Vpliv diferenciacije skeletnomišičnih celic na izražanje citokinov iz družine interlevkina-6

Spruk, Teja (author), Pirkmajer, Sergej (mentor)

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.

• Language: Slovenian
• Keywords: skeletna mišica, mioblasti, mišične cevčice, interlevkin-6
• Work type: Bachelor thesis/paper
• Typology: 2.11 - Undergraduate Thesis
• Organization: FKKT - Faculty of Chemistry and Chemical Technology
• Year: 2023
• PID: 20.500.12556/RUL-149553
• COBISS.SI-ID: 164425987
• Publication date in RUL: 07.09.2023

Secondary language

• Language: English
• Title: The effect of differentiation of skeletal muscle cells on expression of IL-6 family cytokines

Abstract

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.

• Keywords: skeletal muscle, myoblasts, myotubes, interleukin-6