RESUMEN
Esophageal cancers (EC) are highly aggressive tumors, commonly presented in a locally advanced stage with a poor prognosis and survival. Up to 50% of the patients are eligible for treatment with curative intent and receive the standard treatment with neoadjuvant chemoradiotherapy (nCRT) and surgery. Currently, pathologic complete response to nCRT is 20-30%, with a partial or no response in about 50% and 20%, respectively. EC recurrences occur frequently even after successful anti-cancer treatment, suggesting high aggressiveness with increased metastatic potential. A tumor sub-population of so-called cancer stem cells (CSCs), is known to display a high metastatic potential and resistance to conventional anti-cancer therapy, hereby being responsible for the unbeneficial clinical features. In this review, a concise overview will be given of the current literature on esophageal CSCs and related metastases. Esophageal CSC markers will be discussed followed by the pathways that initiate and sustain these cells. In addition, the cellular processes, epithelial-mesenchymal transition (EMT), hypoxia and autophagy, known to contribute to cancer stemness and metastasis will be explained. Finally, potential options for treatment also related to cancer genome atlas (TCGA) data on EC will be discussed.
Asunto(s)
Transición Epitelial-Mesenquimal/efectos de los fármacos , Neoplasias Esofágicas/tratamiento farmacológico , Terapia Neoadyuvante/métodos , Células Madre Neoplásicas/efectos de los fármacos , Autofagia/efectos de los fármacos , Hipoxia de la Célula/efectos de los fármacos , Neoplasias Esofágicas/genética , Neoplasias Esofágicas/patología , Humanos , Metástasis de la NeoplasiaRESUMEN
Radiation-induced hyposalivation is still a major problem after radiotherapy for head and neck cancer. Current and promising new thoughts to reduce or salvage radiation damage to salivary gland tissue are explored. The main cause underlying radiation-induced hyposalivation is a lack of functional saliva-producing acinar cells resulting from radiation-induced stem cell sterilization. Current methods to prevent that damage are radiation techniques to reduce radiation-injury to salivary gland tissue, surgical techniques to relocate salivary glands to a region receiving a lower cumulative radiation dose, and techniques to make salivary gland cells more resistant to radiation injury. These preventive techniques cannot be applied in all cases, also reduce tumor sensitivity, or do not result in a sufficient amelioration of the dryness-related complaints. Therefore, alternative methods on techniques to salvage salivary glands that are damaged by radiation are explored with promising results, such as stem cell therapies and gene transfer techniques to allow the radiation-injured salivary gland tissue to secrete water.
Asunto(s)
Traumatismos por Radiación/prevención & control , Glándulas Salivales/efectos de la radiación , Neoplasias de Cabeza y Cuello/radioterapia , Humanos , Terapia de Protones , Traumatismos por Radiación/terapia , Protectores contra Radiación/uso terapéutico , Radioterapia de Intensidad Modulada/efectos adversos , Xerostomía/etiología , Xerostomía/terapiaRESUMEN
BACKGROUND & AIM: Irradiation of the salivary glands during head and neck cancer treatment induces cellular senescence in response to DNA damage and contributes to radiation-induced hyposalivation by affecting the salivary gland stem/progenitor cell (SGSC) niche. Cellular senescence, such as that induced by radiation, is a state of cell-cycle arrest, accompanied by an altered pro-inflammatory secretome known as the senescence-associated secretory phenotype (SASP) with potential detrimental effects on the surrounding microenvironment. We hypothesized that the pro-regenerative properties of mesenchymal stem cells (MSCs) may attenuate cellular senescence post-irradiation. Therefore, here we evaluated the effects of adipose-derived MSCs (ADSCs) on the radiation-induced response of salivary gland organoids (SGOs). METHODS: Proteomic analyses to identify soluble mediators released by ADSCs co-cultured with SGOS revealed secretion of hepatocyte growth factor (HGF) in ADSCs, suggesting a possible role in the stem cell crosstalk. Next, the effect of recombinant HGF in the culture media of ex vivo grown salivary gland cells was tested in 2D monolayers and 3D organoid models. RESULTS: Treatment with HGF robustly increased salivary gland cell proliferation. Importantly, HGF supplementation post-irradiation enhanced proliferation at lower doses of radiation (0, 3, 7 Gy), but not at higher doses (10, 14 Gy) where most cells stained positive for senescence-associated beta-galactosidase. Furthermore, HGF had no effect on the senescence-associated secretory phenotype (SASP) of irradiated SGOs, suggesting there may be compensatory proliferation by cell-division competent cells instead of a reversal of cellular senescence after irradiation. CONCLUSION: ADSCs may positively influence radiation recovery through HGF secretion and can promote the ex vivo expansion of salivary gland stem/progenitor cells to enhance the effects of co-transplanted SGSC.
