P物质对变应性鼻炎动物模型鼻粘膜上皮短路电流的影响

目的 观察Ｐ物质（ＳＰ）作用于变应性鼻炎动物模型鼻粘膜引起的离子分泌改变导致的上皮表面短路电流的变化。方法 以鸡蛋清蛋白致敏大鼠后，用Ｕｓｓｉｎｇ室技术测定鼻粘膜上皮表面短路电流。并观察神经激肽受体（ＮＫ１）拮抗剂ＣＰ９６３４５、组胺Ｈ１受体拮抗剂吡拉明、Ｈ２受体拮抗剂雷尼替丁和河豚毒素对ＳＰ的阻断作用。结果 经ＳＰ刺激，致敏大鼠鼻粘膜表面Ｉｓｃ显著增高。经四种物质预处理均能显著阻断ＳＰ引起的Ｉｓ     

大鼠变应性鼻炎模型鼻粘膜P物质受体mRNA的表达

目的 探讨大鼠鼻粘膜Ｐ物质受体（ｓｕｂｓｔａｎｃｅ Ｐ ｒｅｃｅｐｔｏｒ,ＳＰ－Ｒ）ｍＲＮＡ的表达水平及其与变应性鼻炎的关系。方法 选健康Ｗｉｓｔａｒ大鼠２０只,雌雄不限,随机分为实验组和对照组各１０只。实验组用卵清蛋白行腹腔注射基础致敏,继之鼻局部激发建立大鼠变应性鼻炎模型,以β－肌动蛋白（β－ａｃｔｉｎ）作内标准,利用逆转录－多聚酶链反应（ｒｅｖｅｒｓｅ ｔｒａｎｓｃｒｉｐｔｉｖｅ ｐｏｌｙｍ     

辣椒辣素治疗实验性变应性鼻炎的组织病理学及免疫组织化学研究

目的　明确辣椒辣素（ｃａｐｓａｉｃｉｎ） 对实验性变应性鼻炎的治疗作用。方法　采用甲苯２ ,４二异氰酸酯（ｔｏｌｕｅｎｅ２ ,４ｄｉｉｓｏｃｙａｎａｔｅ,ＴＤＩ） 致敏新西兰兔制作变应性鼻炎动物模型, 观察给予辣椒辣素处理前后症状和体征的变化,同时应用组织病理学和免疫组织化学方法观察鼻粘膜的改变。结果应用辣椒辣素处理后, 症状和体征明显改善。免疫组织化学表明,辣椒辣素处理后Ｐ 物质免疫反应（ｓｕｂｓｔａｎｃｅ Ｐｉｍｍｕｎｏｒｅａｃｔｉｖｅ,ＳＰＩＲ） 纤维减少, 而对照组中明显增加。ＨＥ 染色显示辣椒辣素处理后,鼻粘膜未见水肿及炎性细胞浸润, 对照组均为鼻粘膜水肿、血管扩张及炎细胞浸润。结论　辣椒辣素能缓解流涕及喷嚏, 减少ＳＰＩＲ 纤维密度和数量, 而达到治疗变应性鼻炎的目的。     

辣椒辣素治疗实验性变应性鼻炎的组织病理学及免疫组织 …

目的 明确辣椒辣素对实验性变应性鼻炎的治疗作用。方法 采用甲苯－２,４－二异氰酸酯（ＴＤＩ）致敏新西兰兔制作变应性鼻炎动物模型,观察给予辣椒辣素处理前后症状和体征的变化,同时应用组织病理学和免疫组织化学方法观察鼻粘膜的改变。结果 应用辣椒辣素处理后,症状和体征明显改善。免疫组织化学表明,辣椒辣素处理后Ｐ物质免疫反应（ＳＰ－ＩＲ）纤维减少,而对照组中明显增加。ＨＥ染色显示辣椒辣素处理后,鼻粘膜未见水     

