瞬时感受器电位香草酸受体1在变应性鼻炎鼻黏膜中的作用机制

瞬时感受器电位香草酸受体1（TRPV1）通道是一种非选择性阳离子通道,其可以被辣椒素、高温等多种刺激因素激活,参与多种生理及病理过程。最新研究证实,TRPV1离子通道是神经 免疫调控网络机制的核心要素,在变应性鼻炎鼻黏膜中发挥重要作用。我们通过对TRPV1经神经 免疫调控网络调节神经肽,进而影响鼻部症状的研究进展做简要综述,期望为临床治疗变应性鼻炎提供理论依据。     

Origin and peptide content of nerve fibers in the nasal mucosa of rats

Injection of the retrograde neuronal tracer True blue into the anterior-lateral part of the nasal mucosa of rats labeled nerve cell bodies in the superior cervical ganglion, the sphenopalatine ganglion, the otic ganglion and the trigeminal ganglion on the ipsilateral side. In the superior cervical ganglion, the sphenopalatine ganglion and the trigeminal ganglion on the contralateral side, very few nerve cell bodies were labeled, indicating that these ganglia provide minor contributions only. The number of labeled cell bodies indicates that the superior cervical ganglion, the sphenopalatine ganglion and the trigeminal ganglion contribute most to the innervation of the nose, while the contribution from the otic ganglion is minor. Cell bodies in the superior cervical ganglion harbored noradrenaline (NA) or NA/neuropeptide Y (NPY); in the sphenopalatine ganglion vasoactive intestinal peptide (VIP) or VIP/NPY; in the otic ganglion VIP, VIP/NPY or VIP/substance P (SP) and in the trigeminal ganglion calcitonin gene-related peptide (CGRP) or CGRP/SP. The results from denervations and tracer experiments suggest that all NA-containing and the majority of NPY-containing fibers in the nasal mucosa are derived from the superior cervical ganglion (sympathetic nerve supply). VIP- and VIP/NPY-containing fibers originate from the sphenopalatine and otic ganglia (parasympathetic nerve supply). Nerve fibers containing CGRP and CGRP/SP emanate from the trigeminal ganglion (sensory nerve supply).     

A ganglion probably belonging to the N. terminalis system in the nasal mucosa of the mouse

Summary In mouse, rat and hamster there occurs in the nasal mucosa and far forward in the nose a somewhat diffuse ganglion of placodal origin. It persists throughout life. The identity of the associated nerve has not yet been established, but N. terminalis and N. ethmoidalis anterior may both be involved. Comparable structures seem to be present in some mammals, but not in others. The function of the ganglion is unknown.     

Vascular effects of slow reacting substance (SRS) in the cat nasal mucosa and the appearance of a SRS-like principle in cat nasal secretion on nerve stimulation

Stimulation of parasympathetic secreto-vasomotor nerve fibres in cat nasal mucosa produced nasal secretions with biological activity similar to cat paw SRS. The biologically active principle was purified by ethanol extraction, Amberlite XAD-2 and anion exchange chromatography. It had elution characteristics common with cat paw SRS but different to those of prostaglandin F_2α. The biological activity on the guinea pig ileum was antagonized by FPL 55712. In the cat intraarterial administration of cat paw SRS resulted in a dose-dependent but modest increase in nasal blood flow whereas local blood content was not affected. Provided SRS has similar effects on human nasal blood flow the importance for SRS as a mediator for nasal congestion in allergic rhinitis or parasympathetically induced vasomotor rhinitis seems less likely.     