Asunto(s)
Factor de Crecimiento de Hepatocito , Células Madre Mesenquimatosas , Humanos , Factor de Crecimiento de Hepatocito/farmacología , Proteómica , Glándulas Salivales , Senescencia Celular/efectos de la radiación , Proliferación CelularRESUMEN
BACKGROUND: Pulmonary arterial hypertension (PAH) is a commonly fatal pulmonary vascular disease that is often diagnosed late and is characterised by a progressive rise in pulmonary vascular resistance resulting from typical vascular remodelling. Recent data suggest that vascular damage plays an important role in the development of radiation-induced pulmonary toxicity. Therefore, the authors investigated whether irradiation of the lung also induces pulmonary hypertension. METHODS: Different sub-volumes of the rat lung were irradiated with protons known to induce different levels of pulmonary vascular damage. RESULTS: Early loss of endothelial cells and vascular oedema were observed in the irradiation field and in shielded parts of the lung, even before the onset of clinical symptoms. 8 weeks after irradiation, irradiated volume-dependent vascular remodelling was observed, correlating perfectly with pulmonary artery pressure, right ventricle hypertrophy and pulmonary dysfunction. CONCLUSIONS: The findings indicate that partial lung irradiation induces pulmonary vascular remodelling resulting from acute pulmonary endothelial cell loss and consequential pulmonary hypertension. Moreover, the close resemblance of the observed vascular remodelling with vascular lesions in PAH makes partial lung irradiation a promising new model for studying PAH.
Asunto(s)
Hipertensión Pulmonar/patología , Pulmón/efectos de la radiación , Arteria Pulmonar/efectos de la radiación , Análisis de Varianza , Animales , Edema/patología , Endotelio Vascular/efectos de la radiación , Hemodinámica , Modelos Lineales , Pulmón/patología , Masculino , Protones , Traumatismos por Radiación/patología , Ratas , Ratas WistarRESUMEN
Hyposalivation underlying xerostomia after radiotherapy is still a major problem in the treatment of head and neck cancer. Stem cell therapy may provide a means to reduce radiation-induced hyposalivation and improve the quality of life of patients. This review discusses the current status in salivary gland stem cell research with respect to their potential to attenuate salivary gland dysfunction. Knowledge on the embryonic development, homeostasis and regeneration after atrophy of the salivary glands has provided important knowledge on the location of the salivary gland as well as on the factors that influence proliferation and differentiation. This knowledge has helped to locate, isolate and characterize cell populations that contain the salivary gland stem cell, although the exact tissue stem cell is still unidentified. The role that stem/progenitor cells play in the response to radiation and the factors that can influence stem/progenitor induced proliferation and differentiation are discussed. Finally, the mobilization and transplantation of stem cells and supportive cells and their potential to attenuate radiation-induced salivary gland damage are discussed. Based on the major advances made in the field of stem cell research, stem cell-based therapy has great potential to allow prevention or treatment of radiation-induced hyposalivation.
Asunto(s)
Traumatismos por Radiación/cirugía , Glándulas Salivales/efectos de la radiación , Trasplante de Células Madre , Xerostomía/cirugía , Células Madre Adultas/fisiología , Células Madre Adultas/trasplante , Diferenciación Celular/fisiología , Proliferación Celular , Neoplasias de Cabeza y Cuello/radioterapia , Humanos , Células Madre Mesenquimatosas/fisiología , Regeneración/fisiología , Glándulas Salivales/citología , Xerostomía/etiologíaRESUMEN
ß-Adrenergic signaling blockade is a mainstay of hypertension management. One percent of patients taking ß-blockers develop reduced salivary gland (SG) function. Here we investigate the role of SG progenitor cells in ß-blocker-induced hyposalivation, using human SG organoid cultures (SGOs). Compared with control SGs, initial low SG progenitor cell yield from patients taking ß-blockers was observed. When passaged, these SGOs recovered self-renewal and upregulated Notch pathway expression. Notch signaling was downregulated in situ in ß-adrenergic receptor-expressing luminal intercalated duct (ID) cells of patients taking ß-blockers. Control SGOs treated with ß-adrenergic agonist isoproterenol demonstrated increased proportion of luminal ID SGO cells with active Notch signaling. Control SGOs exposed to isoproterenol differentiated into more mature SGOs (mSGOs) expressing markers of acinar cells. We propose that ß-blocker-induced Notch signaling reduction in luminal ID cells hampers their ability to proliferate and differentiate into acinar cells, inducing a persistent hyposalivation in some patients taking ß-blocking medication.
