20.500.12556/RUL-149553 Vpliv diferenciacije skeletnomišičnih celic na izražanje citokinov iz družine interlevkina-6 The effect of differentiation of skeletal muscle cells on expression of IL-6 family cytokines Skupna značilnost IL-6 in ostalih predstavnikov družine citokinov IL-6 je, da kot del receptorskega kompleksa uporabljajo glikoprotein (gp130). Ta je lahko v receptorskem kompleksu prisoten v obliki homodimera ali heterodimera. IL-6 in IL-11 za signalizacijo uporabljata homodimer gp130, ostali člani pa heterodimere gp130 v kombinaciji z enim od ostalih β-receptorjev (LIFRβ ali OSMRβ). V okviru našega laboratorijskega dela smo skeletnomišične kulture diferencirali do različnih stopenj in v teh prekinili celične procese ter jih zamrznili. S pomočjo kvantitativne verižne reakcije s polimerazo v realnem času smo nato preverili izražanje sedmih predstavnikov iz družine citokinov IL-6 – IL-6, LIF, OSM in CTF-1, CNTF, CLCF-1 in CRLF-1. Pri tem smo hoteli ugotoviti, kateri od teh se sploh izražajo v primarnih človeških skeletnomišičnih celicah in kako se to izražanje spreminja glede na stopnjo diferenciacije. Pred tem smo preverili izražanje znanega diferenciacijskega markerja MYH2 in tako pokazali, da so se celice resnično diferencirale. Z našimi rezultati smo pokazali, da se IL-6, LIF, CTF-1, CNTF, CLCF-1 in CRLF-1 izražajo v skeletnomišičnih celicah, pri čemer se je izražanje IL-6, LIF, CLCF-1 in CRLF-1 zmanjšalo z diferenciacijo, izražanje CNTF se je povečalo, raven izražanja CTF-1 pa se je najprej povečala, do konca diferenciacije pa spet zmanjšala. OSM se je izražal v vseh diferenciacijskih stopnjah, vendar je raven njegovega izražanja tako nizka, da sprememb kvantitativno ni bilo mogoče ovrednotiti. Interleukin-6 and other members of the IL-6 cytokine family are defined by their common use of the glycoprotein 130 (gp130). It can be a component of the receptor complex in the form of a homodimer or a heterodimer. IL-6 and IL-11 use a gp130 homodimer for signalization and the other cytokines from this family use a heterodimer, composed of one gp130 subunit and one of the other β-receptors (LIFRβ or OSMRβ). In the scope of our laboratory work, we differentiated skeletal muscle cultures to different degrees and checked the expression of seven members of the IL-6 cytokine family – IL-6, LIF, OSM in CTF-1, CNTF, CLCF-1 in CRLF-1. With that, we investigated which of these were expressed in human skeletal muscle cells and how their expression changed in relation to the degree of differentiation. We also checked the expression of a known differentiation marker MYH2 and showed that the cells really did differentiate. We showed that IL-6, LIF, CTF-1, CNTF, CLCF-1 in CRLF-1 are expressed in skeletal muscle cells, whereby the expression of IL-6, LIF, CLCF-1 and CRLF-1 decreased or tended to decrease with differentiation, Converesely, the expression of CNTF increased, while the expression of CTF-1 first increased and then decreased by the end of differentiaton. Although OSM was expressed in all differentiation stages, its levels were too low for reliable quantification. skeletna mišica mioblasti mišične cevčice interlevkin-6 skeletal muscle myoblasts myotubes interleukin-6 true false false Slovenski jezik Angleški jezik Diplomsko delo/naloga 2023-09-07 11:27:07 2023-09-07 11:27:07 2023-09-14 11:37:03 0000-00-00 00:00:00 2023 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 22850 164425987 Spruk_Teja_-_Vpliv_diferenciacije_skeletnomisicnih_celic_na_izrazanje_citokinov_iz_druzine_IL-6.pdf Spruk_Teja_-_Vpliv_diferenciacije_skeletnomisicnih_celic_na_izrazanje_citokinov_iz_druzine_IL-6.pdf 1 786FB82EDF8D2D0B59BC955C62B14E5C 8aaa7b330152ba52be82486436693f0c00af0d6d09393cf33fc14323f1a947a0 b5b13e47-4d60-11ee-9934-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=174037 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0