HDL-mediated cholesterol efflux and macrophage polarization in human THP-1 macrophages

HDL:t (eng. high-density lipoproteins, HDLs) ovat monimuotoinen joukko lipoproteiineja, jotka välittävät useita alempaan sydän- ja verisuonitautiriskiin yhteydessä olevia toimintoja. HDL:n pääasiallinen tehtävä on kuljettaa kolesterolia perifeerisistä soluista maksaan eritettäväksi käänteisessä kole...

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Päätekijä: Karttunen, Elina
Muut tekijät: Matemaattis-luonnontieteellinen tiedekunta, Faculty of Sciences, Bio- ja ympäristötieteiden laitos, Department of Biological and Environmental Science, Jyväskylän yliopisto, University of Jyväskylä
Aineistotyyppi: Pro gradu
Kieli:eng
Julkaistu: 2024
Aiheet:
Linkit: https://jyx.jyu.fi/handle/123456789/95991
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author Karttunen, Elina
author2 Matemaattis-luonnontieteellinen tiedekunta Faculty of Sciences Bio- ja ympäristötieteiden laitos Department of Biological and Environmental Science Jyväskylän yliopisto University of Jyväskylä
author_facet Karttunen, Elina Matemaattis-luonnontieteellinen tiedekunta Faculty of Sciences Bio- ja ympäristötieteiden laitos Department of Biological and Environmental Science Jyväskylän yliopisto University of Jyväskylä Karttunen, Elina Matemaattis-luonnontieteellinen tiedekunta Faculty of Sciences Bio- ja ympäristötieteiden laitos Department of Biological and Environmental Science Jyväskylän yliopisto University of Jyväskylä
author_sort Karttunen, Elina
datasource_str_mv jyx
description HDL:t (eng. high-density lipoproteins, HDLs) ovat monimuotoinen joukko lipoproteiineja, jotka välittävät useita alempaan sydän- ja verisuonitautiriskiin yhteydessä olevia toimintoja. HDL:n pääasiallinen tehtävä on kuljettaa kolesterolia perifeerisistä soluista maksaan eritettäväksi käänteisessä kolesterolin kuljetuksessa. Prosessin ensimmäinen vaihe on kolesterolin ulosvirtaus, jossa kolesteroli kulkeutuu soluista HDL-partikkeleille. Liikunnan on todettu lisäävän HDL-partikkelien määrää ja tehostavan kolesterolin ulosvirtausta, mutta aerobisen kuntotason vaikutus kolesterolin ulosvirtaukseen on vielä epäselvä. Makrofagit ovat joukko immuunisoluja, jotka kykenevät muuttamaan fenotyyppiään vasteena ympäristöönsä polarisaatioksi kutsutussa prosessissa. Valtimonkovettumataudissa tulehdusta edistävien M1-makrofagien määrä verisuonten seinämissä lisääntyy. HDL:lla on tulehdusta ehkäiseviä ominaisuuksia ja sen on hiirillä todettu lisäävän tulehdusta estävien M2-tyypin makrofagien ilmentymistä. HDL:n rooli ihmisen makrofagien polarisaatiossa on vielä epäselvä. Tämän työn tavoitteena oli tutkia HDL:n toiminnallisuutta kolesterolin ulosvirtauksen ja makrofagien polarisaation kautta ihmisen THP-1 makrofageissa. Ihmisen THP-1 makrofagit käsiteltiin hyvä- ja huonokuntoisten koehenkilöiden HDL:lla. Kolesterolin ulosvirtauskapasiteetti mitattiin fluoresoivasti leimattua kolesterolia hyödyntävän menetelmän avulla. Makrofagien polarisaatiota ja fenotyyppejä HDL-käsittelyiden jälkeen tutkittiin qPCR:n sekä immunoleimauksen ja konfokaalimikroskopian avulla. Kolesterolin ulosvirtaus oli tehokkaampaa matalan aerobisen kuntotason ryhmässä. HDL ei suoraan vaikuttanut makrofagien polarisaatioon kohti M2 fenotyyppiä, mutta vähensi VEGF-markkerin ilmentymistä viitaten HDL:n mahdolliseen rooliin tulehduksellisen angiogeneesin ehkäisemisessä. Tulokset