大鼠变应性鼻炎模型鼻粘膜三种肽能神经末梢免疫组化研究

为研究变应性鼻炎（ＡＲ）时鼻粘膜速激肽类神经纤维末梢的密度变化，用二异氰酸甲苯酯（ＴＤＩ）滴鼻建立ＷＳ系大鼠的ＡＲ实验模型，取鼻粘膜作免疫组化染色并测定三种速激肽类即Ｐ物质（ＳＰ）、神经激肽Ａ（ＮＫＡ）及神经激肽Ｂ（ＮＫＢ）神经纤维的密度。结果证实实验组动物鼻粘膜ＳＰ、ＮＫＡ及ＮＫＢ神经纤维的密度显著高于对照组（Ｐ＜０．０１）；实验组鼻粘膜三种肽能神经纤维染色加重、纤维变粗、曲张体增大。由于大鼠ＡＲ模型鼻粘膜ＳＰ、ＮＫＡ及ＮＫＢ神经纤维的密度明显增加，提示速激肽在ＡＲ的发病中起重要作用。     

分枝杆菌多糖促人LAK细胞增殖及对喉癌HEP－2细胞杀伤活性的研究

为探讨分枝杆菌多糖（ｍｙｃｏｂａｃｔｅｒｉａｌｐｏｌｙｓａｃｃｈａｒｉｄｅｓ，ＭＰＳ）在体外对ＬＡＫ细胞（ｌｙｍ－ｐｈｏｋｉｎｅａｃｔｉｖａｔｅｄｋｉｌｅｒｃｅｌｋｉｎｅ）活性的影响，以提高ＬＡＫ细胞杀瘤活性、减少ＩＬ－２（白细胞介素－２）用量和毒副作用。通过ＭＴＴ法测定不同浓度ＩＬ－２、不同浓度ＭＰＳ对ＬＡＫ细胞促增殖能力的影响；以细胞计数法测定不同浓度ＩＬ－２、ＭＰＳ对ＬＡＫ细胞扩增倍数的影响；应用乳酸脱氢酶法测定不同条件刺激的ＬＡＫ细胞对喉癌ＨＥＰ－２细胞杀伤活性的作用。结果证明小剂量ＩＬ－２（２×１０５Ｕ／Ｌ）诱导ＬＡＫ细胞过程中加入ＭＰＳ０．４ｍｇ／Ｌ能显著增强ＬＡＫ细胞的增殖和杀伤喉癌ＨＥＰ－２细胞的活性。其扩增倍数及杀伤活性均显著超过单纯应用ＩＬ－２（１０６Ｕ／Ｌ）诱导的ＬＡＫ细胞。通过检测不同条件下诱导的ＬＡＫ细胞表型，显示ＭＰＳ联合ＩＬ－２诱导的ＬＡＫ细胞ＣＤ２５百分率明显增高，提示ＭＰＳ可能通过促进ＬＡＫ细胞表面ＩＬ－２膜受体表达，提高ＬＡＫ细胞增殖及杀伤活性。因此，ＭＰＳ可望作为一种新型免疫调节剂用于抗肿瘤治疗     

大鼠鼻粘膜SP及CGRP逆向释放的免疫细胞化学研究

为了探讨调节肽在鼻粘膜炎症中的作用，我们用４％福尔马林滴鼻（双侧），观察大鼠鼻粘膜Ｐ物质（ｓｕｂｅｔａｎｃｅＰ，ＳＰ）及降钙素基因相关肽（ｃａｌｃｉｔｏｎｉｎｇｅｎｅｒｅｌａｔｅｄｐｅｐｔｉｄｅ，ＣＧＲＰ）免疫反应纤维的动态变化。发现每日一次、连续滴鼻５天后，鼻粘膜ＳＰ及ＣＧＲＰ纤维明显增多，以小血管壁为著；三叉神经节（ＴＧ）部分神经原胞体ＳＰ及ＣＧＲＰ免疫反应减弱。以上结果提示鼻粘膜单纯非特异性刺激可致局部ＳＰ及ＣＧＲＰ逆向释放增多。讨论了鼻粘膜ＳＰ及ＣＧＲＰ增多的临床意义。     

