Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Jakubauskiene, L.
Uterus Preservation: Reduction of Ischemia-Reperfusion Injury and Static Cold Storage Damage using Relaxin and Erythropoietin in Experimental Rat Uterus model
Doktoratsstudium der Medizinischen Wissenschaft; Humanmedizin; [ Dissertation ] Medizinische Universität Graz; 2023. pp. 101
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- Autor*innen der Med Uni Graz:
- Betreuer*innen:
- Leber Bettina
- Rosenkranz Alexander
- Stiegler Philipp
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- Abstract:
- Uterus transplantation (UTx) is the only treatment method for women with absolute uterine factor infertility and it enables women to be both genetical mothers and to experience pregnancy and childbearing. However, UTx is negatively affected by uterus static cold storage (SCS) and ischemia-reperfusion injury (IRI). Relaxin (RLX) and erythropoietin (EPO) are the substances that have already shown anti-inflammatory, anti-apoptotic and anti-fibrotic effects in several SCS and IRI models. This cumulative thesis aims to highlight the protective effect of RLX or/and EPO in rat uterus SCS and IRI models and to examine morphological and biochemical changes in rat uterine tissue. Additionally, we aimed to investigate gene panels, associated with oxidative stress, apoptosis and possible acting mechanisms of the experimental substances. Firstly, uterus SCS was investigated. Therefore, uterus horns retrieved from 15 Sprague Dawley rats were randomly assigned into three groups (n = 10/group) and kept at 4◦C in HTK-N solution without or with different additives: 10 IU/mL EPO or 20 nM RLX. Tissue samples were taken after 8 and 24 h of preservation. Secondly, uterus IRI was simulated. Therefore, eighty rats were randomly assigned into eight groups (n = 10/group) and pretreated with either saline, 5 μg/kg human relaxin-2, 4000 IU/kg recombinant human erythropoietin or their combination. Ischemia was achieved by clamping the aorta and ovarian arteries for 60 min, following 120 min of reperfusion and tissue sampling. For both parts uterine tissue histology, biochemical and immunohistochemical markers were analyzed. Additionally, gene panels were examined for tissues exposed to IRI. Uterine tissue morphology, MDA, SOD levels and the TUNEL-positive cell number showed severe damage in HTK-N solution without additives after 24 h of preservation during SCS. This damage was significantly attenuated by adding RLX to the preservation solution whereas EPO showed no favorable effect. Pretreatment with RLX preserved normal tissue morphology, reduced MDA levels, MPO- and TUNEL-positive cell count, preserved SOD activity and upregulated NICD and HES1 gene expression when compared to the control group during IRI. Pretreatment with EPO reduced MDA levels. In conclusion, our study shows that RLX as an additive to HTK-N solution can serve as an effective uterine tissue preservative in a SCS setting by inhibiting inflammation, apoptosis and preserving tissue morphology. Pretreatment with RLX, EPO or a combination of both EPO and RLX during IRI significantly alleviates uterine tissue damage. The combination of RLX with EPO was as effective as RLX alone, except the combination yielded a beneficial effect on anti-inflammatory gene expression.