Branched-chain amino acids and mitochondrial bioenergetics in C2C12 myoblasts

Johdanto. Haaraketjuiset aminohapot (HKAH, leusiini, isoleusiini, valiini) ja niiden aineenvaihdunta on liitetty parempaan aineenvaihdunnalliseen terveyteen, pidempään elinkaareen, parantuneeseen glukoosi- ja rasva-aineenvaihduntaan sekä mitokondrioiden biogeneesiin. Toisaalta kohonnut HKAH:n pitois...

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Main Author: Lehtonen, Elias
Other Authors: Liikuntatieteellinen tiedekunta, Faculty of Sport and Health Sciences, Liikunta- ja terveystieteet, Sport and Health Sciences, Jyväskylän yliopisto, University of Jyväskylä
Format: Master's thesis
Language:eng
Published: 2019
Subjects:
Online Access: https://jyx.jyu.fi/handle/123456789/64109
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author Lehtonen, Elias
author2 Liikuntatieteellinen tiedekunta Faculty of Sport and Health Sciences Liikunta- ja terveystieteet Sport and Health Sciences Jyväskylän yliopisto University of Jyväskylä
author_facet Lehtonen, Elias Liikuntatieteellinen tiedekunta Faculty of Sport and Health Sciences Liikunta- ja terveystieteet Sport and Health Sciences Jyväskylän yliopisto University of Jyväskylä Lehtonen, Elias Liikuntatieteellinen tiedekunta Faculty of Sport and Health Sciences Liikunta- ja terveystieteet Sport and Health Sciences Jyväskylän yliopisto University of Jyväskylä
author_sort Lehtonen, Elias
datasource_str_mv jyx
description Johdanto. Haaraketjuiset aminohapot (HKAH, leusiini, isoleusiini, valiini) ja niiden aineenvaihdunta on liitetty parempaan aineenvaihdunnalliseen terveyteen, pidempään elinkaareen, parantuneeseen glukoosi- ja rasva-aineenvaihduntaan sekä mitokondrioiden biogeneesiin. Toisaalta kohonnut HKAH:n pitoisuus veressä on liitetty lihavuuteen, aineenvaihdunnallisiin sairauksiin, insuliiniresistenssiin sekä tyypin 2 diabetekseen. On ehdotettu, että muutokset HKAH hajotuksessa, ei niiden saannissa, on yhteydessä aineenvaihdunnallisiin sairauksiin. Tutkimuksen tavoitteena on tarkastella HKAH altistuksen vaikutuksia raajalihassolun mitokondrioiden bioenergetiikkaan C2C12 myoblasteissa. Menetelmät. C2C12 myoblastit altistettiin kasvaneelle HKAH pitoisuudelle (5 mM leusiini, 5 mM isoleusiini, 5mM valiini) 24 tunnin ajan, 4 tunnin ajan tai akuutisti ja niiden soluhengitystä verrattiin kontrolliryhmään korkean resoluution respirometrialla. Soluhengitystä tutkittiin sekä hiilihydraatti- että rasvapohjaisen aerobisen aineenvaihdunnan yhteydessä. Hiilihydraattipohjaisessa soluhengityksessä määritettiin fysiologinen (Routine), protonivuoto (Leak), Kompleksi I, Kompleksi I + II (OXPHOS), elektroninsiirtoketjun maksimikapasiteetti (ETS) sekä Kompleksi II linkittynyt soluhengitys. Rasvapohjaisessa hengityksessä määritettiin fysiologinen (Routine), protonivuoto (Leak), rasva-happojen oksidaatio (FAO) sekä ETS hengitysvaiheet. Lisäksi mitattiin proteiinikonsentraatio ja sitraattisyntaasin aktiivisuus. Hengitysarvot suhteutettiin solumäärään, proteiinikonsentraatioon sekä sitraattisyntaasin aktiivisuuteen. Lisäksi hengitysarvot suhteutettiin maksimaaliseen ETS kapasiteettiin. Tulokset. 24 tunnin HKAH altistus kasvatti tilastollisesti merkitsevästi ETS hengitystä hiilihydraattiaineenvaihdunnan osalta verrattuna kontrolliryhmään (p < 0.05), 4 tunnin ja akuutin altistuksen osalta ei havaittu eroja suhteessa kontrolliryhmään. 24 tunnin ryhmällä oli hiilihydraattiaineenvaihdunnan osalta tilastollisesti merkitsevästi matalampi OXPHOS hengitys (p < 0.01) suhteutettuna ETS hengitykseen verrattuna kontrolliryhmään. Rasva-aineenvaihdunnan osalta ei havaittu eroja hengitysvaiheissa eri ryhmien välillä. Sitraattisyntaasin aktiivisuudessa sekä proteiinikonsentraatioissa ei ollut eroja koeryhmien osalta. Johtopäätökset. HKAH altistus kasvattaa maksimaalista soluhengitystä hiilihydraatti-, mutta ei rasvahappopohjaisessa soluhengityksessä. HKAH vaikutus on yhteydessä altistusaikaan siten, että pitkä (24h) altistus kasvatti maksimaalista soluhengitystä, mutta lyhyt (4h) tai akuutti altistus eivät vaikuttaneet soluhengitykseen. HKAH voi olla mahdollisia hyötyjä hiilihydraattipohjaisessa soluhengityksessä. Sen sijaan negatiivisia vaikutuksia HKAH altistukseen ei havaittu soluhengityksen suhteen. Introduction. Branched-chain amino