Identification of immunoreactive substance P in human and other mammalian endothelial cells

doi:10.1016/0196-9781(89)90175-7

Peptides

Volume 10, Issue 5, September–October 1989, Pages 957-962

https://doi.org/10.1016/0196-9781(89)90175-7 Get rights and content

Abstract

Endothelial cells release both vasodilatory (e.g., PGI₂, EDRF, oxygen radicals) and vasoconstrictor (e.g., EDCF) substances which modify vascular tone and contractility. We report the existence of the vasodilatory tachykinin substance P within endothelial cell scraping from human, rat and dog thoracic aorta and human pial arteries with values ranging from $1.0±0.1$ (rat aorta) to $1.9±0.5$ (dog aorta) fmol/mg protein. The immunoreactive component eluted with a retention time identical to that of radiolabelled substance P when analyzed by high performance liquid chromatography combined with radioimmunoassay. Cultured endothelial cells from bovine cerebral microvessels contained measurable levels of substance P in passages 3–8, suggesting the likelihood that these cells synthesize substance P. However, the level of gene expression must be low since efforts to demonstrate the presence of preprotachykinin mRNA by Northern blot analysis of dog and rat aortic endothelial cell RNA or by RNase protection analysis of rat aortic endothelial cell RNA were not successful.

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Cited by (92)

