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Cell injury after ischemia and reperfusion in the porcine kidney evaluated by radiolabelled microspheres, sestamibi, and lactadherin.

Pedersen SS, Keller AK, Nielsen MK, Jespersen B, Falborg L, Rasmussen JT, Heegaard CW, Rehling M - EJNMMI Res (2013)

Bottom Line: In WI240, the uptake of microspheres was severely reduced in both groups (17% ± 11% and 27% ± 17%).GFR was severely reduced in the post ischemic kidney in both groups.In both groups, the uptake of lactadherin was reduced (41% ± 8%, 17% ± 13%) but not different from the uptake of (153)Gd microspheres.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Clinical Medicine, Aarhus University, Aarhus, Denmark ; Department of Clinical Physiology and Nuclear Medicine, Aarhus University Hospital, Skejby, Denmark.

ABSTRACT

Background: The purpose of the present study was to quantify renal cell injury after ischemia and reperfusion in a pig model using (99m)Tc-lactadherin as a marker of apoptosis and (99m)Tc-sestamibi as a marker of mitochondrial dysfunction.

Methods: Thirty-four pigs were randomized into unilateral renal warm ischemia of 120 (WI120) or 240 min (WI240). The glomerular filtration rate (GFR) was calculated by renal clearance of (51)Cr-ethylenediaminetetraacetic acid, and apoptosis was quantified by immunohistochemical detection of caspase-3. After 240 min of reperfusion, intravenous (99m)Tc-lactadherin or (99m)Tc-sestamibi was injected simultaneously with (153)Gd microspheres into the aorta. Ex-vivo static planar images of the kidneys were acquired for determination of the differential renal function of tracer distribution using a gamma camera.

Results: In WI120, there was no significant difference in the uptake of microspheres in the ischemic and contralateral normal kidney indicating adequate perfusion (uptake in ischemic kidney relative to the sum of uptake in both kidneys; 46% ± 12% and 51% ± 5%). In WI240, the uptake of microspheres was severely reduced in both groups (17% ± 11% and 27% ± 17%). GFR was severely reduced in the post ischemic kidney in both groups. In both groups, the uptake of lactadherin was reduced (41% ± 8%, 17% ± 13%) but not different from the uptake of (153)Gd microspheres. Caspase-3-positive cell profiles were increased in the post-ischemic kidneys (p < 0.001) and increased as the length of ischemia increased (p = 0.003). In both WI120 and WI240, the amount of (99m)Tc-sestamibi in the ischemic kidney was significantly lower than the amount of (153)Gd microspheres (40 ± 5 versus 51 ± 5 and 20 ± 11 versus 27 ± 17; p < 0.05).

Conclusions: In an established pig model with unilateral renal warm ischemia, we found significantly reduced (99m)Tc-sestamibi uptake relative to perfusion in the kidneys exposed to ischemia indicating a potential ability to detect renal ischemic and reperfusion injuries. However, apoptosis was not detected using (99m)Tc-lactadherin in the post-ischemic kidneys despite increased number of caspase-3-positive cell profiles.

Trial registration: This study is approved by the Danish Inspectorate of Animal Experiments (2010/561-1837).

No MeSH data available.


Related in: MedlinePlus

Apoptosis quantification. The mean value of apoptotic cell profiles per square millimeter in the two arbitrary sections from each kidney is used. Black horizontal lines are medians, and whiskers are interquartile range. Within the groups, p values from Wilcoxon signed rank test are shown, and between the groups p values from Mann–Whitney rank sum test are shown. IK, ischemic kidney; CK, control kidney.
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Figure 5: Apoptosis quantification. The mean value of apoptotic cell profiles per square millimeter in the two arbitrary sections from each kidney is used. Black horizontal lines are medians, and whiskers are interquartile range. Within the groups, p values from Wilcoxon signed rank test are shown, and between the groups p values from Mann–Whitney rank sum test are shown. IK, ischemic kidney; CK, control kidney.

Mentions: A representative picture of the caspase-3-positive staining used for apoptotic cell counts is shown in Figure 4. For quantification, a mean of 104 randomly selected non-overlapping fields of view were used per section. The majority of the apoptotic cells were located in the tubules, but the apoptotic cells were also found in the glomeruli and in the lumen of the tubules. Quantification of caspase-3-positive cell profiles is presented in Figure 5. The degree of caspase-3-active cell profiles presented as median (IQR) were significantly higher in the kidneys subjected to ischemia (IK120 0.86 (0.47,1.36); IK240 1.56 (0.83,2.89)) compared to the control kidney (CK120 0.07 (0.04,0.13), CK240 0.06 (0.00,0.11)), indicating a higher degree of ongoing apoptosis in the affected organ. We found a significant higher degree of apoptotic cell profiles in the sections from the post ischemic kidneys in WI240 compared to WI120. All these differences were statistically significant (p < 0.05).