Asunto(s)
Receptores Adrenérgicos/metabolismo , Receptores Notch/metabolismo , Glándulas Salivales/metabolismo , Transducción de Señal/fisiología , Células Madre/metabolismo , Agonistas Adrenérgicos beta/farmacología , Antagonistas Adrenérgicos beta/farmacología , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Humanos , Isoproterenol/farmacología , Organoides/citología , Organoides/metabolismo , Glándulas Salivales/citología , Salivación/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Células Madre/citologíaRESUMEN
BACKGROUND AND PURPOSE: Preclinical models are much needed to assess the effect of novel radio-sensitizers or mitigators on radiation dose limiting lung toxicity. Albeit showing radiation-induced lung pathologies, current mouse models lack the sensitivity to do so. Using micro image-guided radiotherapy (µIGRT) techniques, we aimed to establish murine models which enable the sensitive detection of lung damage aggravation and characterized functional, radiological and histological responses. MATERIALS AND METHODS: Right lungs of C57Bl/6J mice were irradiated using µIGRT with doses from 15 to 27â¯Gy and with 21â¯Gy and cisplatin as a radio-sensitizer in a second study. Mice were sacrificed for histological and pathological assessment at different time-points post-IR. Lung density was determined using the integrated micro cone-beam CT (µCBCT). Lung function was measured by double-chamber-plethysmography. RESULTS: µIGRT resulted in accurate deposition of the radiation dose in the right lung only as determined by É£H2AX staining. Lung fibrosis was confirmed by pathological assessments and increased significantly at 21â¯Gy as determined by automated quantification of histochemical analyses. Lung function was affected in a dose-dependent manner. µCBCT-determined lung densities increased significantly over time in the irradiated lungs and showed a strong radiation dose-dependence. Importantly, the µCBCT analyses allowed the detection of additional lung damage caused by 3â¯Gy dose increments or by the combination with cisplatin. CONCLUSION: µCBCT after right lung µIGRT enables the sensitive detection of effects inflicted by relative small dose increments or radio-sensitizers. Our preclinical model therefore facilitates the determination of lung damage exacerbation for the safety assessment of novel RT-drug combinations.
Asunto(s)
Tomografía Computarizada de Haz Cónico/métodos , Lesión Pulmonar/diagnóstico por imagen , Pulmón/diagnóstico por imagen , Pulmón/efectos de la radiación , Traumatismos por Radiación/diagnóstico por imagen , Animales , Modelos Animales de Enfermedad , Fraccionamiento de la Dosis de Radiación , Lesión Pulmonar/etiología , Masculino , Ratones , Ratones Endogámicos C57BL , Fibrosis PulmonarRESUMEN
PURPOSE: To study the ability of a prophylactic pilocarpine administration to preserve the rat parotid gland function after unilateral irradiation with graded doses of X-rays. METHODS: The right parotid gland of male albino Wistar rats was irradiated with single doses of X-rays (10-30 Gy, at 1.5 Gy min(-1)). Pilocarpine (4 mg/kg) was administered intraperitoneally, 1 hour prior to irradiation. Saliva samples of both left and right parotid gland were collected by means of miniaturized Lashley cups 4 days before and 3, 7, 10, and 30 days after irradiation. The parotid salivary flow rate (microl/min) was used as a parameter for the assessment of parotid gland function. RESULTS: Our data confirm that a single prophylactic treatment of pilocarpine can attenuate radiation-induced loss of gland function. Surprisingly, the effect of pilocarpine was not restricted to the irradiated gland only. Pilocarpine also enhanced the flow rate in the contralateral, nonirradiated gland. The latter effect was found for all doses above 10 Gy and became apparent around 7 days after the radiation treatment. The effectiveness of pilocarpine to attenuate function loss in the irradiated gland decreased with increasing dose and was lost after single doses of 30 Gy. CONCLUSIONS: Our data provide direct evidence that increasing the compensatory potential of the nondamaged gland, at least in part, underlies the "radioprotective effect" of pilocarpine in case of unilateral radiation. The ability of pilocarpine to ameliorate the early radiation-induced impairment of the parotid gland function in the irradiated gland may therefore be dependent on the remaining number of functional cells, and thus on the volume of the gland that lies within the radiation portal.
