetralin HBr (8OH-DPAT; 1.0 mg/kg s.c.) also produced a dose-related antinociception in the formalin test, although 8OH-DPAT was completely reversed by WAY 100,635 (3.0 mg/kg s.c.). The antinociceptive effects of yohimbine were not antagonized by the 5-HT_1B/1D antagonist GR 127935 (1.0 mg/kg and 3.0 mg/kg s.c.), the 5-HT₂ antagonist LY53857 (1.0 mg/kg s.c.), or the 5-HT₃ antagonist zatosetron (3.0 mg/kg s.c.). Conclusions: The present results demonstrate that yohimbine produces a dose-related antinociception in the formalin test in rats which is mediated in part by the agonistic actions at 5-HT_1A receptors. Received: 10 September 1999 / Final version: 5 November 1999     

Synthesis and biological activity of Substance P C-terminal hexapeptide and heptapeptide analogues

The analogues [Glu(OBzl)¹¹]SP_6–11 and [Glu(OBzl)¹¹]SP_5–11 of the C-terminal hexapeptide and heptapeptide of Substance P have been synthesized by conventional solution methods. In each analogue the SCH₃ group of Met¹¹ is replaced by the COOCH₂C₆H₅ group. The in vitro activity of both analogues has been determined on three biological preparations: guinea pig ileum (GPI), rat vas deferens (RVD), and rat portal vein (RPV). The selectivity for the different receptors has been studied by utilizing atropine-treated guinea pig ileum (GPI + At). The results showed that both analogues are mainly active on GPI through the NK-1 receptor and that both analogues are equipotent to Substance P.     

NK-3 receptors mediate enhancement of substance P release from capsaicin-sensitive spinal cord afferent terminals

1. The effects of NK-3 receptor agonists on the release of substance P-immunoreactivity (SP-LI) have been investigated using superfused rat spinal cord synaptosomes.
2. The Ca²⁺-dependent overflow of SP-LI evoked by 35 mM KCl was concentration-dependently enhanced by senktide (EC₅₀=52 nM; maximal effect=70%) or [MePhe⁷]NKB (EC₅₀=5.5 nM; maximal effect=125%), both selective agonists at receptors of the NK-3 type.
3. The potentiation of the SP-LI overflow elicited by 100 nM senktide or [MePhe⁷]NKB was prevented by the NK-3 receptor antagonist (+)-SR142801. The antagonist halved, at 10 nM, and almost abolished, at 100 nM, the effect of both agonists. The effect of senktide or [MePhe⁷]NKB was insensitive to antagonists at NK-1 or NK-2 receptors.
4. Capsaicin (0.1–1 μM) stimulated SP-LI release in a concentration-dependent manner from spinal cord synaptosomes. The SP-LI overflow elicited by 1 μM capsaicin was completely dependent on external Ca²⁺. Senktide could not affect the capsaicin-evoked release of SP-LI.
5. Senktide failed to potentiate the K⁺-evoked overflow of SP-LI from synaptosomes previously exposed for 15 min in superfusion to capsaicin.
6. The results show that release-enhancing NK-3 receptors are located on axon terminals of capsaicin-sensitive primary afferent neurones in the spinal cord. Antagonists at NK-3 receptors might help controlling pain transmission.