| Abstract | Cilj istraživanja: Istražiti postoji li razlika u izražaju gena Lama 1, Tnc, Cldn1, Col2a1, Egfr, Per1, Per2, Per3, Reverbα, Spon2, Tef, Arntl, Mup7 i Nfil3 u mokraćnom mjehuru GF miševa u usporedbi s SPF miševima.
Animalni modeli i metode: Eksperimentalno istraživanje provedeno je u Laboratoriju za istraživanje raka Medicinskog fakulteta u Splitu. Analizirani su mokraćni mjehuri GF (N=12) i SPF (N=12) miševa soja C57BL/6, starih 6 tjedana. Bilo je 12 mužjaka i 12 ženki (polovica mužjaka je bila SPF, a polovica GF statusa, isto vrijedi i za ženke). Iz uzoraka je izolirana RNA, nakon čega je prevedena u cDNA koja je služila kao predložak za qRT-PCR. Dizajnirane su početnice za gene od interesa. Kao standard za genski izražaj korišten je gen za aktin.
Rezultati: U GF miševa značajno je povećan izražaj gena Per1 (p<0,0005), Tef (p<0,0005), Per2 (p<0,005) i Per3 (p<0,05) u usporedbi sa SPF miševima. U GF miševa značajno je smanjen izražaj gena Nfil3 (p<0,0005), Arntl (p<0,0005) i Spon2 (p<0,005) u odnosu na SPF miševe. U GF mužjaka statistički je značajno povećan izražaj gena Mup7 (p<0,05) u odnosu na SPF mužjake, dok ne postoji statistički značajna razlika između ženki. Izražaj gena Lama1, Tnc, Cldn1, Col2α1, Egfr i Reverbα nije promijenjenu GF miševa u odnosu na SPF miševe.
Zaključak: Mikrobiota mijenja izražaj nekih gena u mokraćnom mjehuru miševa. U odsustvu mikrobiote (u GF miševa) promijenio se izražaj većine gena koji kontroliraju cirkadijani ritam, dok je od svih testiranih gena izvanstanične tvari samo jedan bio promijenjen. Samo kod mužjaka se značajno promijenio izražaj gena Mup7, koji je dio feromonskog sustava miša. Buduće studije trebaju istražiti značaj opisanih promjena. |
| Abstract (english) | Objectives: To determine differences in expression of Lama 1, TNC, CLDN1, Col2a1, EGFR, Per1, Per2, Per3, Reverbα, Spon2, Tef, Arntl, Mup7 and Nfil3 genes between bladder tissue of germ-free and specific-pathogen free mice.
Animal models and methods: Experimental research was conducted at Laboratory for cancer research at University of Split School of Medicine. Bladder tissue of 6 weeks old GF (N=12) and SPF (N=12) wild type C57BL/6 mice was analised. There were 6 females and 6 males (half od males was GF and half was SPF, the same is true for females). RNA was isolated from tissue samples and then converted into cDNA by reverse transcription. cDNA was used as a template for qRT-PCR reaction. Primers were designed for genes of interest. Action gene was used as a housekeeping control.
Results: In GF mice gene expression was significantly increased for Per1 (p<0.0005), Tef (p<0.0005), Per2 (p<0.005) and Per3 (p<0.05) genes compared to SPF mice. Expression of Mup7gene was significantly increased in GF males (p<0.05) compared to SPF males. There was no significant difference in Mup7 expression between females. In GF females and males gene expression was significantly decreased for Nfil3 (p<0.0005), Arntl (p<0.0005) and Spon2 (p<0.005) genes compared to SPF females and males. Expression of genes Lama1, Tnc, Cldn1, Col2α1, Egfr and Reverbα was not changed in GF mice compared to SPF mice.
Conclusion: Microbiota is influencing gene expression in mice urinary bladder. Expression of most of the tested circadian rhytm genes is significantly changed in GF mice. Expression of only one (Spon2) extracellular matrix gene is significantly changed. Expression of Mup7 gene is increased in GF males. Future studies need to address relevance of presented findings. |