Asunto(s)
Agonistas Muscarínicos/farmacología , Glándula Parótida/efectos de los fármacos , Glándula Parótida/efectos de la radiación , Pilocarpina/farmacología , Traumatismos Experimentales por Radiación/prevención & control , Animales , Inyecciones Intraperitoneales , Masculino , Agonistas Muscarínicos/administración & dosificación , Glándula Parótida/fisiología , Pilocarpina/administración & dosificación , Traumatismos Experimentales por Radiación/etiología , Traumatismos Experimentales por Radiación/fisiopatología , Ratas , Ratas Wistar , Salivación/efectos de los fármacos , Salivación/efectos de la radiaciónRESUMEN
1. In the portal vein of the freely moving unanaesthetized rat, the existence of presynaptically located inhibitory muscarinic receptors was investigated by use of the muscarinic agonist methacholine (MCh) 2. Infusion of MCh (0.3 micrograms min-1) did not significantly inhibit the endogenous noradrenaline (NA) overflow in portal plasma. However, after inducing high intra-synaptic concentrations of NA by blocking the presynaptic alpha 2-adrenoceptors with yohimbine (1 mg kg-1), MCh (0.3 microgram min-1) was able to reduce the yohimbine-induced enhanced NA overflow by 38%. 3. The MCh-induced inhibition was almost completely abolished after blockade of the presynaptic muscarinic receptors with atropine (0.6 mg kg-1). 4. During electrical stimulation of the portal vein nervous plexus the evoked NA overflow was strongly inhibited (95%) during MCh-infusion (0.3 microgram min-1). Again atropine (0.6 mg kg-1) was able to reverse this inhibition. 5. These results show the existence of presynaptic muscarinic receptors inhibiting endogenous NA overflow from the portal vein nervous plexus under conditions of enhanced sympathetic activity in the freely moving rat.
Asunto(s)
Músculo Liso Vascular/metabolismo , Norepinefrina/metabolismo , Receptores Muscarínicos/fisiología , Sinapsis/metabolismo , Animales , Atropina/farmacología , Estimulación Eléctrica , Técnicas In Vitro , Masculino , Compuestos de Metacolina/farmacología , Músculo Liso Vascular/efectos de los fármacos , Vena Porta/efectos de los fármacos , Vena Porta/metabolismo , Ratas , Ratas Endogámicas , Receptores Muscarínicos/efectos de los fármacos , Sinapsis/efectos de los fármacos , Yohimbina/farmacologíaRESUMEN
1. The duration of the facilitatory effect of adrenaline on the electrically evoked overflow of noradrenaline was studied in the portal vein of permanently adreno-demedullated freely moving rats. 2. Rats were infused with adrenaline (20 or 100 ng min-1) for 2 h. After an interval of 1 h, when plasma adrenaline had returned to undetectable levels, electrical stimulation resulted in an enhanced catecholamine overflow amounting to 219% (noradrenaline) and 241% (noradrenaline plus adrenaline) of control (saline infusion), respectively. 3. When stimulation was applied again, in the same animal, at 24, 48 and 72 h after the first stimulation episode, the evoked noradrenaline overflow was 150, 111 and 102% (after 20 ng ml-1 adrenaline) and 158, 134 and 105% (after 100 ng min-1 adrenaline) of control. 4. The beta 2-adrenoceptor antagonist, ICI 118,551 (0.3 mg kg-1), blocked the facilitatory effect obtained after the 100 ng min-1 adrenaline infusion on all days. 5. The results show that adrenaline, after being taken up by and released from sympathetic nerve terminals, is able to facilitate the evoked noradrenaline overflow through activation of prejunctional beta 2-adrenoceptors for at least 48 h after administration.
Asunto(s)
Monoaminas Biogénicas/farmacología , Epinefrina/farmacología , Músculo Liso Vascular/inervación , Unión Neuromuscular/fisiología , Norepinefrina/fisiología , Vena Porta/inervación , Transmisión Sináptica/fisiología , Médula Suprarrenal/fisiología , Antagonistas Adrenérgicos beta/farmacología , Animales , Estimulación Eléctrica , Masculino , Músculo Liso Vascular/efectos de los fármacos , Unión Neuromuscular/efectos de los fármacos , Norepinefrina/farmacología , Vena Porta/efectos de los fármacos , Propanolaminas/farmacología , Ratas , Ratas Wistar , Transmisión Sináptica/efectos de los fármacosRESUMEN
1. In the portal vein of permanently cannulated, freely moving, unanaesthetized rats, methacholine (MCh) is able to inhibit the electrically-evoked endogenous noradrenaline (NA) overflow. This inhibition is mediated by presynaptic inhibitory muscarinic heteroreceptors. 2. By use of pirenzepine, 4-diphenylacetoxy-N-methylpiperidine methobromide (4-DAMP) and AF-DX 116 as M1-, M3-, and M2-selective antagonists respectively, the MCh (0.1 microM)-induced inhibition of the electrically-evoked NA overflow could be reversed to the control stimulation value dose-dependently. 3. The potency order of the antagonists was: 4-DAMP greater than AF-DX 116 greater than pirenzepine, pIC50 values being 8.50, 7.96 and 7.01, respectively. 4. From these results it was concluded that the inhibitory presynaptic heteroreceptors in the portal vein of conscious unrestrained rats are of the cardiac M2-subtype.