viittaavat siihen, että aerobisella kuntotasolla ja kolesterolin ulosvirtauskapasiteetilla on yhteys, mutta yhteyden taustamekanismit vaativat lisätutkimuksia. HDL:n vaikutus ihmisen makrofagien polarisaatioon jää vielä epäselväksi ja vaatii lisätutkimuksia. Lisäksi makrofagien polarisaatioprotokolla vaatii vielä lisätutkimuksia ja kehitystä. High-density lipoproteins (HDLs) are a diverse group of lipoproteins with beneficial functions related to lower cardiovascular risk. They function to deliver cholesterol from peripheral tissues to liver for excretion in a process called reverse cholesterol transport. Cholesterol efflux is the first step of this process where cholesterol moves from the cell towards HDL. Exercise increases HDL levels and enhances cholesterol efflux, but the effect of aerobic fitness level on cholesterol efflux is less studied. Macrophages are plastic immune cells capable of phenotypic switching between inflammatory M1 and anti-inflammatory M2 phenotypes in a process called polarization. Macrophages play central roles in the pathogenesis of atherosclerosis. Atherosclerotic lesions are characterized by an imbalance of the M1 and M2 phenotypes, with M1 macrophages being more abundant. HDL increases the expression of M2 macrophages in mice but its effect on human macrophage polarization is less clear. The aim of this thesis was to study the functionality of HDL by examining the effect of aerobic fitness on HDL-mediated cholesterol efflux and the effect of HDL on macrophage polarization in human THP-1 macrophages. THP-1 macrophages were incubated with HDL from individuals of high and low aerobic fitness followed by measuring the cholesterol efflux capacity (CEC) with a fluorescent-labeled cholesterol kit. The expression of macrophage markers after HDL treatments was studied with qPCR and immunofluorescence. CEC was higher in the low aerobic fitness group. HDL did not directly modulate the polarization of macrophages towards the M2 phenotype but decreased the expression of VEGF in macrophages, indicating a possible role in the inhibition of inflammatory angiogenesis. In conclusion, the results indicate a difference in efflux capacity in relation to aerobic fitness level but the mechanisms behind the interaction require further investigation. The role of HDL in the polarization of human macrophages remains unclear and requires more research. Additionally, the macrophage polarization protocol requires further investigation and standardization.
first_indexed 2024-06-18T20:01:04Z
format Pro gradu
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HDL:n p\u00e4\u00e4asiallinen teht\u00e4v\u00e4 on kuljettaa kolesterolia perifeerisist\u00e4 soluista maksaan eritett\u00e4v\u00e4ksi k\u00e4\u00e4nteisess\u00e4 kolesterolin kuljetuksessa. Prosessin ensimm\u00e4inen vaihe on kolesterolin ulosvirtaus, jossa kolesteroli kulkeutuu soluista HDL-partikkeleille. Liikunnan on todettu lis\u00e4\u00e4v\u00e4n HDL-partikkelien m\u00e4\u00e4r\u00e4\u00e4 ja tehostavan kolesterolin ulosvirtausta, mutta aerobisen kuntotason vaikutus kolesterolin ulosvirtaukseen on viel\u00e4 ep\u00e4selv\u00e4. Makrofagit ovat joukko immuunisoluja, jotka kykenev\u00e4t muuttamaan fenotyyppi\u00e4\u00e4n vasteena ymp\u00e4rist\u00f6\u00f6ns\u00e4 polarisaatioksi kutsutussa prosessissa. Valtimonkovettumataudissa tulehdusta edist\u00e4vien M1-makrofagien m\u00e4\u00e4r\u00e4 verisuonten