acids (BCAAs, leucine, isoleucine, valine) and their metabolism have been linked to improved metabolic health, prolonged lifespan, improved glucose and lipid metabolism and mitochondrial biogenesis. On the other hand, circulating BCAAs and their metabolites are connected to obesity, metabolic disease, insulin resistance and type 2 diabetes mellitus. It has been suggested that alterations in BCAA catabolism rather than intake are connected to metabolic disease. This study aims to establish the effect of increased exposure to BCAAs on skeletal muscle mitochondrial bioenergetics in C2C12 myoblasts. Methods. C2C12 myoblasts were exposed to increased BCAA concentration (5 mM Leucine, 5 mM Isoleucine, 5 mM Valine) for 24 hours, 4 hours or acutely, compared to a control group. Cellular respiration was then measured with high-resolution respirometry in order to establish the effect of BCAAs on mitochondrial bioenergetics. Cellular respiration was measured via both carbohydrate and fatty acid dependent pathways in separate experiments. In carbohydrate dependent respiration, Routine, Leak, Complex I, Complex I + II (OXPHOS), electron transport system capacity (ETS) and uncoupled Complex II respiratory states were determined. In fatty acid dependent respiration, Routine, Leak, Fatty acid oxidation (FAO) and ETS respiratory states were defined. Additionally, protein concentration and citrate synthase activity of the samples were determined. Respiration values were normalized to cell count, protein concentration and citrate synthase activity. Additionally, coupling control ratios were examined by normalizing respiratory values at different states to maximal ETS capacity. Results. 24 hour exposure to increased BCAA concentration increased maximal ETS respiratory state of carbohydrate dependent respiration statistically significantly compared to the control group (p < 0.05), 4 hour and acutely increased BCAA exposure did not affect cellular respiration in comparison to the control group. 24 h group had statistically significantly lower OXPHOS (p < 0.01) coupling control ratio compared to the control group. No differences were observed between the experimental groups in fatty acid based respiratory states. Citrate synthase activity and protein concentration did not differ in terms of the experimental groups. Conclusions. Increased exposure to BCAAs increases maximal cellular respiration in carbohydrate, but not fatty acid based oxidation. The effects are time-dependent in that prolonged (24h), but not short (4h) or acute exposure increased ETS respiratory state. BCAAs may improve aerobic metabolism of carbohydrates, no negative metabolic outcomes were observed.
first_indexed 2019-09-20T09:13:25Z
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Haaraketjuiset aminohapot (HKAH, leusiini, isoleusiini, valiini) ja niiden aineenvaihdunta on liitetty parempaan aineenvaihdunnalliseen terveyteen, pidemp\u00e4\u00e4n elinkaareen, parantuneeseen glukoosi- ja rasva-aineenvaihduntaan sek\u00e4 mitokondrioiden biogeneesiin. Toisaalta kohonnut HKAH:n pitoisuus veress\u00e4 on liitetty lihavuuteen, aineenvaihdunnallisiin sairauksiin, insuliiniresistenssiin sek\u00e4 tyypin 2 diabetekseen. On ehdotettu, ett\u00e4 muutokset HKAH hajotuksessa, ei niiden saannissa, on yhteydess\u00e4 aineenvaihdunnallisiin sairauksiin. Tutkimuksen tavoitteena on tarkastella HKAH altistuksen vaikutuksia raajalihassolun mitokondrioiden bioenergetiikkaan C2C12 myoblasteissa. Menetelm\u00e4t. C2C12 myoblastit altistettiin kasvaneelle HKAH pitoisuudelle (5 mM leusiini, 5 mM isoleusiini, 5mM valiini) 24 tunnin ajan, 4 tunnin ajan tai akuutisti ja niiden soluhengityst\u00e4 verrattiin kontrolliryhm\u00e4\u00e4n korkean resoluution respirometrialla. Soluhengityst\u00e4 tutkittiin sek\u00e4 hiilihydraatti- ett\u00e4 rasvapohjaisen aerobisen aineenvaihdunnan yhteydess\u00e4. Hiilihydraattipohjaisessa soluhengityksess\u00e4 m\u00e4\u00e4ritettiin fysiologinen (Routine), protonivuoto (Leak), Kompleksi I, Kompleksi I + II (OXPHOS), elektroninsiirtoketjun maksimikapasiteetti (ETS) sek\u00e4 Kompleksi II linkittynyt soluhengitys. Rasvapohjaisessa hengityksess\u00e4 m\u00e4\u00e4ritettiin fysiologinen (Routine), protonivuoto (Leak), rasva-happojen oksidaatio (FAO) sek\u00e4 ETS hengitysvaiheet. Lis\u00e4ksi mitattiin proteiinikonsentraatio ja sitraattisyntaasin aktiivisuus. Hengitysarvot suhteutettiin solum\u00e4\u00e4r\u00e4\u00e4n, proteiinikonsentraatioon sek\u00e4 sitraattisyntaasin aktiivisuuteen. Lis\u00e4ksi hengitysarvot suhteutettiin maksimaaliseen ETS kapasiteettiin. \nTulokset. 