Asunto(s)
Músculo Liso Vascular/metabolismo , Norepinefrina/metabolismo , Receptores Muscarínicos/metabolismo , Animales , Catecolaminas/metabolismo , Cromatografía Líquida de Alta Presión , Relación Dosis-Respuesta a Droga , Electroquímica , Masculino , Músculo Liso Vascular/efectos de los fármacos , Piperidinas/farmacología , Pirenzepina/análogos & derivados , Pirenzepina/farmacología , Vena Porta/efectos de los fármacos , Vena Porta/metabolismo , Ratas , Ratas Endogámicas , Receptores Muscarínicos/efectos de los fármacos , Sinapsis/efectos de los fármacos , Sinapsis/metabolismoRESUMEN
It was studied whether differences in acute radiosensitivity exist between parotid and submandibular/sublingual glands. The results revealed that salivary flow rates decreased dramatically during the first 2 weeks of radiotherapy. Neither recovery nor significant differences were observed between the production of saliva from the parotid and submandibular/sublingual glands during the 13 weeks observation period.
Asunto(s)
Neoplasias de Cabeza y Cuello/radioterapia , Glándulas Salivales/efectos de la radiación , Salivación/efectos de la radiación , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Neoplasias de Cabeza y Cuello/complicaciones , Humanos , Masculino , Persona de Mediana Edad , Xerostomía/complicacionesRESUMEN
PURPOSE: The purpose of this study was to explore regional differences in radiosensitivity of rat lung using lung function and computed tomography (CT) density as endpoints. METHODS: At first, CT scans were used to determine rat lung volumes. The data obtained enabled the design of accurate collimators to irradiate 50% of the total lung volume for the apex, base, left, right, mediastinal and lateral part of the lung. Male Wistar rats were irradiated with a single dose of 18 Gy of orthovoltage X-rays. Further rat thorax CT scans were made before and 4, 16, 26, and 52 weeks after irradiation to measure in vivo lung density changes indicative of lung damage. To evaluate overall lung function, breathing frequencies were measured biweekly starting 1 week before irradiation. RESULTS: Qualitative analysis of the CT scans showed clear density changes for all irradiated lung volumes, with the most prominent changes present in the mediastinal and left group at 26 weeks after radiation. Quantitative analysis using average density changes of whole lungs did not adequately describe the differences in radiation response between the treated groups. However, analysis of the density changes of the irradiated and non-irradiated regions of interest (ROI) more closely matched with the qualitative observations. Breathing frequencies (BF) were only increased after 50% left lung irradiation, indicating that the hypersensitivity of the mediastinal part as assessed by CT analysis, does not result in functional changes. CONCLUSIONS: For both BF and CT (best described by ROI analysis), differences in regional lung radiosensitivity were observed. The presentation of lung damage either as function loss or density changes do not necessarily coincide, meaning that for each endpoint the regional sensitivity may be different.
Asunto(s)
Pulmón/efectos de la radiación , Absorciometría de Fotón , Animales , Pulmón/diagnóstico por imagen , Pulmón/fisiopatología , Masculino , Dosis de Radiación , Ratas , Ratas Wistar , Respiración/efectos de la radiación , Tomografía Computarizada por Rayos XRESUMEN
Radiotherapy of head and neck cancer frequently damages the salivary glands. Prophylactic administration of the muscarinic receptor agonist pilocarpine reduces subsequent radiation damage to the salivary glands in rats, but its effects on tumor cell radiosensitivity and tumor regrowth after irradiation had not been assessed. In the current study, we first tested the effect of pilocarpine on clonogenic cell survival in vitro. No effect of pilocarpine on radiosensitivity was observed in a panel of cell lines either with or without expression of muscarinic receptors. Second, a single dose of pilocarpine known to protect salivary gland tissue from radiation damage was given to rats transplanted with subcutaneously growing rhabdomyosarcomas 1 h prior to irradiation with a single dose of 35 Gy. No alterations in growth delay were detected (26 +/- 2 days for controls compared to 26 +/- 2 days for pilocarpine treatment). Our data indicate that pilocarpine pretreatment, which has been shown previously to protect salivary glands from radiation, does not protect tumor cells or tumors. Use of this drug therefore may lead to therapeutic gain in the treatment of head and neck cancer.