CCR7-independent transport of skin antigens occurs in the dermis

Under homeostatic conditions, skin DCs migrate to regional LNs transporting self-antigens (self-Ags). The transport of self-Ags is considered to be critical for maintaining peripheral tolerance. Although the chemokine receptor CCR7 potently induces the migration of skin DCs to regional LNs, Ccr7(-/-) (Ccr7-KO) mice do not show skin auto-immune diseases. To resolve this inconsistency, we examined Ccr7-KO epidermis- or dermis-hyperpigmented transgenic (Tg) mice, in which the transport of skin self-Ags is traceable by melanin granules (MGs). Under CCR7-deficient conditions, the transport of epidermal MGs to regional LNs was impaired at 7 weeks of age. However, epidermal MGs could be transported when they had accumulated in the dermis. Ccr7-KO-dermis-pigmented Tg mice confirmed the presence of CCR7-independent transport from the dermis. Compared with WT-dermis-pigmented Tg mice, the amount of transported melanin and number of MG-laden CD11c(+) cells were both approximately 40% of the WT levels, while the number of MG-laden CD205(+) or CD207(+) cells decreased to about 10% in skin regional LNs of Ccr7-KO-dermis-pigmented Tg mice. Cell sorting highlighted the involvement of CD11c(+) cells in the CCR7-independent transport. Here, we show that CCR7-independent transport of skin self-Ags occurs in the dermis. This system might contribute to the continuous transport of self-Ags, and maintain peripheral tolerance.     

Choline transport and its osmotic regulation in renal cells derived from the rabbit outer medullary thick ascending limb of Henle

 Organic osmolytes such as betaine and glycerophosphorylcholine (GPC) are of major importance concerning volume regulation of inner and outer medullary epithelial cells. Recently we demonstrated that the intracellular betaine content in rabbit kidney cells derived from the outer medullary thick ascending limb of Henle’s loop (TALH) is osmotically regulated by betaine synthesis. In this context it was our purpose to characterize the uptake of choline, a precursor of betaine and GPC. We found TALH cells to possess a specific choline transport system with a maximum velocity (V _max) of 71 ± 12 pmol ·μl^–1 cell water · min^–1 and an apparent affinity (K _m) of 155 ± 19 μmol · l^–1. The uptake of choline was sodium independent and not electrogenic, but it was significantly reduced by the removement of chloride from the incubation medium. After long-term adaptation of TALH cells to a hyperosmotic medium (600 mosmol · l^–1, osmolarity adjusted with NaCl or urea) a significant higher choline uptake rate was observed (V _max: 166 ± 9 (NaCl), 96 ± 12 (urea) pmol ·μl^–1 cell water · min^–1). Our results suggest that the uptake of choline is due to higher intracellular requirements of choline under hypertonic conditions. Finally, an increase in the V _max of the choline transport system may enable sufficient synthesis of betaine and GPC. Received: 7 April 1997 / Received after revision: 2 June 1997 / Accepted: 3 June 1997     

鼻内针刺对兔变态反应性鼻炎模型鼻黏膜病理学的作用及瞬时感受器电位香草酸受体1-P物质轴相关的调控机制#br#

目的 通过鼻内针刺明确鼻黏膜的病理学变化并通过瞬时感受器电位香草酸受体1(TRPV1)-P物质(SP)轴在实验性变应性鼻炎鼻黏膜中分布及表达情况,探讨其可能的信号传导机制。 方法 采用卵清蛋白及氢氧化铝凝胶方法建立变应性鼻炎动物模型。25只新西兰大耳兔随机分成正常组、模型组、假针刺组、针刺外迎香组和鼻内针刺组,每组各5只。观察鼻内针刺对动物行为学、鼻黏膜病理学及动脉血中嗜酸性粒细胞(EOS)计数和IgE含量影响,采用HE染色法明确鼻黏膜EOS的分布,采用免疫组织化学方法观察鼻黏膜及TRPV1及SP的分布和表达情况。 结果 行为学评分结果发现,与模型组相比,假针刺组行为学计分下降不明显,针刺外迎香组和鼻内针刺组行为学计分有下降趋势,鼻内针刺组下降程度更显著。血常规中EOS计数结果显示,模型组与正常组相比,EOS计数稍增多但差别无显著性（P>0.05）;鼻内针刺组与模型组相比,EOS计数有减少趋势,差别也无显著性（P>0.05）。ELISA法检测IgE发现,与正常组相比,模型组血清中IgE含量明显增多（P<0.05）;与模型组相比,假针刺组血清中IgE含量未见明显变化（P>0.05）,针刺外迎香组和鼻内针刺组血清IgE含量有下降趋势（P<0.05）,但鼻内针刺组血清IgE含量较针刺外迎香组下降更为明显,两组差异无统计学意义（P>0.05）。HE染色结果显示,与正常组相比,模型组和假针刺组EOS浸润明显增多, 差异有统计学意义（P<0.05）;与模型组相比,鼻内针刺组和外迎香组EOS浸润程度减轻,差异有统计学意义（P<0.05）。免疫组织化学结果显示,与正常组相比,模型组和假针刺组鼻黏膜TRPV1及SP表达增多,差异有统计学意义（P<0.05）;与模型组相比,鼻内针刺组和外迎香组TRPV1、SP表达明显减弱,差异有统计学意义（P<0.05）。 结论 鼻内针刺可以通过减少嗜酸性粒细胞在鼻黏膜的趋化及血清中IgE含量来缓解变应性鼻炎模型的鼻部症状。     

