Cytokine induced airway hyperreactivity characterized in an in-vitro model of asthma
(2005) In Lund University Faculty of Medicine Doctoral Dissertation Series- Abstract
- Asthma is a chronic disorder of the lower airways, characterized by inflammation and hyperreactivity. Pro-inflammatory cytokines like tumour necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) are known to play important roles in the pathogenesis, but our knowledge how these cytokines contribute to the development of airway hyperreactivity is limited.
The aims of the present study were to investigate relationships between TNF-alpha and IL-1beta, and airway smooth muscle contractility, focusing on the mechanism behind the development of airway hyperreactivity. To that end an in-vitro model for assessment of long-term effects of inflammatory mediators on the airway smooth muscle was developed. The model was... (More) - Asthma is a chronic disorder of the lower airways, characterized by inflammation and hyperreactivity. Pro-inflammatory cytokines like tumour necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) are known to play important roles in the pathogenesis, but our knowledge how these cytokines contribute to the development of airway hyperreactivity is limited.
The aims of the present study were to investigate relationships between TNF-alpha and IL-1beta, and airway smooth muscle contractility, focusing on the mechanism behind the development of airway hyperreactivity. To that end an in-vitro model for assessment of long-term effects of inflammatory mediators on the airway smooth muscle was developed. The model was based on the organ culture of murine tracheal segments in combination with subsequent evaluation in a myograph system, along with mRNA quantification and histology. Tissues were shown to maintain their contractile phenotype and morphological organization during a culture period up to 16 days.
Presence of TNF-alpha and IL-1beta, during 1, 2, 4 or 8 days of culture, increased the contractile response induced by 5-hydroxytryptamine (5-HT), des-Arg9-bradykinin and bradykinin in a time- and concentration-dependent manner, whereas the contractile response to sarafotoxin 6c (S6c) was decreased by the long-term presence of IL-1beta. Pharmacological analysis revealed that contractions induced by 5-HT, des-Arg9-bradykinin and bradykinin were mediated by 5HT2A, and bradykinin B1 and B2 receptors, respectively. Real-time PCR showed that TNF-alpha and IL-1beta induced an up-regulation of the bradykinin B1 and B2 receptors. In addition, IL-1beta caused a down-regulation of the endothelin B receptor (ETB).
Experiments with actinomycin D, a general transcriptional inhibitor and dexamethasone, a NF-kappaB / AP-1 inhibitor, along with analysis of the receptor mRNA expression revealed that the up-regulation of bradykinin B1 and B2 receptors was dependent on intact transcriptional mechanisms, whereas the increased 5-HT2A contraction appeared unaffected by transcriptional interference. 5-HT2A receptor-mediated contractions induced by IL-1beta involved the JNK and p38 mitogen-activated protein kinase (MAPK) pathways.
Experiments with the JNK inhibitor SP600125, the ERK 1/2 inhibitor PD98059 and the p38 inhibitor SB203580 suggested that JNK was a key molecule of vital importance for both TNF-alpha and IL-1beta induced bradykinin B1 and B2 receptor up-regulation, whereas both the ERK1/2 and JNK pathways were required for the IL-1beta induced down-regulation of the ETB receptors.
Remicade(registered), an anti-TNF-alpha antibody, inhibited IL-1beta-induced increases of the airway contractions, via inhibition of TNF-alpha and subsequent synthesis of the new receptors.
The present findings that pro-inflammatory cytokines, like TNF-alpha and IL-1beta, can induce airway hyperreactivity via interference with the intracellular MAPK signal transduction pathways might contribute to new concepts for the treatment of asthma. (Less) - Abstract (Swedish)
- Popular Abstract in Swedish
Astma är en sjukdom som karakteriseras av upprepade anfall av andningssvårigheter, orsakade av kramp, slemhinnesvullnad och segt sekret i luftvägarna. Den finns beskriven i texter redan från det antika Grekland och rapporteras idag vara en sjukdom med utbredning över hela vårt klot. Astma är den vanligaste kroniska sjukdomen bland barn och är oavsett ålder den vanligaste lungsjukdomen i världen.