飘散蒿属花粉对鼻粘膜上皮细胞间粘附分子表达的影响

为探讨吸入性变应原对变应性鼻炎患者鼻粘膜上皮细胞表达细胞间粘附分子（Ｉｎｔｅｒ－ｃｅｌｕｌａｒａｄｈｅｓｉｏｎｍｏｌｅｃｕｌｅ－１，ＩＣＡＭ－１）的影响，对１１例蒿属花粉过敏的花粉症患者，采用ＲＴ－ＰＣＲ的方法，测定鼻粘膜上皮细胞ＩＣＡＭ－１的ｍＲＮＡ表达。结果显示：在蒿属花粉飘散期，８例标本全部呈阳性；在蒿属花粉非飘散期，３例标本中２例阴性，１例阳性（该例对屋内尘土皮肤试验也呈阳性反应）。由此提示：当外界环境中的特异性变应原含量增高时，花粉症患者的鼻粘膜上皮细胞表达Ｉ－ＣＡＭ－１，而无致敏原时，ＩＣＡＭ－１的表达为阴性。     

P物质及组胺受体拮抗剂对分泌性中耳炎中耳积液的影响

采用免疫组化ＡＢＣ－ＧＤＮ及图象分析技术研究分泌性中耳炎中耳粘膜Ｐ物质含量的变化与中耳积液的关系，并观察ＳＰ受体拮抗剂ｓｐａｎｔｉｄｅ及组胺Ｈ２受体阻滞剂ｃｉｍｅ－ｔｉｄｉｎ对ＭＥＥ的影响，探讨ＳＰ在ＳＯＭ中的作用。结果发现，在ＳＯＭ条件下，中耳粘膜ＳＰ含量有逐渐增多趋势，并和ＭＥＥ量呈正相关。     

两种免疫调节剂辅助LAK细胞杀伤喉癌细胞的体外研究

为探讨如何提高白细胞介素－２（ＩＬ－２）／ＬＡＫ细胞过继免疫治疗的抗肿瘤作用，应用免疫调节剂分枝杆菌多糖（Ｍｙｃｏｂａｃｔｅｒｉａｌｐｏｌｙｓａｃｃｈａｒｉｄｅｓ，ＭＰＳ）或ＣＤ３单克隆抗体协同ＩＬ－２诱导的健康人外周血淋巴细胞来源的ＬＡＫ细胞在体外扩增，测定其对喉癌ＨＥＰ－２细胞杀伤活性。实验结果：①不同免疫调节剂对ＬＡＫ细胞扩增的影响：实验组三组细胞于体外扩增均明显高于对照组（Ｐ＜０．０１）；ＭＰＳ组或ＣＤ３组细胞的扩增倍数比单纯应用ＩＬ－２组为高（Ｐ＜０．０１）；其中扩增倍数最高者为ＭＰＳ组，与ＣＤ３组相比，Ｐ＜０．０５。②不同免疫调节剂诱导的ＬＡＫ细胞对喉癌ＨＥＰ－２细胞的杀伤活性比较：在培养一周时，实验组三组对ＨＥＰ－２细胞的杀伤活性均高于对照组，以ＣＤ３组及ＭＰＳ组杀伤活性最高，其杀伤活性均为５５．５％；于培养第二周，ＣＤ３组杀伤ＨＥＰ－２细胞活性仍较高，其他实验组细胞杀伤活性均明显下降。结论：免疫调节剂ＭＰＳ和ＣＤ３单克隆抗体均可通过增强ＬＡＫ细胞体外增殖和杀瘤活性而提高肿瘤过继免疫治疗效果。     

K+ and Na+ absorption by outer sulcus epithelial cells.

Transduction of sound into nerve impulses by hair cells depends on modulation of a current carried primarily by K+ into the cell across apical transduction channels that are permeable to cations. The cochlear function thus depends on active secretion of K+ accompanied by absorption of Na+ by epithelial cells enclosing the cochlear duct. The para-sensory cells which participate in the absorption of Na+ (down to the uniquely low level of 1 mM) were previously unidentified and the existence of a para-sensory pathway which actively absorbs K+ was previously unknown. A relative short circuit current (Isc,probe, measured as the extracellular current density with a vibrating electrode) was directed into the apical side of the outer sulcus epithelium, decreased by ouabain (1 mM), an inhibitor of Na+, K(+)-ATPase, and found to depend on bath Na+ and K+ but on neither Ca2+ nor Cl-. Isc,probe was shown to be an active current by its sensitivity to ouabain. On-cell patch clamp recordings of the apical membrane of outer sulcus cells displayed a channel activity, which carried inward currents under conditions identical to those used to measure Isc,probe. Both Isc,probe and non-selective cation channels (27.4+/-0.6 ps, n = 22) in excised outside-out patches from the apical membrane were inhibited by Gd3+ (1 mM). Ics,prob was also inhibited by 5 mM lidocaine, 1 mM quinine and 500 microM amiloride but not by 10 microM amiloride. These results demonstrate that outer sulcus epithelial cells contribute to the homeostasis of endolymph by actively absorbing Na+ and K+. An entry pathway in the apical membrane was shown to be through non-selective cation channels that were sensitive to Gd3+.     