Corticosteroids stimulate the amphibious behavior in mudskipper: potential role of mineralocorticoid receptors in teleost fish

It has long been held that cortisol, a glucocorticoid in many vertebrates, carries out both glucocorticoid and mineralocorticoid actions in teleost fish. However, 11-deoxycorticosterone (DOC) has been identified as a specific endogenous ligand for the teleostean mineralocorticoid receptor (MR). Furthermore, the expressions of MR mRNA are modest in the osmoregulatory organs, but considerably higher in the brain of most teleosts. These recent findings suggest that the mineralocorticoid system (DOC/MR) may carry out some behavioral functions in fish. To test this possibility, we examined the effects of cortisol and DOC administration in the amphibious behavior in mudskipper (Periophthalmus modestus) in vivo. It was found that mudskippers remained in the water for an increased period of time when they were immersed into 5 μM DOC or cortisol for 8 h. Additionally, an exposure to 25 μM DOC for 4 to 8 h caused a decreased migratory frequency of mudskippers to the water, reflected a tendency to remain in the water. It was further observed that after 8 h of intracerebroventricular (ICV) injection with 0.3 pmol DOC or cortisol the staying period in the water increased in fish. The migratory frequency was decreased after ICV DOC injection which indicated that fishes stayed in the water. Concurrent ICV injections of cortisol with RU486 [a specific glucocorticoid-receptor (GR) antagonist] inhibited only the partial effects of cortisol. Together with no changes in the plasma DOC concentrations under terrestrial conditions, these results indicate the involvement of brain MRs as cortisol receptors in the preference for an aquatic habitat of mudskippers. Although the role of GR signaling cannot be excluded in the aquatic preference, our data further suggest that the MR may play an important role in the brain dependent behaviors of teleost fish.     

Basolateral transport of glutarate in proximal S2 segments of rabbit kidney: kinetics of the uptake process and effect of activators of protein kinase A and C

 The kinetics of tubular glutarate uptake, the coupling of glutarate to p-aminohippurate (PAH) transport and the effect of activators of protein kinase A and C on glutarate uptake were studied using isolated S₂ segments of proximal tubules microdissected from rabbit kidneys without the use of enzymatic agents. Because the tubules were not perfused, and hence were collapsed, the tubular uptake of [¹⁴C]glutarate reflects transport across the basolateral cell membrane. To obtain uptake rates most closely related to initial transport rates, 30 s glutarate uptake measurements were performed. In a first set of experiments it could be shown that preloading proximal S₂ segments with glutarate (10^–3 M) stimulated [³H]PAH uptake indicating that glutarate may be a substrate of the PAH /dicarboxylate exchanger. The kinetic data revealed a K _m value of 0.62 mM and a V _max value of 84.1 pmol nl^–1min^–1 for tubular [¹⁴C]glutarate uptake across the basolateral cell membrane. In contrast to basolateral PAH transport (previous studies from this laboratory), tubular 30 s [¹⁴C]glutarate uptake was not affected by either the phorbol ester phorbol 12-myristate 13-acetate (PMA, 10^–7 M), an activator of protein kinase C, or by the membrane-permeant analogues of cAMP, dibutyryl cyclic AMP (db-cAMP, 10^–4 M) and 8-bromoadenosine 3′,5′-cyclic monophosphate (Br-cAMP, 10^–4 M). The results indicate that the protein kinases A and C have no function in the regulation of the renal basolateral dicarboxylate transporter. This finding agrees well with the structural feature of the recently cloned rabbit renal dicarboxylate transporter which does not contain any putative phosphorylation sites for protein kinase C or cAMP-dependent kinase. Received: 30 December 1997 / Received after revision and accepted: 25 February 1998     