The two-faced effects of nerves and neuropeptides in corneal diseases
2022, Progress in Retinal and Eye Research
Citation Excerpt :
It is encoded by the TAC1 gene, which is located on the human chromosome 7. SP is mainly released by sensory terminals, although immune (Suvas, 2017), epithelial (Watanabe et al., 2002), endothelial (Linnik and Moskowitz, 1989), and bone marrow stem cells (Li et al., 2000) can also produce it. SP exerts its biological activities by binding to tachykinin receptors (NK1R, NK2R, and NK3R) expressed in both neuronal and non-neuronal cells (Khawaja and Rogers, 1996).
Corneal nerves are instrumental to maintain cornea integrity through regulation of key physiological functions such as tear secretion, blink reflex, and neuropeptide turnover. Corneal nerve injury/stimulation can follow many insults including mechanical/chemical trauma, infections and surgeries. Nerve disruption initiates a process named neurogenic inflammation which leads to edema, pain, and recruitment and activation of leukocytes. Interestingly, leukocyte influx in the cornea can further damage nerves by releasing inflammatory mediators-including neuropeptides. The clinical outcome of neuroinflammation can be beneficial or detrimental to corneal integrity. On one side, it ensures prompt wound healing and prevents infections. On the other, prolonged and/or deranged neuroinflammation can permanently disrupt corneal integrity and impair vision.
The cornea is an ideal site to study peripheral neuroinflammation and neurogenic inflammation since it receives the highest density of sensory nerves of the entire body.
We will review the corneal nerve anatomy and neurochemistry, discuss the beneficial and detrimental effects of neurogenic inflammation in corneal wound healing, inflammatory processes, and pain. We will also examine the emerging remote impact of corneal nerve disruption on the trigeminal ganglion and the brain, highlighting the key role of neuropeptide Substance P. Finally, we will discuss the clinical relevance of such neuroinflammatory network in the context of severe and highly prevalent ocular diseases, including potential treatments.
Association between serum substance P levels and mortality in patients with severe sepsis
2015, Journal of Critical Care
Citation Excerpt :
The TAC1 gene encodes substance P (SP), a member of the tachykinin family largely considered a peptide of neuronal origin [3]. However, SP and other tachykinins are also widely distributed in nonneuronal tissues [4–7] where they mediate multiple homeostatic functions, including nociception and neurogenic inflammation [3,8]. The effects of SP are mainly mediated by the NK1receptor (NK1R), a G-protein–coupled receptor that is widely expressed in many tissues and cells [7,9].
Substance P (SP) is a peptide of the tachykinins family involved in the inflammatory response. Circulating SP levels have been assessed in septic patients in 2 previous studies with a small number of subjects (61 and 42 patients, respectively), and there were no significant differences in SP levels at the moment of sepsis diagnosis between surviving and nonsurviving patients. The main goal of this study was to determine a possible relationship between serum SP levels and patient outcome in the largest cohort of severe septic patients analyzed so far.
We performed an observational, prospective, multicenter study in 6 Spanish intensive care units. Serum SP levels were measured at the moment of severe sepsis diagnosis in 238 patients. The end point of the study was 30-day mortality.
We found that surviving septic patients (n = 153) showed higher serum SP levels than did nonsurvivors (n = 85). Multiple logistic regression analysis showed that serum SP levels higher than 350 pg/mL were associated with survival at 30 days (odds ratio, 0.43; 95% confidence interval, 0.24-0.77; P = .005) after controlling for serum lactic acid levels and Sepsis-related Organ Failure Assessment score.
The major new finding of our study was that serum SP levels were associated with mortality in severe septic patients.
Morphology and neurochemistry of rabbit iris innervation
2015, Experimental Eye Research
Citation Excerpt :
The presence of SP-positive cell-like structures in the both sides of the iris is a new finding. In addition to SP be recognized as a neuropeptide in sensory neurons, SP gene expression has been reported in non-neural cells, such as human fibroblasts, keratinocytes, endothelial cells, lymphocytes and platelets (Bae et al., 2002; Jones et al., 2008; Lai et al., 1998; Linnik and Moscowitz, 1989). Our previous study showed that rabbit corneal epithelial cells express SP after experimental surgery (Cortina et al., 2012).
The aim of this study was to map the entire nerve architecture and sensory neuropeptide content of the rabbit iris. Irises from New Zealand rabbits were stained with antibodies against neuronal-class βIII-tubulin, calcitonin gene-related peptide (CGRP) and substance P (SP), and whole-mount images were acquired to build a two-dimensional view of the iridal nerve architecture. After taking images in time-lapse mode, we observed thick nerves running in the iris stroma close to the anterior epithelia, forming four to five stromal nerve rings from the iris periphery to the pupillary margin and sub-branches that connected with each other, constituting the stromal nerve plexus. In the anterior side, fine divisions derivated from the stromal nerves, forming a nerve network-like structure to innervate the superficial anterior border layer, with the pupillary margin having the densest innervation. In the posterior side, the nerve bundles ran along with the pupil dilator muscle in a radial pattern. The morphology of the iris nerves on both sides changed with pupil size. To obtain the relative content of the