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Vpliv različnih načinov dodajanja teriparatida na izražanje genov PPARG in FABP4 med adipogeno diferenciacijo mezenhimskih matičnih celic
ID Cafuta, Urška (Avtor), ID Ostanek, Barbara (Mentor) Več o mentorju... Povezava se odpre v novem oknu, ID Vrščaj, Lucija Ana (Komentor)

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Izvleček
Mezenhimske matične celice (MSC) so vrsta matičnih celic, ki so pomembne za razvoj in obnavljanje mišično-skeletnega sistema organizma. Delujejo kot prekurzorske celice in se lahko diferencirajo v adipocite, osteoblaste in hondrocite. V organizmu stalno potekata adipogeneza in osteogeneza, številni procesi in signalne poti, ki spodbujajo eno in zavirajo drugo, in obratno. V starosti pride do neravnovesja med procesoma, in sicer se zviša stopnja diferenciacije MSC v adipocite namesto v osteoblaste. To pa lahko vodi v nastanek bolezni, kot je osteoporoza, za katero je zaradi zmanjšane kostne mase in sprememb v mikroarhitekturi kosti značilna povečana lomljivost kosti. Večjo kostno gradnjo spodbuja osteoanabolna učinkovina teriparatid, ki smo ga proučevali v našem raziskovalnem delu. Namen magistrske naloge je bil ugotoviti razlike v izražanju genov PPARG in FABP4, ki sodelujeta v kaskadi procesov adipogene diferenciacije MSC. Uporabili smo 41 vzorcev MSC, ki so bili podvrženi adipogeni diferenciaciji in kontinuirano ali intermitentno izpostavljeni teriparatidu. Ta posnema delovanje parathormona, ki pri intermitentni aplikaciji spodbuja kostno tvorbo, pri kontinuirani aplikaciji pa spodbuja razgradnjo kostnine. Z uporabo različnih načinov aplikacij teriparatida smo želeli ugotoviti njihov vpliv na adipogeno diferenciacijo MSC. Najprej smo iz vzorcev MSC izolirali RNA in jo prepisali v cDNA. Nato smo z metodo kvantitativne verižne reakcije s polimerizacijo v realnem času izmerili izražanje adipogenih označevalcev. Uspešnost adipogene diferenciacije smo ugotavljali tudi s histokemičnim barvanjem adipocitov. Pri obeh tarčnih genih smo zaznali najvišje izražanje v vzorcih, ki so bili kontinuirano izpostavljeni teriparatidu 7 dni, kar je bilo tudi v skladu z rezultati barvanja adipocitov. Ti so pokazali, da je adipogeneza uspešno potekla le pri vzorcih, ki so bili teriparatidu izpostavljeni kontinuirano, in pri vzorcih, ki teriparatidu niso bili izpostavljeni. Pri izražanju gena PPARG nismo izmerili večjih razlik med intermitentno in kontinuirano aplikacijo teriparatida. Pri izražanju gena FABP4 pa smo opazili enak vzorec izražanja za vse načine aplikacije teriparatida.

Jezik:Slovenski jezik
Ključne besede:mezenhimske matične celice, adipogena diferenciacija, osteoporoza, teriparatid, kvantitativna verižna reakcija s polimerizacijo v realnem času.
Vrsta gradiva:Magistrsko delo/naloga
Organizacija:FFA - Fakulteta za farmacijo
Leto izida:2024
PID:20.500.12556/RUL-155911 Povezava se odpre v novem oknu
Datum objave v RUL:24.04.2024
Število ogledov:192
Število prenosov:38
Metapodatki:XML RDF-CHPDL DC-XML DC-RDF
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Sekundarni jezik

Jezik:Angleški jezik
Naslov:Effects of different teriparatide treatments on PPARG and FABP4 gene expression during adipogenic differentiation of mesenchymal stem cells
Izvleček:
Mesenchymal stem cells (MSCs) are a type of stem cells that are important for the development and regeneration of the body's musculoskeletal system. They act as precursor cells and can differentiate into adipocytes, osteoblasts, and chondrocytes. Adipogenesis and osteogenesis are constantly taking place in the body, many processes and signaling pathways that stimulate one and inhibit the other and vice versa. In old age, an imbalance between the two processes occurs, namely the rate of differentiation of MSCs into adipocytes instead of osteoblasts increases. This can lead to diseases such as osteoporosis, which is characterized by increased bone fragility due to reduced bone mass and changes in bone microarchitecture. Greater bone building is stimulated by the osteoanabolic agent teriparatide, which we studied in our research work. The aim of the master's thesis was to determine the differences in the expression of PPARG and FABP4 genes, which participate in the cascade of MSC adipogenic differentiation processes. We used 41 MSC samples that underwent adipogenic differentiation and were continuously or intermittently exposed to teriparatide. It mimics the action of parathormone, which stimulates bone formation with intermittent application and bone breakdown with continuous application. Using different methods of teriparatide application, we wanted to determine their influence on the adipogenic differentiation of MSCs. First, RNA was isolated from MSC samples and transcripted into cDNA. Then, we measured the expression of adipogenic markers using the quantitative real-time polymerization chain reaction method. The success of adipogenic differentiation was also determined by histochemical staining of adipocytes. For both target genes, we detected the highest expression in samples that were continuously exposed to teriparatide for 7 days, which was also in accordance with the results of adipocyte staining. They showed that adipogenesis was successful only in samples that were continuously exposed to teriparatide and in samples that were not exposed to teriparatide. We did not measure any major differences in the expression of the PPARG gene between intermittent and continuous administration of teriparatide. In the expression of the FABP4 gene, the same pattern of expression was observed for all methods of teriparatide application.

Ključne besede:mesenchymal stem cells, adipogenic differentiation, osteoporosis, teriparatide, quantitative realtime polymerase chain reaction.

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