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Apoptotic cell administration is detrimental in murine renal ischaemia reperfusion injury.

Hesketh EE, Kluth DC, Hughes J - J Inflamm (Lond) (2014)

Bottom Line: The administration of apoptotic cells (ACs) has been shown to reduce inflammation in various organs including the liver and kidney.Early AC administration prior to severe IRI had no influence on plasma creatinine or ATN severity.These data suggest that AC-derived protection is not translationally relevant for patients with acute kidney injury induced by ischaemic injury.

View Article: PubMed Central - PubMed

Affiliation: MRC Centre for Inflammation Research, Queen's Medical Research Institute, University of Edinburgh, 47 Little France Crescent, Edinburgh, EH16 4TJ UK.

ABSTRACT

Background: Acute kidney injury induced by renal ischaemia reperfusion injury (IRI) is characterised by renal failure, acute tubular necrosis (ATN), inflammation and microvascular congestion. The administration of apoptotic cells (ACs) has been shown to reduce inflammation in various organs including the liver and kidney. This study explored whether AC administration prior to the induction of renal IRI was protective.

Findings: Renal IRI was induced in Balb/c mice by clamping the renal blood vessels for either 20, 24 or 25 minutes to induce mild, moderate or severe kidney dysfunction respectively. Renal function and injury was determined 24 hours following IRI by measurement of plasma creatinine and ATN scoring respectively. ACs were generated from Balb/c thymocytes and classified as either predominantly early or late apoptotic by Annexin-V and propidium iodide staining. Early AC administration prior to severe IRI had no influence on plasma creatinine or ATN severity. In contrast, administration of early or late ACs significantly worsened renal function in mice with mild or moderate renal IRI, respectively, compared to PBS treated controls, though ATN scores were comparable. Despite ACs exerting pro-coagulant effects, the worsening of renal function was not secondary to increased microvascular congestion, inferred by fibrin and platelet (CD41) deposition, or inflammation, assessed by neutrophil infiltration.

Conclusions: Despite the AC-derived protection demonstrated in other organs, ACs do not protect mice from renal IRI. ACs may in fact further impair renal function depending on injury severity. These data suggest that AC-derived protection is not translationally relevant for patients with acute kidney injury induced by ischaemic injury.

No MeSH data available.


Related in: MedlinePlus

Neutrophil infiltration occurred following ischaemia but was unaffected by AC administration. Renal IRI was induced in 8-week old male Balb/c mice by a right nephrectomy and ischaemia induced by occluding the left renal pedicle for 20 (mild ischaemia), 24 (moderate) or 25 mins (severe). Sham mice underwent a laparotomy only. Either PBS or 20×106 early or late ACs were administered 24 hr prior to renal IRI. Mice were sacrificed 24 hr following IRI. (A) Representative images of the outer stripe of the outer medulla in non-injured and ischaemic kidney sections following Gr1 staining. In ischaemic kidneys neutrophil infiltration was observed in areas of necrosis. (Magnification: ×200; Scale Bar: 50 μM) (B) Scoring of Gr1 staining (expressed as Gr1+ area) demonstrates that neutrophil infiltration remained similar between PBS and AC treated mice with severe, moderate and mild ischaemic injury. Grey circle symbol = Non-injured kidney Grey square symbol = Ischaemic kidney. Data expressed as mean ± SEM and analysed by two-way ANOVA. ns = non-significant. Sham (n = 4), PBS (n = 7–8), AC (n = 8).
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Fig5: Neutrophil infiltration occurred following ischaemia but was unaffected by AC administration. Renal IRI was induced in 8-week old male Balb/c mice by a right nephrectomy and ischaemia induced by occluding the left renal pedicle for 20 (mild ischaemia), 24 (moderate) or 25 mins (severe). Sham mice underwent a laparotomy only. Either PBS or 20×106 early or late ACs were administered 24 hr prior to renal IRI. Mice were sacrificed 24 hr following IRI. (A) Representative images of the outer stripe of the outer medulla in non-injured and ischaemic kidney sections following Gr1 staining. In ischaemic kidneys neutrophil infiltration was observed in areas of necrosis. (Magnification: ×200; Scale Bar: 50 μM) (B) Scoring of Gr1 staining (expressed as Gr1+ area) demonstrates that neutrophil infiltration remained similar between PBS and AC treated mice with severe, moderate and mild ischaemic injury. Grey circle symbol = Non-injured kidney Grey square symbol = Ischaemic kidney. Data expressed as mean ± SEM and analysed by two-way ANOVA. ns = non-significant. Sham (n = 4), PBS (n = 7–8), AC (n = 8).

Mentions: With increased microvascular congestion not explaining this surprising result it was speculated that AC administration might have increased immune cell infiltration thereby elevating pro-inflammatory responses post renal IRI. Neutrophils rapidly accumulate after renal ischaemic injury [3]. They adhere to the microvascular endothelium where they may obstruct the renal microvasculature and potentially damage tubular cells by releasing reactive oxygen species [20]. To gain an insight into the inflammatory status, neutrophil infiltration was determined following IHC staining for Gr1. Neutrophil infiltration was observed in the OSOM of all ischaemic kidneys of PBS and AC treated mice (Figure 5A), however this remained equivalent in all AC treated mice and their corresponding PBS controls (Figure 5B).Figure 5


Apoptotic cell administration is detrimental in murine renal ischaemia reperfusion injury.