neuropeptides in the iris, the specimens were double stained with βIII-tubulin and CGRP or SP antibodies. Relative nerve fiber densities for each fiber population were assessed quantitatively by computer-assisted analysis. On the anterior side, CGRP-positive nerve fibers constituted about 61%, while SP-positive nerves constitute about 30.5%, of the total nerve content, which was expressed as βIII tubulin-positive fibers. In addition, in the anterior stroma of the collarette region, there were non-neuronal cells that were positive for SP. On the posterior side, CGRP-positive nerve fibers were about 69% of total nerve content, while SP constituted only up to 20%. Similarly, in the trigeminal ganglia (TG), the number of CGRP-positive neurons significantly outnumbered those that were positive for SP. Also, all the SP-positive neurons were labeled with CGRP. This is the first study to provide a two-dimensional whole mount and a cross-sectional view of the entire iris nerve architecture. Considering the anatomical location, the high expression of CGRP and SP suggests that these neuropeptides may play a role in the pathogenesis of anterior uveitis, glaucoma, cataracts and chronic ocular pain.
Immunolocalization of NK-1 Receptor and Substance P in Human Normal Placenta
2010, Placenta
Citation Excerpt :
Traditionally it has been classified as a neurotransmitter exerting its effects on the periphery only by being released from nerve endings. However, SP has been demonstrated in murine and human non-neuronal cell types, including endothelial cell [2,3] macrophages and monocytes [4], eosinophils [5], lymphocytes [6], and Leydig cells [7]. This suggests that it could not only act as a neurotransmitter but also as a functional regulator in an autocrine or paracrine manner.
A role for Substance P (SP) in human placenta is not known, although is possible that regulates placental physiology through the Neurokinin (NK)-1 receptor. Ten human normal placenta tissues were studied by immunohistochemistry to demonstrate the localization of NK-1 receptor and SP. An immunostaining pattern for NK-1 receptor and SP was observed in the endothelium and myocytes of fetal blood vessels, decidua and trophoblast. The SP is located in both the cytoplasm and the nucleus whereas NK-1 receptor in cytoplasmic. These findings reported here for the fist time, suggest a role for the SP and NK-1 receptor in the placental physiology.
Differentiation of neuronal from non-neuronal Substance P
2009, Regulatory Peptides
Substance P (SP) originally found as a neuropeptide in capsaicin-sensitive sensory neurons, had more recently been identified in non-neuronal cells, especially under pathological conditions. Neuronal and non-neuronal SP may perform distinct functions. A simple technique to differentiate different SP sources is currently unavailable. Herein, we describe a two-step sequential acetic acid extraction to differentiate SP source. The efficiency of this two-step extraction in differentiating SP in capsaicin-sensitive neurons was verified by using capsaicin as a tool to deplete SP in sensory neurons. Specifically, Balb-c mice were treated with high dose capsaicin (200 mg/kg). Skin was removed two weeks after treatment. In a separate experiment, lung and skin tissues from control animals (untreated) were incubated in-vitro with capsaicin, and sequential acetic acid extraction was performed. Following capsaicin treatment, both in-vivo and in-vitro, SP recovered in first extraction decreased significantly in lung and skin. Lastly, presence of capsaicin solvent (10% methanol and 10% Tween 80) or protease inhibitor cocktail in solution altered SP EIA test, yielding false positive results. These results demonstrated that SP in capsaicin sensitive sensory neurons was extracted in initial extraction of 15 min while non-neuronal SP was present in second extraction. Because SP in non-neuronal tissues may possibly be more important in pathological conditions, this technique could be useful in determining effects of various treatments on neuronal and non-neuronal SP levels and their consequences.
In vitro substance P-dependent induction of bone marrow cells in common (CD10) acute lymphoblastic leukaemia
2008, Leukemia Research
Citation Excerpt :
As compared to mature lymphocytes, lymphoblasts carry around 3–4-fold higher amounts of receptors for SP [12]. Apart from lymphocytes, receptors for SP also can be noted on monocytes [13], endothelial cells [14], fibroblasts [15] and hematopoietic stem cells [16]. Substance P augments proliferative activity of human and mouse lymphocytes T [17,18], human smooth muscle cells [19], mouse fibroblasts [20], fibroblasts of human skin [15], smooth muscle fibers of arterial walls [21], human synovial cells [22] and human cells forming colonies of granulocytes and monocytes or of erythrocytes [23].
The aim of the present research was to investigate the possible in vitro stimulatory effect of substance P (SP) on blasts induction in childhood common acute lymphoblastic leukaemia (ALL). Bone marrow aspirates were incubated with SP receptor agonist or antagonist (spantide) and subsequently assayed for the presence of human interleukin (IL)-1b using ELISA kit.
Blast cells incubated with SP receptor agonist were found to result in a significant increase of IL-1b concentration while incubated with spantide resulted in control levels of IL-1b. These findings suggest the novel possible role of SP in blasts proliferation in childhood ALL of common (CD10) origin.

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ArticleIdentification of immunoreactive substance P in human and other mammalian endothelial cells

Abstract

J. Biol. Chem.

J. Chromatogr.

J. Biol. Chem.

FEBS Lett.

Neuroscience

Neuroscience

Mol. Brain Res.

Biochim. Biophys. Acta

Tissue Cell