Asunto(s)
Carcinoma de Células Escamosas/radioterapia , División Celular/efectos de la radiación , Agonistas Muscarínicos/farmacología , Pilocarpina/farmacología , Rabdomiosarcoma/radioterapia , Neoplasias de las Glándulas Salivales/radioterapia , Glándulas Salivales/efectos de la radiación , Animales , Carcinoma de Células Escamosas/patología , División Celular/efectos de los fármacos , Relación Dosis-Respuesta en la Radiación , Humanos , Ratones , Traumatismos por Radiación/prevención & control , Ratas , Receptor Muscarínico M1 , Receptores Muscarínicos/genética , Receptores Muscarínicos/fisiología , Proteínas Recombinantes/metabolismo , Rabdomiosarcoma/patología , Neoplasias de las Glándulas Salivales/patología , Glándulas Salivales/efectos de los fármacos , Glándulas Salivales/patología , Factores de Tiempo , Transfección , Células Tumorales CultivadasRESUMEN
To investigate whether secretory granules play a role in the radiosensitivity of the salivary glands of rats, parotid acinar cells, submandibular acinar cells and/or submandibular granular convoluted tubule (GCT) cells were degranulated prior to irradiation. Degranulation of GCT cells was obtained by pretreatment with phenylephrine (5 mg/kg, t = -60 min) and methacholine (3.75 mg/kg, t = -120 min). Degranulation of acinar cells was attained by pretreatment with isoproterenol (5 mg/kg, t = -90 min). Combinations of pretreatments were also tested. Irradiation was performed with a single dose of 15 Gy of X rays. Samples of parotid and submandibular/sublingual saliva were collected 4 days prior to and 1, 3, 6, 10 and 30 days after irradiation. Pretreatment with phenylephrine, isoproterenol and methacholine plus phenylephrine resulted in less radiation damage to parotid gland function as indicated by the lag phase and flow rate. Since the pretreatment with phenylephrine and phenylephrine plus methacholine did not degranulate parotid gland acinar cells, the observed protective effect on this gland cannot be explained by the "degranulation concept." Furthermore, salivary gland function was significantly greater 3 days after irradiation as a result of pretreatment with phenylephrine and phenylephrine plus methacholine compared to rats given only radiation. This may indicate recovery from damage rather than a reduced amount of initial damage. The sparing was most obvious for the later effects (6-30 days). Submandibular/sublingual gland function was improved significantly after pretreatment with methacholine plus phenylephrine, although no increase in degranulation of GCT cells was observed compared to pretreatment with phenylephrine alone, again not favoring the degranulation concept. The results indicate that the secretory granules do not play the often-assumed important role in the radiosensitivity of the salivary gland. The mechanism underlying the observed improvement of salivary gland function may involve second messenger-induced increases in proliferation of salivary gland cells resulting in recovery of tissue after the irradiation.
Asunto(s)
Gránulos Citoplasmáticos/efectos de la radiación , Protección Radiológica , Saliva/metabolismo , Glándulas Salivales/fisiología , Glándulas Salivales/efectos de la radiación , Animales , Peso Corporal/efectos de la radiación , Gránulos Citoplasmáticos/efectos de los fármacos , Gránulos Citoplasmáticos/ultraestructura , Isoproterenol/farmacología , Masculino , Cloruro de Metacolina/farmacología , Glándula Parótida/fisiología , Glándula Parótida/efectos de la radiación , Fenilefrina/farmacología , Ratas , Ratas Wistar , Saliva/efectos de la radiación , Glándulas Salivales/efectos de los fármacos , Glándula Sublingual/fisiología , Glándula Sublingual/efectos de la radiación , Glándula Submandibular/fisiología , Glándula Submandibular/efectos de la radiación , Factores de Tiempo , Rayos XRESUMEN
Although the salivary glands have a low rate of cell turnover, they are relatively radiosensitive. To study the possible mechanism behind this inherent radiosensitivity, a rat model was developed in which saliva can be collected after local irradiation of the parotid gland without the use of anesthetics or stressful handling. Saliva secretion was induced by the partial muscarinic receptor agonist pilocarpine (0.03-3 mg/kg) with or without pretreatment with the beta-adrenoceptor antagonist propranolol (2.5 mg/kg), or the full muscarinic receptor agonist methacholine (0.16-16 mg/min), and measured during 5 min per drug dose before and 1, 3, 6 and 10 days after irradiation. The maximal secretory response induced by pilocarpine plus propranolol was increased compared to that with pilocarpine alone but did not reach the level of methacholine-induced secretion, which was about five times higher. One day after irradiation a decrease in maximal pilocarpine-induced secretion was observed (-22%) using the same dose of pilocarpine that induces 50% of the maximal response (ED(50)), in both the absence and presence of propranolol, indicating that the receptor-drug interaction was not affected by the radiation at this time. The secretory response to methacholine 1 day after irradiation, however, was normal. At day 3 after irradiation, the maximal methacholine-induced secretion was also affected, whereas pilocarpine (+/-propranolol)-induced maximal secretion decreased further. At day 6 after irradiation, maximal secretory responses had declined to approximately 50% regardless of the agonist used, whereas ED(50) values were still unaffected. No net acinar cell loss was observed within the first 10 days after irradiation, and this therefore could not account for the loss in function. The results indicate that radiation does not affect cell number or receptor-drug interaction, but rather signal transduction, which eventually leads to the impaired response. We hypothesize that the early radiation effect, within 3 days, may be membrane damage affecting the receptor-G-protein signaltransfer. Later critical damage, however, is probably of a different nature and may be located in the second-messenger signal transduction pathway downstream from the G protein, not necessarily involving cellular membranes.