Cell injury after ischemia and reperfusion in the porcine kidney evaluated by radiolabelled microspheres, sestamibi, and lactadherin.

Pedersen SS, Keller AK, Nielsen MK, Jespersen B, Falborg L, Rasmussen JT, Heegaard CW, Rehling M - EJNMMI Res (2013)

Apoptosis quantification. The mean value of apoptotic cell profiles per square millimeter in the two arbitrary sections from each kidney is used. Black horizontal lines are medians, and whiskers are interquartile range. Within the groups, p values from Wilcoxon signed rank test are shown, and between the groups p values from Mann–Whitney rank sum test are shown. IK, ischemic kidney; CK, control kidney.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3750402&req=5

Figure 5: Apoptosis quantification. The mean value of apoptotic cell profiles per square millimeter in the two arbitrary sections from each kidney is used. Black horizontal lines are medians, and whiskers are interquartile range. Within the groups, p values from Wilcoxon signed rank test are shown, and between the groups p values from Mann–Whitney rank sum test are shown. IK, ischemic kidney; CK, control kidney.
Mentions: A representative picture of the caspase-3-positive staining used for apoptotic cell counts is shown in Figure 4. For quantification, a mean of 104 randomly selected non-overlapping fields of view were used per section. The majority of the apoptotic cells were located in the tubules, but the apoptotic cells were also found in the glomeruli and in the lumen of the tubules. Quantification of caspase-3-positive cell profiles is presented in Figure 5. The degree of caspase-3-active cell profiles presented as median (IQR) were significantly higher in the kidneys subjected to ischemia (IK120 0.86 (0.47,1.36); IK240 1.56 (0.83,2.89)) compared to the control kidney (CK120 0.07 (0.04,0.13), CK240 0.06 (0.00,0.11)), indicating a higher degree of ongoing apoptosis in the affected organ. We found a significant higher degree of apoptotic cell profiles in the sections from the post ischemic kidneys in WI240 compared to WI120. All these differences were statistically significant (p < 0.05).

Bottom Line: In WI240, the uptake of microspheres was severely reduced in both groups (17% ± 11% and 27% ± 17%).GFR was severely reduced in the post ischemic kidney in both groups.In both groups, the uptake of lactadherin was reduced (41% ± 8%, 17% ± 13%) but not different from the uptake of (153)Gd microspheres.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Clinical Medicine, Aarhus University, Aarhus, Denmark ; Department of Clinical Physiology and Nuclear Medicine, Aarhus University Hospital, Skejby, Denmark.

ABSTRACT

Background: The purpose of the present study was to quantify renal cell injury after ischemia and reperfusion in a pig model using (99m)Tc-lactadherin as a marker of apoptosis and (99m)Tc-sestamibi as a marker of mitochondrial dysfunction.

Methods: Thirty-four pigs were randomized into unilateral renal warm ischemia of 120 (WI120) or 240 min (WI240). The glomerular filtration rate (GFR) was calculated by renal clearance of (51)Cr-ethylenediaminetetraacetic acid, and apoptosis was quantified by immunohistochemical detection of caspase-3. After 240 min of reperfusion, intravenous (99m)Tc-lactadherin or (99m)Tc-sestamibi was injected simultaneously with (153)Gd microspheres into the aorta. Ex-vivo static planar images of the kidneys were acquired for determination of the differential renal function of tracer distribution using a gamma camera.

Results: In WI120, there was no significant difference in the uptake of microspheres in the ischemic and contralateral normal kidney indicating adequate perfusion (uptake in ischemic kidney relative to the sum of uptake in both kidneys; 46% ± 12% and 51% ± 5%). In WI240, the uptake of microspheres was severely reduced in both groups (17% ± 11% and 27% ± 17%). GFR was severely reduced in the post ischemic kidney in both groups. In both groups, the uptake of lactadherin was reduced (41% ± 8%, 17% ± 13%) but not different from the uptake of (153)Gd microspheres. Caspase-3-positive cell profiles were increased in the post-ischemic kidneys (p < 0.001) and increased as the length of ischemia increased (p = 0.003). In both WI120 and WI240, the amount of (99m)Tc-sestamibi in the ischemic kidney was significantly lower than the amount of (153)Gd microspheres (40 ± 5 versus 51 ± 5 and 20 ± 11 versus 27 ± 17; p < 0.05).

Conclusions: In an established pig model with unilateral renal warm ischemia, we found significantly reduced (99m)Tc-sestamibi uptake relative to perfusion in the kidneys exposed to ischemia indicating a potential ability to detect renal ischemic and reperfusion injuries. However, apoptosis was not detected using (99m)Tc-lactadherin in the post-ischemic kidneys despite increased number of caspase-3-positive cell profiles.

Trial registration: This study is approved by the Danish Inspectorate of Animal Experiments (2010/561-1837).

No MeSH data available.


Related in: MedlinePlus