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A cardiopulmonary bypass with deep hypothermic circulatory arrest rat model for the investigation of the systemic inflammation response and induced organ damage.

Engels M, Bilgic E, Pinto A, Vasquez E, Wollschläger L, Steinbrenner H, Kellermann K, Akhyari P, Lichtenberg A, Boeken U - J Inflamm (Lond) (2014)

Bottom Line: Results were compared to animals (n = 6) which did not undergo CPB.Activation of MAPK and induction of heat shock proteins occurred in an organ-specific manner with most pronounced effects in heart, lungs and kidneys.This model might be suitable to test the efficacy of therapeutics applied in major heart surgery with and without DHCA.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Cardiovascular Surgery, Faculty of Medicine, Heinrich Heine University, Moorenstr. 5, Duesseldorf, D - 40225, Germany ; Institute for Biochemistry and Molecular Biology I, Heinrich Heine University, Duesseldorf, Germany.

ABSTRACT

Background: Cardiopulmonary bypass (CPB) is a commonly used technique in cardiac surgery. CPB is however associated with a strong induction of systemic inflammatory response syndrome (SIRS) which in conjunction with ischemia and reperfusion may lead to multiple organ failure. The aim of the study was to establish and characterize a CPB rat model incorporating deep hypothermic circulatory arrest with a specific focus on the extent of the inflammatory reactions and organ damage as a groundwork for novel therapeutics against SIRS and I/R induced organ injury.

Materials and methods: Male Wistar rats (n = 6) were cannulated for CPB, connected to a heart-lung-machine (HLM) and cooled to a temperature of 16°C before they underwent 45 minutes of deep hypothermic circulatory arrest with global ischaemia. Arrest was followed by rewarming and 60 minutes of reperfusion. Haemodynamic and vital parameters were recorded throughout the CPB procedure. Only animals displaying sinus rhythm throughout reperfusion were utilized for analysis. Rats were euthanized and tissue samples were harvested. Blood gas analysis was performed and blood samples were taken. Induction of organ damage was examined by analysis of protein levels and phosphorylation status of kinases and stress proteins. Results were compared to animals (n = 6) which did not undergo CPB.

Results: CPB induced leucocytosis and an increase of interleukin-6 and TNF-α plasma values indicating an inflammatory response. Markers of tissue damage and dysfunction, such as troponin T, creatinine and AST were elevated. Phosphorylation of STAT3 was induced in all examined organs. Activation of MAPK and induction of heat shock proteins occurred in an organ-specific manner with most pronounced effects in heart, lungs and kidneys.

Conclusions: The presented CPB rat model shows the induction of SIRS and activation of specific signalling cascades. SIRS seems not to be provoked during DHCA and is elicited mainly during reperfusion. This model might be suitable to test the efficacy of therapeutics applied in major heart surgery with and without DHCA.

No MeSH data available.


Related in: MedlinePlus

Plasma IL-6 and TNF-α Levels. IL-6 (A) and TNF-α (B) were measured in healthy animals and in I/R animals before CPB (T1), after 35 minutes of cooling (T2) and after 60 minutes of reperfusion (T3). Values are given as pg/ml. Data are presented as mean ± SEM of six animals. Significance was determined by one way ANOVA with Tukey-test post analysis. A: T5-values were significantly higher than T2-, T1- and basal animal-values; B: T5-values were significantly higher than T1- and basal animal-values.
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Figure 3: Plasma IL-6 and TNF-α Levels. IL-6 (A) and TNF-α (B) were measured in healthy animals and in I/R animals before CPB (T1), after 35 minutes of cooling (T2) and after 60 minutes of reperfusion (T3). Values are given as pg/ml. Data are presented as mean ± SEM of six animals. Significance was determined by one way ANOVA with Tukey-test post analysis. A: T5-values were significantly higher than T2-, T1- and basal animal-values; B: T5-values were significantly higher than T1- and basal animal-values.

Mentions: IL-6 increase is associated with reperfusion and induces a variety of downstream events, e.g. cardioprotection by JAK/STAT signalling during CPB. We therefore determined the plasma IL-6 and TNF-α levels at T1, T2 and T5. Rewarming and reperfusion (T5) following DHCA led to a dramatic increase of IL-6 in all animals, causing significantly elevated values as compared to time points prior to DHCA or as compared to values observed in healthy animals (Figure 3A). Noteworthy, IL-6-levels of the T1 and T2 samples all lay under the detection level (according to the manufacturer at 19 pg/ml). TNF-α-levels were also significantly elevated after reperfusion as compared to prior time points and to healthy animals. In contrast to the IL-6-levels, TNF-α-levels were already elevated after 25 minutes of cooling (Figure 3B). Therewith the present study could demonstrate that I/R injury as applied in the presented model leads to an increase of the pro-inflammatory cytokines IL-6 and TNF-α.