Asunto(s)
Glándula Parótida/efectos de la radiación , Receptores Muscarínicos/fisiología , Antagonistas Adrenérgicos beta/farmacología , Animales , Peso Corporal/efectos de la radiación , Masculino , Cloruro de Metacolina/farmacología , Agonistas Muscarínicos/farmacología , Glándula Parótida/efectos de los fármacos , Glándula Parótida/metabolismo , Pilocarpina/farmacología , Propranolol/farmacología , Ratas , Ratas Wistar , Saliva/metabolismoRESUMEN
Previous studies on spontaneously hypertensive rats (SHR) have yielded inconsistent information about functional aberrations of the presynaptic alpha 2- and beta 2-adrenoceptor-mediated modulation of sympathetic neurotransmitter release. In the present investigation we studied the capacity of presynaptic beta 2-adrenoceptors that enhance noradrenaline (NA) release in the portal vein of freely moving, unanesthetized SHR and normotensive Wistar rats (WR) using the beta 2-selective agonist fenoterol. The results show that the presynaptic beta 2-adrenoceptor population in SHR responds to significantly lower dosages of fenoterol than that in WR. The reason for this enhanced action, however, could not be attributed to the beta 2-adrenoceptor itself, nor to a diminished neuronal uptake of NA, but to a diminished responsiveness of the presynaptic alpha 2-adrenoceptor. Stimulation of presynaptic alpha 2-adrenoceptors with oxymetazoline (45 micrograms/min) decreased basal NA levels by 46% in WR and by 3% in SHR. Blockade of alpha 2-adrenoceptors, using 0.5 mg/kg yohimbine, induced a 4.86-fold rise in the basal NA level in WR but only a 1.89-fold rise in SHR. A subsequent dose of fenoterol, however, resulted in a further 2.5- and 2.6-fold rise in WR and SHR, respectively, indicating that there is a normal presynaptic beta 2-adrenoceptor population in the vasculature of SHR.
Asunto(s)
Hipertensión/fisiopatología , Músculo Liso Vascular/efectos de los fármacos , Receptores Adrenérgicos alfa/efectos de los fármacos , Receptores Adrenérgicos beta/efectos de los fármacos , Animales , Cocaína/farmacología , Fenoterol/farmacología , Técnicas In Vitro , Masculino , Norepinefrina/sangre , Norepinefrina/metabolismo , Oximetazolina/farmacología , Vena Porta/efectos de los fármacos , Ratas , Ratas Endogámicas SHR , Ratas Endogámicas , Yohimbina/farmacologíaRESUMEN
The effect of adrenaline on the electrically evoked noradrenaline overflow in the portal vein of adrenal demedullated freely moving rats was studied. Adrenaline (100 ng/min) was infused for 2 h into the portal vein. After a 1-h interval when plasma adrenaline had returned to pre-infusion undetectable levels, the portal vein nervus plexus was electrically stimulated. During stimulation (2 Hz, 3 ms, 5 mA) adrenaline and noradrenaline were released. The stimulus-evoked noradrenaline overflow was facilitated to 194% of the control-evoked overflow (infusion of saline). Total catecholamine overflow (noradrenaline plus adrenaline) was enhanced to 258%. The facilitation of the evoked overflow of both noradrenaline and adrenaline was blocked by the selective beta 2-adrenoceptor antagonist, ICI 118,551 (0.3 mg/kg). Cocaine (2.5 mg/kg plus 0.05 mg/kg per min) infused together with adrenaline prevented the evoked release of adrenaline and no facilitation of the stimulus-induced noradrenaline overflow occurred. Inhibition of prejunctional inhibitory alpha 2-adrenoceptors with yohimbine (0.5 mg/kg) further enhanced, to 577%, the electrically evoked catecholamine overflow after adrenaline infusion. The results demonstrate that adrenaline can be taken up by sympathetic nerve endings through cocaine-sensitive uptake carriers, and is released from these nerves during electrical stimulation of the portal vein nervus plexus. Neuronally released adrenaline can strongly facilitate electrically evoked neurotransmitter overflow through activation of prejunctional beta 2-adrenoceptors.