葡萄球菌肠毒素B对兔上颌窦黏膜上皮电生理和通透性的影响

目的 研究葡萄球菌肠素素B（staphylococcal enterotoxin,B,SEB) 对兔上颌窦黏膜屏障功能的影响。方法 30只兔上颌窦黏膜在手术显微镜下活体取出后装于亚森室，测定上皮的短路电流、电导和通透性以及黏膜中肿瘤坏死因子－α（tumor necrosis factor α，TNF-α）。结果 葡萄球菌肠毒素B刺激引起上颌窦黏膜上皮的短路电流、电导和对辣根过氧化物酶（horseradish peroxidase,HRP)的通透性增高。用TNF－α在体外刺激窦黏膜可引起上述同样的变化。SEB刺激窦黏膜可引起其中TNF－α含量升高。经抗TNF－α抗体预处理可以阻断SEB引起的窦黏膜上皮的上述病理改变。结论 SEB可影响上颌窦上皮层的屏幕功能，可能是通过引起其中免疫细胞的功能变化，在上颌窦炎症的发生、发展中有一定的病理学意义。     

辛芩颗粒联合H1受体拮抗剂治疗变应性鼻炎实验研究

目的探索辛芩颗粒联合H1受体拮抗剂氯雷他定对豚鼠变应性鼻炎（allergic rhinitis,AR）模型的治疗作用和对P物质（substance P,SP）及其受体（substance P receptor,SP-R）mRNA表达的影响。方法36只清洁级豚鼠随机分为4组,每组9只,分别为AR模型无干预组（A组）、辛芩颗粒干预组（B组）、H1受体拮抗剂氯雷他定于预组（C组）、辛芩颗粒联合氯雷他定干预组（D组）。各组分别给予不同药物干预后进行喷嚏、抓鼻、呼吸频率的行为学观察,同时应用免疫组化检测鼻中隔黏膜中SP分泌,逆转录聚合酶链反应测定鼻中隔黏膜中SP-R mRNA表达。结果各组间在喷嚏、抓鼻、呼吸频率、SP细胞计数及SP—R mRNA的相对表达量比较均有统计学意义（P值均〈0．05）。实验分析显示,辛芩颗粒联合氯雷他定对上述影响具有协同作用（P值均〈0．05）。结论辛芩颗粒联合氯雷他定在缓解豚鼠AR的症状,减少鼻黏膜中SP阳性细胞数量以及降低SP—R mRNA的表达优于单独用药组,并且具有协同作用。     

Kinin and histamine stimulate Cl- secretion in gerbil middle ear epithelium: connection to otitis media.

The effects of bradykinin (BK) and histamine on transepithelial ion transport in primary cultures of gerbil middle ear epithelium were investigated. Lysyl-bradykinin (lys-BK) elicited a transient increase in short-circuit current (I(sc)) when added to apical or basolateral surfaces. Lys-BK had a larger effect than BK or des-arg9-BK on both epithelial surfaces. Histamine induced a transient increase in I(sc) only when added to the basolateral surface. Mepyramine, an H1 histamine antagonist, greatly reduced the histamine-induced I(sc). The H2 and H3 histamine antagonists were both ineffective for inhibiting the I(sc) responses to histamine. Diphenylamine-2-carboxylate or 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid, Cl- channel blockers, significantly blocked the I(sc) responses to lys-BK or histamine. The Ca2+-mobilizing action of lys-BK and histamine was also investigated in single middle ear epithelial cells. BK and histamine induced an increase in the intracellular Ca2+ concentration. 1,2-Bis-(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester, a calcium chelator, greatly reduced the increase in the I(sc) responses to lys-BK or histamine. These data indicate that BK and histamine activate intracellular Ca2+-dependent mechanisms, leading to apical Cl- secretion in the cultured gerbil middle ear epithelium via B2 BK receptors and H1 histamine receptors.     