24 tunnin HKAH altistus kasvatti tilastollisesti merkitsev\u00e4sti ETS hengityst\u00e4 hiilihydraattiaineenvaihdunnan osalta verrattuna kontrolliryhm\u00e4\u00e4n (p < 0.05), 4 tunnin ja akuutin altistuksen osalta ei havaittu eroja suhteessa kontrolliryhm\u00e4\u00e4n. 24 tunnin ryhm\u00e4ll\u00e4 oli hiilihydraattiaineenvaihdunnan osalta tilastollisesti merkitsev\u00e4sti matalampi OXPHOS hengitys (p < 0.01) suhteutettuna ETS hengitykseen verrattuna kontrolliryhm\u00e4\u00e4n. Rasva-aineenvaihdunnan osalta ei havaittu eroja hengitysvaiheissa eri ryhmien v\u00e4lill\u00e4. Sitraattisyntaasin aktiivisuudessa sek\u00e4 proteiinikonsentraatioissa ei ollut eroja koeryhmien osalta. Johtop\u00e4\u00e4t\u00f6kset. HKAH altistus kasvattaa maksimaalista soluhengityst\u00e4 hiilihydraatti-, mutta ei rasvahappopohjaisessa soluhengityksess\u00e4. HKAH vaikutus on yhteydess\u00e4 altistusaikaan siten, ett\u00e4 pitk\u00e4 (24h) altistus kasvatti maksimaalista soluhengityst\u00e4, mutta lyhyt (4h) tai akuutti altistus eiv\u00e4t vaikuttaneet soluhengitykseen. HKAH voi olla mahdollisia hy\u00f6tyj\u00e4 hiilihydraattipohjaisessa soluhengityksess\u00e4. Sen sijaan negatiivisia vaikutuksia HKAH altistukseen ei havaittu soluhengityksen suhteen.", "language": "fi", "element": "description", "qualifier": "abstract", "schema": "dc"}, {"key": "dc.description.abstract", "value": "Introduction. Branched-chain amino acids (BCAAs, leucine, isoleucine, valine) and their metabolism have been linked to improved metabolic health, prolonged lifespan, improved glucose and lipid metabolism and mitochondrial biogenesis. On the other hand, circulating BCAAs and their metabolites are connected to obesity, metabolic disease, insulin resistance and type 2 diabetes mellitus. 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spellingShingle Lehtonen, Elias Branched-chain amino acids and mitochondrial bioenergetics in C2C12 myoblasts branched-chain amino acids oxidative phosphorylation high-resolution respirometry. metabolic disease Liikuntafysiologia Exercise Physiology 5011 mitokondriot aineenvaihdunta rasvahapot aminohapot insuliiniresistenssi hengitys lihakset glukoosi lipidit mitochondria metabolism fatty acids amino acids insulin resistance respiration muscles glucose lipids
title Branched-chain amino acids and mitochondrial bioenergetics in C2C12 myoblasts
title_full Branched-chain amino acids and mitochondrial bioenergetics in C2C12 myoblasts
title_fullStr Branched-chain amino acids and mitochondrial bioenergetics in C2C12 myoblasts Branched-chain amino acids and mitochondrial bioenergetics in C2C12 myoblasts
title_full_unstemmed Branched-chain amino acids and mitochondrial bioenergetics in C2C12 myoblasts Branched-chain amino acids and mitochondrial bioenergetics in C2C12 myoblasts
title_short Branched-chain amino acids and mitochondrial bioenergetics in C2C12 myoblasts
title_sort branched chain amino acids and mitochondrial bioenergetics in c2c12 myoblasts
title_txtP Branched-chain amino acids and mitochondrial bioenergetics in C2C12 myoblasts
topic branched-chain amino acids oxidative phosphorylation high-resolution respirometry. metabolic disease Liikuntafysiologia Exercise Physiology 5011 mitokondriot aineenvaihdunta rasvahapot aminohapot insuliiniresistenssi hengitys lihakset glukoosi lipidit mitochondria metabolism fatty acids amino acids insulin resistance respiration muscles glucose lipids
topic_facet 5011 Exercise Physiology Liikuntafysiologia aineenvaihdunta amino acids aminohapot branched-chain amino acids fatty acids glucose glukoosi hengitys high-resolution respirometry. metabolic disease insuliiniresistenssi insulin resistance lihakset lipidit lipids metabolism mitochondria mitokondriot muscles oxidative phosphorylation rasvahapot respiration
url https://jyx.jyu.fi/handle/123456789/64109 http://www.urn.fi/URN:NBN:fi:jyu-201905222711
work_keys_str_mv AT lehtonenelias branchedchainaminoacidsandmitochondrialbioenergeticsinc2c12myoblasts