A cardiopulmonary bypass with deep hypothermic circulatory arrest rat model for the investigation of the systemic inflammation response and induced organ damage.

Engels M, Bilgic E, Pinto A, Vasquez E, Wollschläger L, Steinbrenner H, Kellermann K, Akhyari P, Lichtenberg A, Boeken U - J Inflamm (Lond) (2014)

Plasma IL-6 and TNF-α Levels. IL-6 (A) and TNF-α (B) were measured in healthy animals and in I/R animals before CPB (T1), after 35 minutes of cooling (T2) and after 60 minutes of reperfusion (T3). Values are given as pg/ml. Data are presented as mean ± SEM of six animals. Significance was determined by one way ANOVA with Tukey-test post analysis. A: T5-values were significantly higher than T2-, T1- and basal animal-values; B: T5-values were significantly higher than T1- and basal animal-values.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4231204&req=5

Figure 3: Plasma IL-6 and TNF-α Levels. IL-6 (A) and TNF-α (B) were measured in healthy animals and in I/R animals before CPB (T1), after 35 minutes of cooling (T2) and after 60 minutes of reperfusion (T3). Values are given as pg/ml. Data are presented as mean ± SEM of six animals. Significance was determined by one way ANOVA with Tukey-test post analysis. A: T5-values were significantly higher than T2-, T1- and basal animal-values; B: T5-values were significantly higher than T1- and basal animal-values.
Mentions: IL-6 increase is associated with reperfusion and induces a variety of downstream events, e.g. cardioprotection by JAK/STAT signalling during CPB. We therefore determined the plasma IL-6 and TNF-α levels at T1, T2 and T5. Rewarming and reperfusion (T5) following DHCA led to a dramatic increase of IL-6 in all animals, causing significantly elevated values as compared to time points prior to DHCA or as compared to values observed in healthy animals (Figure 3A). Noteworthy, IL-6-levels of the T1 and T2 samples all lay under the detection level (according to the manufacturer at 19 pg/ml). TNF-α-levels were also significantly elevated after reperfusion as compared to prior time points and to healthy animals. In contrast to the IL-6-levels, TNF-α-levels were already elevated after 25 minutes of cooling (Figure 3B). Therewith the present study could demonstrate that I/R injury as applied in the presented model leads to an increase of the pro-inflammatory cytokines IL-6 and TNF-α.

Bottom Line: Results were compared to animals (n = 6) which did not undergo CPB.Activation of MAPK and induction of heat shock proteins occurred in an organ-specific manner with most pronounced effects in heart, lungs and kidneys.This model might be suitable to test the efficacy of therapeutics applied in major heart surgery with and without DHCA.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Cardiovascular Surgery, Faculty of Medicine, Heinrich Heine University, Moorenstr. 5, Duesseldorf, D - 40225, Germany ; Institute for Biochemistry and Molecular Biology I, Heinrich Heine University, Duesseldorf, Germany.

ABSTRACT

Background: Cardiopulmonary bypass (CPB) is a commonly used technique in cardiac surgery. CPB is however associated with a strong induction of systemic inflammatory response syndrome (SIRS) which in conjunction with ischemia and reperfusion may lead to multiple organ failure. The aim of the study was to establish and characterize a CPB rat model incorporating deep hypothermic circulatory arrest with a specific focus on the extent of the inflammatory reactions and organ damage as a groundwork for novel therapeutics against SIRS and I/R induced organ injury.

Materials and methods: Male Wistar rats (n = 6) were cannulated for CPB, connected to a heart-lung-machine (HLM) and cooled to a temperature of 16°C before they underwent 45 minutes of deep hypothermic circulatory arrest with global ischaemia. Arrest was followed by rewarming and 60 minutes of reperfusion. Haemodynamic and vital parameters were recorded throughout the CPB procedure. Only animals displaying sinus rhythm throughout reperfusion were utilized for analysis. Rats were euthanized and tissue samples were harvested. Blood gas analysis was performed and blood samples were taken. Induction of organ damage was examined by analysis of protein levels and phosphorylation status of kinases and stress proteins. Results were compared to animals (n = 6) which did not undergo CPB.

Results: CPB induced leucocytosis and an increase of interleukin-6 and TNF-α plasma values indicating an inflammatory response. Markers of tissue damage and dysfunction, such as troponin T, creatinine and AST were elevated. Phosphorylation of STAT3 was induced in all examined organs. Activation of MAPK and induction of heat shock proteins occurred in an organ-specific manner with most pronounced effects in heart, lungs and kidneys.

Conclusions: The presented CPB rat model shows the induction of SIRS and activation of specific signalling cascades. SIRS seems not to be provoked during DHCA and is elicited mainly during reperfusion. This model might be suitable to test the efficacy of therapeutics applied in major heart surgery with and without DHCA.

No MeSH data available.


Related in: MedlinePlus