sein\u00e4miss\u00e4 lis\u00e4\u00e4ntyy. HDL:lla on tulehdusta ehk\u00e4isevi\u00e4 ominaisuuksia ja sen on hiirill\u00e4 todettu lis\u00e4\u00e4v\u00e4n tulehdusta est\u00e4vien M2-tyypin makrofagien ilmentymist\u00e4. HDL:n rooli ihmisen makrofagien polarisaatiossa on viel\u00e4 ep\u00e4selv\u00e4. T\u00e4m\u00e4n ty\u00f6n tavoitteena oli tutkia HDL:n toiminnallisuutta kolesterolin ulosvirtauksen ja makrofagien polarisaation kautta ihmisen THP-1 makrofageissa. Ihmisen THP-1 makrofagit k\u00e4siteltiin hyv\u00e4- ja huonokuntoisten koehenkil\u00f6iden HDL:lla. Kolesterolin ulosvirtauskapasiteetti mitattiin fluoresoivasti leimattua kolesterolia hy\u00f6dynt\u00e4v\u00e4n menetelm\u00e4n avulla. Makrofagien polarisaatiota ja fenotyyppej\u00e4 HDL-k\u00e4sittelyiden j\u00e4lkeen tutkittiin qPCR:n sek\u00e4 immunoleimauksen ja konfokaalimikroskopian avulla. Kolesterolin ulosvirtaus oli tehokkaampaa matalan aerobisen kuntotason ryhm\u00e4ss\u00e4. 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main_date_str 2024
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spellingShingle Karttunen, Elina HDL-mediated cholesterol efflux and macrophage polarization in human THP-1 macrophages aerobic fitness apolipoprotein A-1 atherosclerosis cholesterol gene expression high-density lipoprotein polarization reverse cholesterol transport M1 macrophages M2 macrophages Solu- ja molekyylibiologia Cell and molecular biology 4013 kolesteroli ateroskleroosi makrofagit HDL-kolesteroli lipoproteiinit apolipoproteiinit macrophages HDL cholesterol lipoproteins apolipoproteins
title HDL-mediated cholesterol efflux and macrophage polarization in human THP-1 macrophages
title_full HDL-mediated cholesterol efflux and macrophage polarization in human THP-1 macrophages
title_fullStr HDL-mediated cholesterol efflux and macrophage polarization in human THP-1 macrophages HDL-mediated cholesterol efflux and macrophage polarization in human THP-1 macrophages
title_full_unstemmed HDL-mediated cholesterol efflux and macrophage polarization in human THP-1 macrophages HDL-mediated cholesterol efflux and macrophage polarization in human THP-1 macrophages
title_short HDL-mediated cholesterol efflux and macrophage polarization in human THP-1 macrophages
title_sort hdl mediated cholesterol efflux and macrophage polarization in human thp 1 macrophages
title_txtP HDL-mediated cholesterol efflux and macrophage polarization in human THP-1 macrophages
topic aerobic fitness apolipoprotein A-1 atherosclerosis cholesterol gene expression high-density lipoprotein polarization reverse cholesterol transport M1 macrophages M2 macrophages Solu- ja molekyylibiologia Cell and molecular biology 4013 kolesteroli ateroskleroosi makrofagit HDL-kolesteroli lipoproteiinit apolipoproteiinit macrophages HDL cholesterol lipoproteins apolipoproteins
topic_facet 4013 Cell and molecular biology HDL cholesterol HDL-kolesteroli M1 macrophages M2 macrophages Solu- ja molekyylibiologia aerobic fitness apolipoproteiinit apolipoprotein A-1 apolipoproteins ateroskleroosi atherosclerosis cholesterol gene expression high-density lipoprotein kolesteroli lipoproteiinit lipoproteins macrophages makrofagit polarization reverse cholesterol transport
url https://jyx.jyu.fi/handle/123456789/95991 http://www.urn.fi/URN:NBN:fi:jyu-202406184757
work_keys_str_mv AT karttunenelina hdlmediatedcholesteroleffluxandmacrophagepolarizationinhumanthp1macrophages