Antidromic nerve stimulation accelerates mucociliary activity in rabbit maxillary sinus

The neuropeptide substance P (SP) is released from unmyelinated C-fibres in sensory nerves by antidromic nerve stimulation. In order to study the effect on mucociliary (m.c.) activity, the isolated distal end of the maxillary nerve was stimulated electrically (8 V, pulse rate 10 Hz, 5 ms pulses) 2-4 min, while the m.c. wave frequency in the maxillary sinus was recorded by a photoelectric technique. The m.c. wave frequency increased during stimulation by 35.4 +/- 4.9% (range 22.0-62.2%). Following pretreatment with atropine the m.c. activity increased by only 20.0 +/- 1.3% (range 14.9-21.9%) indicating that part of the response was mediated via stimulation of muscarinic receptors. The cholinergic part of the response is presumably secondary to release of SP since the SP antagonist [D-Pro2, D-Trp7,9]SP abolished the effect of nerve stimulation. Sympathetic ganglionectomy did not influence the response to antidromic nerve stimulation. It is concluded that antidromic stimulation of the maxillary nerve accelerates m.c. activity through a dual mechanism, involving release of SP, or SP-like peptides, from sensory C-fibre nerve endings as well as release of acetylcholine.     

Cholinergic and C-fibre mediated mechanisms in the stimulation of mucociliary activity induced by prostaglandins and histamine

The involvement of cholinergic and C-fibre mediated mechanisms in the stimulation of mucociliary activity induced by prostaglandins and histamine was investigated in vivo in the rabbit maxillary sinus with a photoelectric technique. The prostaglandins E, (PGE,) and F2(2) alpha (PGF2 alpha) in dose of 0.1 microgram/kg and 10 micrograms/kg respectively stimulated the mucociliary activity in a biphasic fashion, with a small initial response during the first 1-2 min and a later maximum response after 3-4 min. These effects were resistant to atropine and to the SP antagonist (D-Pro2, D-Trp7,9)SP. The small initial response was blocked by pretreatment with high doses of capsaicin (13 mg i.a.), while the maximum response was unaffected. This indicates that the mucociliary responses induced by PGE, and PGF2 alpha involve capsaicin-sensitive C-fibres but that neither acetylcholine nor substance P were responsible. Histamine (50 micrograms/kg) stimulated mucociliary activity in the rabbit maxillary sinus and the effect was abolished by pretreatment with high doses of capsaicin and reduced by the SP antagonist (D-Pro2, D-Trp7,9)SP. This indicates that the histamine-induced stimulation of mucociliary activity involves capsaicin-sensitive C-fibres and that the effect might be mediated by substance P.     

组胺H3受体拮抗剂对变应性鼻炎大鼠的作用

目的通过变应性鼻炎（AR）动物模型,观察组胺H3受体拮抗剂噻普酰胺（thioperamide）对AR病理生理过程的影响,从而探索组胺H3受体拮抗剂在AR中是否存在潜在的治疗作用。方法 60只Wistar大鼠以随机数字表法分为5组,每组12只,分别为正常对照组（NC组）、AR模型未干预组（AR组）、组胺H1受体拮抗剂氯雷他定处理组（HR1组）、组胺H3受体拮抗剂处理组（HR3组）及组胺H1、H3受体拮抗剂联合处理组（HR1＋3组）。卵清蛋白法建立ARWistar大鼠模型,对照组胺H1受体拮抗剂氯雷他定、组胺H3受体拮抗剂thioperamide处理大鼠及联合干预组与未处理组大鼠血清及鼻腔灌洗液中细胞因子及炎症介质方面的变化。结果与AR组相比,HR1组大鼠,血清IgE、IL-4及鼻腔灌洗液中嗜酸细胞活化趋化因子水平显著下降,血清INF-γ水平升高,HR3组无显著变化,HR1＋3组与HR1组相比,各指标亦无显著改变。结论组胺H3受体拮抗剂thioperamide不能缓解AR炎症状态,组胺H3受体拮抗剂与组胺H1受体拮抗剂在AR治疗方面未发现有协同或叠加作用。