Hesketh EE, Kluth DC, Hughes J - J Inflamm (Lond) (2014)

Neutrophil infiltration occurred following ischaemia but was unaffected by AC administration. Renal IRI was induced in 8-week old male Balb/c mice by a right nephrectomy and ischaemia induced by occluding the left renal pedicle for 20 (mild ischaemia), 24 (moderate) or 25 mins (severe). Sham mice underwent a laparotomy only. Either PBS or 20×106 early or late ACs were administered 24 hr prior to renal IRI. Mice were sacrificed 24 hr following IRI. (A) Representative images of the outer stripe of the outer medulla in non-injured and ischaemic kidney sections following Gr1 staining. In ischaemic kidneys neutrophil infiltration was observed in areas of necrosis. (Magnification: ×200; Scale Bar: 50 μM) (B) Scoring of Gr1 staining (expressed as Gr1+ area) demonstrates that neutrophil infiltration remained similar between PBS and AC treated mice with severe, moderate and mild ischaemic injury. Grey circle symbol = Non-injured kidney Grey square symbol = Ischaemic kidney. Data expressed as mean ± SEM and analysed by two-way ANOVA. ns = non-significant. Sham (n = 4), PBS (n = 7–8), AC (n = 8).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Fig5: Neutrophil infiltration occurred following ischaemia but was unaffected by AC administration. Renal IRI was induced in 8-week old male Balb/c mice by a right nephrectomy and ischaemia induced by occluding the left renal pedicle for 20 (mild ischaemia), 24 (moderate) or 25 mins (severe). Sham mice underwent a laparotomy only. Either PBS or 20×106 early or late ACs were administered 24 hr prior to renal IRI. Mice were sacrificed 24 hr following IRI. (A) Representative images of the outer stripe of the outer medulla in non-injured and ischaemic kidney sections following Gr1 staining. In ischaemic kidneys neutrophil infiltration was observed in areas of necrosis. (Magnification: ×200; Scale Bar: 50 μM) (B) Scoring of Gr1 staining (expressed as Gr1+ area) demonstrates that neutrophil infiltration remained similar between PBS and AC treated mice with severe, moderate and mild ischaemic injury. Grey circle symbol = Non-injured kidney Grey square symbol = Ischaemic kidney. Data expressed as mean ± SEM and analysed by two-way ANOVA. ns = non-significant. Sham (n = 4), PBS (n = 7–8), AC (n = 8).
Mentions: With increased microvascular congestion not explaining this surprising result it was speculated that AC administration might have increased immune cell infiltration thereby elevating pro-inflammatory responses post renal IRI. Neutrophils rapidly accumulate after renal ischaemic injury [3]. They adhere to the microvascular endothelium where they may obstruct the renal microvasculature and potentially damage tubular cells by releasing reactive oxygen species [20]. To gain an insight into the inflammatory status, neutrophil infiltration was determined following IHC staining for Gr1. Neutrophil infiltration was observed in the OSOM of all ischaemic kidneys of PBS and AC treated mice (Figure 5A), however this remained equivalent in all AC treated mice and their corresponding PBS controls (Figure 5B).Figure 5

Bottom Line: The administration of apoptotic cells (ACs) has been shown to reduce inflammation in various organs including the liver and kidney.Early AC administration prior to severe IRI had no influence on plasma creatinine or ATN severity.These data suggest that AC-derived protection is not translationally relevant for patients with acute kidney injury induced by ischaemic injury.

View Article: PubMed Central - PubMed

Affiliation: MRC Centre for Inflammation Research, Queen's Medical Research Institute, University of Edinburgh, 47 Little France Crescent, Edinburgh, EH16 4TJ UK.

ABSTRACT

Background: Acute kidney injury induced by renal ischaemia reperfusion injury (IRI) is characterised by renal failure, acute tubular necrosis (ATN), inflammation and microvascular congestion. The administration of apoptotic cells (ACs) has been shown to reduce inflammation in various organs including the liver and kidney. This study explored whether AC administration prior to the induction of renal IRI was protective.

Findings: Renal IRI was induced in Balb/c mice by clamping the renal blood vessels for either 20, 24 or 25 minutes to induce mild, moderate or severe kidney dysfunction respectively. Renal function and injury was determined 24 hours following IRI by measurement of plasma creatinine and ATN scoring respectively. ACs were generated from Balb/c thymocytes and classified as either predominantly early or late apoptotic by Annexin-V and propidium iodide staining. Early AC administration prior to severe IRI had no influence on plasma creatinine or ATN severity. In contrast, administration of early or late ACs significantly worsened renal function in mice with mild or moderate renal IRI, respectively, compared to PBS treated controls, though ATN scores were comparable. Despite ACs exerting pro-coagulant effects, the worsening of renal function was not secondary to increased microvascular congestion, inferred by fibrin and platelet (CD41) deposition, or inflammation, assessed by neutrophil infiltration.

Conclusions: Despite the AC-derived protection demonstrated in other organs, ACs do not protect mice from renal IRI. ACs may in fact further impair renal function depending on injury severity. These data suggest that AC-derived protection is not translationally relevant for patients with acute kidney injury induced by ischaemic injury.

No MeSH data available.


Related in: MedlinePlus