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GSK3β and VDAC Involvement in ER Stress and Apoptosis Modulation during Orthotopic Liver Transplantation
Mohamed Amine Zaouali1,2,3, Arnau Panisello4, Alexandre Lopez5
1Experimental Hepatic Ischemia-Reperfusion Unit, Institut d'Investigacions Biomèdiques de Barcelona (IIBB), Spanish National Research Council (CSIC), Barcelona 08036, Catalonia, Spain. daminzaouali12@yahoo.fr.
Abstract:
We investigated the involvement of glycogen synthase kinase-3β (GSK3β) and the voltage-dependent anion channel (VDAC) in livers subjected to cold ischemia-reperfusion injury (I/R) associated with orthotopic liver transplantation (OLT). Rat livers were preserved in University of Wisconsin (UW) and Institute Georges Lopez (IGL-1) solution, the latter enriched or not with trimetazidine, and then subjected to OLT. Transaminase (ALT) and HMGB1 protein levels, glutamate dehydrogenase (GLDH), and oxidative stress (MDA) were measured. The AKT protein kinase and its direct substrates, GSK3β and VDAC, as well as caspases 3, 9, and cytochrome C and reticulum endoplasmic stress-related proteins (GRP78, pPERK, ATF4, and CHOP), were determined by Western blot. IGL-1+TMZ significantly reduced liver injury. We also observed a significant phosphorylation of AKT, which in turn induced the phosphorylation and inhibition of GSK3β. In addition, TMZ protected the mitochondria since, in comparison with IGL-1 alone, we found reductions in VDAC phosphorylation, apoptosis, and GLDH release. All these results were correlated with decreased ER stress. Addition of TMZ to IGL-1 solution increased the tolerance of the liver graft to I/R injury through inhibition of GSK3β and VDAC, contributing to ER stress reduction and cell death prevention.
Insights
Trimetazidine (TMZ) added to IGL-1 solution significantly reduced liver injury during cold ischemia-reperfusion (I/R) in orthotopic liver transplantation (OLT). This protection involved inhibiting glycogen synthase kinase-3β (GSK3β) and voltage-dependent anion channel (VDAC), reducing ER stress and preventing cell death.
Area of Science:
- Hepatology
- Transplantation Immunology
- Cellular Stress Response
Background:
- Cold ischemia-reperfusion injury (I/R) is a major challenge in orthotopic liver transplantation (OLT), leading to significant graft damage.
- Glycogen synthase kinase-3β (GSK3β) and voltage-dependent anion channel (VDAC) are implicated in cellular injury pathways.
- Understanding the molecular mechanisms of I/R injury is crucial for developing protective strategies.
Purpose of the Study:
- To investigate the role of GSK3β and VDAC in liver I/R injury during OLT.
- To evaluate the protective effects of trimetazidine (TMZ) in liver preservation solutions against I/R injury.
- To elucidate the molecular mechanisms underlying TMZ's protective effects.
Main Methods:
- Rat livers were preserved using University of Wisconsin (UW) and Institute Georges Lopez (IGL-1) solutions, with IGL-1 optionally enriched with TMZ.
- Livers underwent orthotopic liver transplantation (OLT) followed by cold ischemia-reperfusion (I/R).
- Biochemical markers (ALT, GLDH, MDA), protein levels (HMGB1, AKT, GSK3β, VDAC, caspases, cytochrome C, ER stress proteins), and phosphorylation states were assessed via Western blot.
Main Results:
- IGL-1 solution enriched with TMZ significantly reduced liver injury markers (ALT, GLDH, MDA) compared to IGL-1 alone.
- TMZ treatment led to increased AKT phosphorylation, subsequently inhibiting GSK3β phosphorylation.
- TMZ reduced VDAC phosphorylation, apoptosis, and GLDH release, correlating with decreased endoplasmic reticulum (ER) stress markers (GRP78, pPERK, ATF4, CHOP).
Conclusions:
- Trimetazidine (TMZ) enhances liver graft tolerance to I/R injury in OLT by inhibiting GSK3β and VDAC.
- TMZ's protective mechanism involves reducing ER stress and preventing apoptosis.
- This study highlights TMZ as a promising agent for improving liver preservation strategies in transplantation.