Sjukdomen kännetecknas av en ökad känslighet sk. hyperreaktivitet för olika sorters stimuli, såväl specifika som pollen och ospecifika som tobaksrök och damm. Även under perioder när astmatikern känner sig bra, finns en ständigt pågående inflammation i luftrörens slemhinnor. Mycket... (More) - Popular Abstract in Swedish
Astma är en sjukdom som karakteriseras av upprepade anfall av andningssvårigheter, orsakade av kramp, slemhinnesvullnad och segt sekret i luftvägarna. Den finns beskriven i texter redan från det antika Grekland och rapporteras idag vara en sjukdom med utbredning över hela vårt klot. Astma är den vanligaste kroniska sjukdomen bland barn och är oavsett ålder den vanligaste lungsjukdomen i världen.
Sjukdomen kännetecknas av en ökad känslighet sk. hyperreaktivitet för olika sorters stimuli, såväl specifika som pollen och ospecifika som tobaksrök och damm. Även under perioder när astmatikern känner sig bra, finns en ständigt pågående inflammation i luftrörens slemhinnor. Mycket tyder på att det är denna inflammation som orsakar hyperreaktiviteten. Riktigt hur inflammationen uppstår och bibehålles är ännu inte klarlagt, men av hundratals olika signalsubstanser är inblandade och mycket forskning ägnas idag åt att kartlägga deras effekter och interaktioner.
I blod och lungsköljvätska från astmatiska patienter har man uppmätt stegrade nivåer av inflammatoriska signalsubstanser som TNF-alpha och IL-1beta. Man finner också förhöjda nivåer av bronksammandragande substanser som serotonin, bradykinin och endothelin-1. Därvid skapar möjlighet för interaktioner mellan inflammatoriska och bronksammandragande substanser, något som tros vara av betydelse för uppkomst och utveckling av hyperreaktivitet. Hur en eventuell sådan samverkan går till har dock varit ofullständigt känt.
Målsättning
Föreliggande avhandling syftar till att närmare kartlägga sambandet mellan inflammatoriska mediatorer och bronksammandragande substanser, med fokus på utveckling av luftvägshyperreaktivitet. Vi har härvid valt att titta närmare på TNF-alpha och IL-1beta, och deras relation till de serotonin, bradykinin, endothelin-1. Arbetet rör sig huvudsakligen kring tre frågeställningar:
1. Kan långvarig närvaro av TNF-alpha och IL-1beta påverka de luftvägssammandragande effekterna av serotonin, bradykinin och endothelin?
2. Vilka receptorer är inblandade i ett eventuellt förändrat sammandragnings-mönster?
3. Är det möjligt att närmare kartlägga och analysera de intracellulära signalvägar som kan tänkas vara inblandade i ett sådant skeende?
Resultat
För att svara på dessa frågor, började vi med att utveckla en astmamodell i mus där långtidseffekter av olika inflammatoriska mediatorer på luftvägarnas sammandragande förmåga kunde studeras. Ringsegment från musens luftstrupe isolerades och odlades under 1 till 8 dagar med eller utan närvaro av inflammatoriska mediatorer. Därefter monterades segmenten i vävnadsbad och effekterna av olika bronksammandragande substanser testades.
1. Man kunde härvid konstatera att närvaro av TNF-alpha eller IL-1beta påtagligt förstärkte den luftvägssammandragande effekten av serotonin och bradykinin, medan effekten av endothelin försvagades.
2. Genom farmakologisk analys kunde visas att denna förändring förmedlades via en ökad aktivitet av 5-HT2A, bradykinin B1 och B2 receptorer och en minskad endothelin B receptorers aktivitet. Aktivitetsförändringen var delvis knuten till förändringar i genuttrycket av receptorer i luftvägsmuskulaturen.