The lack of effect of benzalkonium chloride on the cilia of the nasal mucosa in patients with perennial allergic rhinitis: a combined functional, light, scanning and transmission electron microscopy study

Background Controversy still exists about the effect of 0.02% benzalkonium chloride (BKC). a preservative in many nasal sprays, on human nasal epithelium in vivo. Objective To determine the safety of BKC by assessing its effect on the function and morphology of cilia of human nasal epithelium. Methods A single-centre, double-blind nasal biopsy study in 22 patients with perennial allergic rhinitis, receiving fluticasone propionate aqueous nasal spray (FPANS) containing BKC. BKC plus placebo or placebo alone for 6 weeks. Before, at two weekly intervals during treatment and 2 weeks after treatment ceased an indigocarmine saccharine transport time (ICST) was performed. Results ICST results did not significantly vary between the groups. There was no statistical relationship between the number of ciliated cells present and the treatment the patients received. Scanning and transmission electron microscopy examination showed no effects of BKC. Conclusion Despite reports of its ciliostatic effects in vitro. BKC did not have such an effect when it was applied for 6 weeks (with/without fluticasone propionale) to the nasal mucosa of perennial allergic rhinitis patients in vivo.     

Hormonal stimulation of Ca2+ and Mg2+ transport in the cortical thick ascending limb of Henle's loop of the mouse: evidence for a change in the paracellular pathway permeability

Recent studies from our laboratory have shown that in the cortical thick ascending limb of Henle's loop of the mouse (cTAL) Ca²⁺ and Mg²⁺ are reabsorbed passively, via the paracellular shunt pathway. In the present study, cellular mechanisms responsible for the hormone-stimulated Ca²⁺ and Mg²⁺ transport were investigated. Transepithelial voltages (PD_te) and transepithelial ion net fluxes (J _Na, J _Cl, J _K, J _Ca, J _Mg) were measured in isolated perfused mouse cTAL segments. Whether parathyroid hormone (PTH) is able to stimulate Ca²⁺ and Mg²⁺ reabsorption when active NaCl reabsorption, and thus PD_te, is abolished by luminal furosemide was first tested. With symmetrical lumen and bath Ringer's solutions, no Ca²⁺ and Mg²⁺ net transport was detectable, either in the absence or in the presence of PTH. In the presence of luminal furosemide and a chemically imposed lumen-to-bath directed NaCl gradient, which generates a lumen-negative PD_te, PTH slightly but significantly increased Ca²⁺ and Mg²⁺ net secretion. In the presence of luminal furosemide and a chemically imposed bath-to-lumen-directed NaCl gradient, which generates a lumen-positive PD_te, PTH slightly but significantly increased Ca²⁺ and Mg²⁺ net reabsorption. In view of the observed small effect of PTH on passive Ca²⁺ and Mg²⁺ movement, a possible interference of furosemide with the hormonal response was considered. To investigate this possibility, Ca²⁺ and Mg²⁺ transport was first stimulated with PTH in tubules under control conditions. Then active NaCl reabsorption was abolished by furosemide and the effect of PTH on J _Ca and J _Mg measured. In the absence of PD_te and under symmetrical conditions, no Ca²⁺ and Mg²⁺ transport was detectable, either in the presence or absence of PTH. In the presence of a bath-to-lumen-directed NaCl gradient, Ca²⁺ and Mg²⁺ reabsorption was significantly higher in the presence than in the absence of PTH. Finally, when active NaCl transport was not inhibited by furosemide, but reduced by a bath-to-lumen-directed NaCl gradient, PTH strongly increased J _Ca and J _Mg, whereas only a small increase in PD_te was noted. In conclusion, these data suggest that PTH exerts a dual action on Ca²⁺ and Mg²⁺ transport in the mouse cTAL by increasing the transepithelial driving force for Ca²⁺ and Mg²⁺ reabsorption through hormone-mediated PD_te alterations and by modifying the passive permeability for Ca²⁺ and Mg²⁺ of the epithelium, very probably at the level of the paracellular shunt pathway.