Asunto(s)
Epinefrina/metabolismo , Vena Porta/inervación , Receptores Adrenérgicos beta/fisiología , Animales , Cocaína/farmacología , Estimulación Eléctrica , Epinefrina/farmacología , Hipertensión/etiología , Masculino , Ratas , Ratas Wistar , Yohimbina/farmacologíaRESUMEN
The effect of intravenously applied (-)-adrenaline, taken up by and released from sympathetic nerves, on swimming exercise-induced noradrenaline overflow in permanently cannulated adrenal demedullated rats was studied. Adrenaline (100 ng/min) was infused for 2 h, during which a plasma concentration of 500 pg/ml (approximately 2.5 nM) was reached. One hour later plasma adrenaline had returned to undetectable levels. During swimming, adrenaline was released into the plasma in concentrations up to 133 pg/ml and the noradrenaline concentration was markedly enhanced as well. The total catecholamine increase amounted to 178% of control (saline infusion) in the first 3 min of swimming and 165% for the whole 20 min. Cocaine (2.5 mg/kg plus 0.05 mg/kg/min), infused together with adrenaline and continued throughout the experiment, prevented the exercise-induced release of adrenaline and no increase in plasma noradrenaline concentration was observed. Yohimbine (0.25 mg/kg) strongly further enhanced the exercise-induced overflow of both noradrenaline and adrenaline. This further increase was completely blocked by the selective beta 2-adrenoceptor antagonist ICI 118,551 ((+/-)-1-[(2,3-dihydro-7-methyl-1H-inden-4-yl)oxy]-3-[(1-methyleth yl) amino]-2-butanol) (1.0 mg/kg). These results demonstrate that adrenaline can be taken up by sympathetic nerve endings through cocaine-sensitive uptake carriers and is released from these nerves during swimming exercise. Neuronally released adrenaline markedly enhances exercise-induced catecholamine overflow through activation of prejunctional beta 2-adrenoceptors.
Asunto(s)
Epinefrina/farmacología , Norepinefrina/metabolismo , Receptores Adrenérgicos beta 2/metabolismo , Antagonistas Adrenérgicos beta/administración & dosificación , Antagonistas Adrenérgicos beta/farmacología , Animales , Cocaína/administración & dosificación , Cocaína/farmacología , Sinergismo Farmacológico , Epinefrina/administración & dosificación , Epinefrina/sangre , Masculino , Norepinefrina/sangre , Condicionamiento Físico Animal , Propanolaminas/farmacología , Ratas , Ratas Wistar , Receptores Adrenérgicos beta 2/efectos de los fármacos , Natación/fisiología , Sistema Nervioso Simpático/efectos de los fármacos , Sistema Nervioso Simpático/metabolismo , Yohimbina/administración & dosificación , Yohimbina/farmacologíaRESUMEN
The effects of intraportal infusions of different doses of neuropeptide Y, its selective neuropeptide Y Y1 receptor analogue, [Leu31,Pro34]neuropeptide Y, and the Y2-selective C-terminal fragment, neuropeptide Y-(18-36), on basal and electrically evoked noradrenaline overflow in the portal vein as well as on mean arterial pressure and heart rate were investigated in permanently instrumented freely moving rats. Neuropeptide Y dose dependently (2-2000 ng/kg/min) attenuated the electrically evoked noradrenaline overflow and almost complete blockade was reached at the highest dose used. [Leu31,Pro34]Neuropeptide Y also dose dependently (20-20,000 ng/kg/min) attenuated the evoked overflow, reaching a maximum of 55% inhibition at the highest dose (20,000 ng/kg/min). Neuropeptide Y-(18-36) attenuated the evoked release only at 20,000 ng/kg/min (by 46%). Only at the highest dose did neuropeptide Y (2000 ng/kg/min) and [Leu31,Pro34]neuropeptide Y (20,000 ng/kg/min) significantly enhance mean arterial pressure and decrease heart rate and basal plasma noradrenaline levels, the latter two effects being due to the baroreceptor reflex. Neuropeptide Y-(18-36) did not influence these parameters at all doses used. The results indicate the presence of prejunctional neuropeptide Y Y1 receptors, and possibly the coexistence of Y1 and Y2 receptors, in the portal vein of freely moving rats, which in conjunction are able to inhibit markedly electrically evoked noradrenaline overflow. Postjunctional neuropeptide Y receptors mediating an increase in blood pressure in the freely moving rat are solely of the Y1 subtype.