3. Det var vidare möjligt att i detalj kartlägga vilka intracellulära signalvägar som var inblandade och bland dessa intog den sk MAP kinas-vägen en central roll.
Kunskap om effekten av inflammatoriska mediatorer och deras signalvägar i luftvägarnas glatta muskulatur kan skapa de förutsättningar som krävs och nya möjligheter att minska och behandla hyperreaktivitet i luftvägarna hos astmapatienter. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/545890
- author
- Zhang, Yaping LU
- supervisor
- opponent
-
- Professor Simonsen, Ulf, Department of Pharmacology, University of Aarhus, Denmark
- organization
- publishing date
- 2005
- type
- Thesis
- publication status
- published
- subject
- keywords
- toxicology, Farmakologi, farmakognosi, MAPK, TNF-alpha, 5-HT, Respiratory system, Andningsorganen, Pharmacological sciences, pharmacognosy, pharmacy, interleukin-1beta, hyperresponsiveness, endothelin-1, airway, bradykinin, toxikologi, farmaci
- in
- Lund University Faculty of Medicine Doctoral Dissertation Series
- pages
- 165 pages
- publisher
- Laboratory of Clinical and Experimental Allergy Research, Department of Otorhinolaryngology, Institute of Clinical Science, Malmö University Hospital, Lund University, Sweden
- defense location
- Lilla Aulan, Entrance 59, Medicinskt Forskningscentrum, Malmö University Hospital, Malmö, Swden
- defense date
- 2005-12-15 13:00:00
- ISSN
- 1652-8220
- ISBN
- 91-85481-20-3
- language
- English
- LU publication?
- yes
- additional info
- Mikael Adner, Andrew C. Rose, Yaping Zhang, Karl Swärd, Mikael Benson, Rolf Uddman, Nigel P. Shankley and Lars-Olaf Cardell. 2002. An assay to evaluate the long-term effects of inflammatory mediators on murine airway smooth muscle: evidence that TNF-alpha up-regulates 5-HT2A-mediated contraction. British Journal of Pharmacology, vol 137 pp 971-982. Nature Publishing GroupYaping Zhang, Lars-Olaf Cardell and Mikael Adner. . IL-1beta induces murine airway hyper-responsiveness, via a non-transcriptional up-regulation of 5-HT2A mediated activity. (submitted)Yaping Zhang, Mikael Adner and Lars-Olaf Cardell. 2004. IL-1beta attenuates ETB receptor-mediated airway contractions in a murine in-vitro model of asthma: roles of endothelin converting enzyme and mitogen-activated protein kinase pathways. Clinical and Experimental Allergy, vol 34 pp 1480-1487. Blackwell Publishing Ltd.Yaping Zhang, Mikael Adner and Lars-Olaf Cardell. 2004. Up-regulation of bradykinin receptors in a murine in-vitro model of chronic airway inflammation European Journal of Pharmacology, vol 489 pp 117-126. Elsevier B.V.Yaping Zhang, Mikael Adner and Lars-Olaf Cardell. 2005. Glucocorticoids suppress transcriptional up-regulation of bradykinin receptors in a murine in-vitro model of chronic airway inflammation. Clinical and Experimental Allergy, vol 35 pp 531-538. Blackwell Publishing Ltd.Yaping Zhang, Mikael Adner and Lars-Olaf Cardell. . IL-1beta induced transcriptional up-regulation of bradykinin B1 and B2 receptors in murine airways. (submitted)
- id
- 3a265036-ff44-4e39-9a22-ec64d1ed51aa (old id 545890)
- date added to LUP
- 2016-04-01 15:47:10
- date last changed
- 2019-05-21 21:48:08
