Cytoskeletal Involvement in Hypothermic Renal Preservation Injury

Mangino, Martin J.; Tian, Tian; Ametani, Mary S.; Lindell, Susanne L.; Southard, James H.

doi:10.1097/tp.0b013e31815fed17

Cited by 19 publications

(29 citation statements)

References 63 publications

(21 reference statements)

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“…LDH release was determined with a CytoTox 96 assay kit to assess membrane integrity. Some cells that were plated in 6 well plates were extracted with a differential moesin extraction buffer after reperfusion to measure moesin disassociation as previously described in detail for ezrin [16]. …”

Section: Methodsmentioning

confidence: 99%

“…The functional significance of ERM proteins in hypothermic preservation injury is suggested by a series of studies of ezrin expression and functionality in renal tubular epithelium [16;24;25]. Ezrin disassociates from the actin cytoskeleton proportional to the degree of preservation injury in renal tubules [16].…”

Section: Introductionmentioning

confidence: 99%

“…Ezrin disassociates from the actin cytoskeleton proportional to the degree of preservation injury in renal tubules [16]. Ezrin is protective in preservation injury by virtue of its degree of expression and its binding configuration.…”

Section: Introductionmentioning

confidence: 99%

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Moesin functionality in hypothermic liver preservation injury

Tian¹,

Lindell²,

Kowalski³

et al. 2014

Cryobiology

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The objective of this study was to determine how expression and functionality of the cytoskeletal linker protein moesin is involved in hepatic hypothermic preservation injury. Mouse livers were cold stored in University of Wisconsin (UW) solution and reperfused on an Isolated Perfused Liver (IPL) device for one hour. Human hepatocytes (HepG2) and human or murine Sinusoidal Endothelial Cells (SECs) were cold stored and rewarmed to induce hypothermic preservation injury. The cells were transfected with: wild type moesin, an siRNA duplex specific for moesin, and the moesin mutants T558D and T558A. Tissue and cell moesin expression and its binding to actin were determined by western blot. Liver IPL functional outcomes deteriorated proportional to the length of cold storage, which correlated with moesin disassociation from the actin cytoskeleton. Cell viability (LDH and WST-8) in the cell models progressively declined with increasing preservation time, which also correlated with moesin disassociation. Transfection of a moesin containing plasmid or an siRNA duplex specific for moesin into HepG2 cells resulted in increased and decreased moesin expression, respectively. Overexpression of moesin protected while moesin knock-down potentiated preservation injury in the HepG2 cell model. Hepatocytes expressing the T558A (inactive) and T558D (active) moesin binding mutants demonstrated significantly more and less preservation injury, respectively. Cold storage time dependently caused hepatocyte detachment from the matrix and cell death, which was prevented by the T558D active moesin mutation. In conclusion, moesin is causally involved in hypothermic liver cell preservation injury through control of its active binding molecular functionality.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Moesin functionality in hypothermic liver preservation injury

Tian¹,

Lindell²,

Kowalski³

et al. 2014

Cryobiology

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“…The longer the cold storage is, the greater the changes on sodium-potassium adenosine triphosphate (Na/K-ATP) and in the proteins cytoskeleton, such as ezrin. All these events cause timedependent lipid membranes injury, disorganizing the nephron tubular function during the cold storage, contributing with the damage that occurs during rewarming 7 . Cold storage induces cell death due to the appearance of iron-dependent reactive oxygen species, with apoptosis in the rewarming period.…”

Section: Introductionmentioning

confidence: 99%

Oxidative stress evaluation of ischemia and reperfusion in kidneys under various degrees of hypothermia in rats

Santos

Koff

Filho³

et al. 2013

Acta Cir. Bras.

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PURPOSE:To design an animal model of ischemia-reperfusion (I/R) in kidneys and evaluate the role that predetermined ranges of local hypothermia plays on markers of stress-oxydative as well as on histologic sections. METHODS:Twenty eight male rats Wistar, under general anesthesia, undergone right nephrectomy (G0, control group) followed by left kidney ischemia during 40 min. Four temperatures groups were designed, with seven animals randomized for each group: normothermic (G1, ±37 o C), mild hypothermia (G2, 26 o C), moderate hypothermia (G3, 15 o C) and deep hypothermia (G4, 4 o C). Left kidney temperature was assessed with an intraparenchymal probe. Left nephrectomy was performed after 240 min of reperfusion. After I/R a blood sample was obtained for f2-IP. Half of each kidney was sent to pathological evaluation and half to analyze CAT, SOD, TBARS, NO 3 , NO 2 . RESULTS:Histopathology showed that all kidneys under I/R were significantly more injured than the G0 (p<0.001). TBARS had increased levels in all I/R groups compared with the G0 (p<0.001). CAT had a significant difference (p<0.03) between G1 and G4. Finally, no difference was found on SOD, NO 3 , NO 2 nor on f2-IP. CONCLUSION:This model of I/R was efficient to produce oxidative-stress in the kidney, showing that 4ºC offered significant decrease in free radicals production, although tissue protection was not observed.

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“…As the cell swells during ischemia, the swelling causes injury of the plasma and mitochondrial membranes and rupture (22;23), especially after having been weakened by concomitant disassembly of key cytoskeletal proteins during ischemia (20;21;24). As the parenchymal cells in the liver (mainly hepatocytes) swell, they compress the sinusoidal vascular spaces in aggregate, which decreases the diameter of the sinusoids and increases resistance to blood flow.…”

Section: Discussionmentioning

confidence: 99%

Donor gluconate rescues livers from uncontrolled donation after cardiac death

Limkemann

Lindell

Reichstetter

et al. 2016

Surgery

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Background Ischemia from organ preservation or donation causes cells and tissues to swell due to loss of energy-dependent mechanisms of control of cell volume. These volume changes cause substantial preservation injury, because preventing these changes by adding cell impermeants to preservation solutions decreases preservation injury. The objective of this study was to assess if this effect could be realized early in uncontrolled donation after cardiac death (DCD) livers by systemically loading donors with gluconate immediately after death to prevent accelerated swelling injury during the warm ischemia period prior to liver retrieval. Methods Uncontrolled DCD rat livers were cold-stored in UW solution for 24 hours and reperfused on an isolated perfused liver (IPL) device for two hours or transplanted into a rat as an allograft for 7 days. Donors were pre-treated with a solution of the impermeant gluconate or a saline control immediately after cardiac death. Livers were retrieved after 30 minutes. Results In-vivo, gluconate-infused in donors immediately before or after cardiac death prevented DCD induced increases in total tissue water (TTW), decreased vascular resistance, increased oxygen consumption and ATP synthesis, increased bile production, decreased LDH release, and decreased histology injury scores after reperfusion on the IPL relative to saline-treated DCD controls. In the transplant model, donor gluconate pretreatment significantly decreased both ALT the first day after transplantation and total bilirubin the seventh day after transplantation. Conclusions Cell and tissue swelling plays a key role in preservation injury of uncontrolled DCD livers, which can be mitigated by early administration of gluconate solutions to the donor immediately after death.

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Cytoskeletal Involvement in Hypothermic Renal Preservation Injury

Abstract: Hypothermic storage preservation causes disruption of key cytoskeletal elements in kidney tubules, which contributes causally to the injury at rewarming.

Cited by 19 publications

References 63 publications

Moesin functionality in hypothermic liver preservation injury

Moesin functionality in hypothermic liver preservation injury

Oxidative stress evaluation of ischemia and reperfusion in kidneys under various degrees of hypothermia in rats

Donor gluconate rescues livers from uncontrolled donation after cardiac death

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