@phdthesis{3a265036-ff44-4e39-9a22-ec64d1ed51aa, abstract = {{Asthma is a chronic disorder of the lower airways, characterized by inflammation and hyperreactivity. Pro-inflammatory cytokines like tumour necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) are known to play important roles in the pathogenesis, but our knowledge how these cytokines contribute to the development of airway hyperreactivity is limited.<br/><br> <br/><br> The aims of the present study were to investigate relationships between TNF-alpha and IL-1beta, and airway smooth muscle contractility, focusing on the mechanism behind the development of airway hyperreactivity. To that end an in-vitro model for assessment of long-term effects of inflammatory mediators on the airway smooth muscle was developed. The model was based on the organ culture of murine tracheal segments in combination with subsequent evaluation in a myograph system, along with mRNA quantification and histology. Tissues were shown to maintain their contractile phenotype and morphological organization during a culture period up to 16 days.<br/><br> <br/><br> Presence of TNF-alpha and IL-1beta, during 1, 2, 4 or 8 days of culture, increased the contractile response induced by 5-hydroxytryptamine (5-HT), des-Arg9-bradykinin and bradykinin in a time- and concentration-dependent manner, whereas the contractile response to sarafotoxin 6c (S6c) was decreased by the long-term presence of IL-1beta. Pharmacological analysis revealed that contractions induced by 5-HT, des-Arg9-bradykinin and bradykinin were mediated by 5HT2A, and bradykinin B1 and B2 receptors, respectively. Real-time PCR showed that TNF-alpha and IL-1beta induced an up-regulation of the bradykinin B1 and B2 receptors. In addition, IL-1beta caused a down-regulation of the endothelin B receptor (ETB).<br/><br> <br/><br> Experiments with actinomycin D, a general transcriptional inhibitor and dexamethasone, a NF-kappaB / AP-1 inhibitor, along with analysis of the receptor mRNA expression revealed that the up-regulation of bradykinin B1 and B2 receptors was dependent on intact transcriptional mechanisms, whereas the increased 5-HT2A contraction appeared unaffected by transcriptional interference. 5-HT2A receptor-mediated contractions induced by IL-1beta involved the JNK and p38 mitogen-activated protein kinase (MAPK) pathways.<br/><br> <br/><br> Experiments with the JNK inhibitor SP600125, the ERK 1/2 inhibitor PD98059 and the p38 inhibitor SB203580 suggested that JNK was a key molecule of vital importance for both TNF-alpha and IL-1beta induced bradykinin B1 and B2 receptor up-regulation, whereas both the ERK1/2 and JNK pathways were required for the IL-1beta induced down-regulation of the ETB receptors.<br/><br> <br/><br> Remicade(registered), an anti-TNF-alpha antibody, inhibited IL-1beta-induced increases of the airway contractions, via inhibition of TNF-alpha and subsequent synthesis of the new receptors.<br/><br> <br/><br> The present findings that pro-inflammatory cytokines, like TNF-alpha and IL-1beta, can induce airway hyperreactivity via interference with the intracellular MAPK signal transduction pathways might contribute to new concepts for the treatment of asthma.}}, author = {{Zhang, Yaping}}, isbn = {{91-85481-20-3}}, issn = {{1652-8220}}, keywords = {{toxicology; Farmakologi; farmakognosi; MAPK; TNF-alpha; 5-HT; Respiratory system; Andningsorganen; Pharmacological sciences; pharmacognosy; pharmacy; interleukin-1beta; hyperresponsiveness; endothelin-1; airway; bradykinin; toxikologi; farmaci}}, language = {{eng}}, publisher = {{Laboratory of Clinical and Experimental Allergy Research, Department of Otorhinolaryngology, Institute of Clinical Science, Malmö University Hospital, Lund University, Sweden}}, school = {{Lund University}}, series = {{Lund University Faculty of Medicine Doctoral Dissertation Series}}, title = {{Cytokine induced airway hyperreactivity characterized in an in-vitro model of asthma}}, year = {{2005}}, }