Levosimendan attenuates multiple organ injury and improves survival in peritonitis-induced septic shock: studies in a rat model

Abstract

Introduction: The aim of this study was to investigate the effects of levosimendan on rodent septic shock induced by cecal ligation and puncture (CLP).

Methods: Three hours after peritonitis-induced sepsis, male Wistar rats were randomly assigned to receive an intravenous infusion of levosimendan (1.2 μg/kg/min for 10 min and then 0.3 μg/kg/min for 6 h) or an equivalent volume of saline and vehicle (5% dextrose) solution.

Results: The levosimendan-treated CLP animals had significantly higher arterial pressure and lower biochemical indices of liver and kidney dysfunction compared to the CLP animals (P < 0.05). Plasma interleukin-1β, nitric oxide and organ superoxide levels in the levosimendan-treated CLP group were less than those in CLP rats treated with vehicle (P < 0.05). In addition, the inducible nitric oxide synthase (iNOS) in lung and caspase-3 expressions in spleen were significantly lower in the levosimendan-treated CLP group (P < 0.05). The administration of CLP rats with levosimendan was associated with significantly higher survival (61.9% vs. 40% at 18 h after CLP, P < 0.05). At postmortem examination, the histological changes and neutrophil filtration index in liver and lung were significantly attenuated in the levosimendan-treated CLP group (vs. CLP group, P < 0.05).

Conclusions: In this clinically relevant model of septic shock induced by fecal peritonitis, the administration of levosimendan had beneficial effects on haemodynamic variables, liver and kidney dysfunction, and metabolic acidosis. (1) Lower levels of interleukin-1β, nitric oxide and superoxide, (2) attenuation of iNOS and caspase-3 expressions, and (3) decreases of neutrophil infiltration by levosimendan in peritonitis-induced sepsis animals suggest that anti-inflammation and anti-apoptosis effects of levosimendan contribute to prolonged survival.

Figure 1

Progress of plasma (A) alanine aminotransferase (ALT), (B) aspartate aminotransferase (AST), (C) lactate dehydrogenase (LDH), (D) blood urea nitrogen (BUN), (E) creatinine, and (F) blood glucose during the experimental period. Depicted are the changes in plasma biochemical parameters in different groups of sham operation (SOP, n = 10), SOP plus levosimendan administration (SOP + LS, n = 6), cecal ligation and puncture (CLP, n = 10), CLP plus vehicle treatment (CLP + Veh, n = 6) and CLP plus levosimendan administration (CLP + LS, n = 12). Data are expressed as mean ± standard error of the mean. *P <0.05, all groups versus SOP. ^† P <0.05, CLP + LS versus CLP + Veh.

Figure 2

Progress of plasma (A) nitric oxide (NO) and (B) interleukin-1β (IL-1β) levels during the experimental period. Depicted are the changes of plasma NO and IL-1β levels in different groups of sham operation (SOP, n = 10), SOP plus Levosimendan administration (SOP + LS, n = 6), cecal ligation and puncture (CLP, n = 10), CLP plus vehicle treatment (CLP + Veh, n = 6) and CLP plus Levosimendan administration (CLP + LS, n = 12). Data are expressed as mean ± standard error of the mean. *P <0.05, all groups versus SOP; ^† P <0.05, CLP + LS versus CLP + Veh.

Figure 3

Superoxide levels of aorta, liver, spleen and pancreas at 18 h after surgery. Depicted are the changes in different groups of sham operation (SOP, n = 10), SOP plus Levosimendan administration (SOP + LS, n = 6), cecal ligation and puncture (CLP, n = 10), CLP plus vehicle treatment (CLP + Veh, n = 6) and CLP plus Levosimendan administration (CLP + LS, n = 12). Data are expressed as mean ± standard error of the mean. *P <0.05, all groups versus SOP, ^† P <0.05, CLP + LS versus CLP + Veh.

Figure 4

Expressions of (A) lung inducible nitric oxide (NO) synthase (iNOS) and (B) spleen-cleaved caspase 3. Depicted are the expressions of lung iNOS and spleen-cleaved caspase 3 at the end of the study in different groups of sham operation (SOP, n = 3), SOP plus Levosimendan administration (SOP + LS, n = 3), cecal ligation and puncture (CLP, n = 5), CLP plus vehicle treatment (CLP + Veh, n = 5) and CLP plus Levosimendan administration (CLP + LS, n = 5). Data are expressed as mean ± standard error of the mean. *P <0.05, all groups versus SOP, ^† P <0.05, CLP + LS versus CLP + Veh.

Figure 5

Histopathological studies of lung and liver. Light microscopy showed lung and liver sections of rats in groups of (A) sham operation, (B) cecal ligation and puncture (CLP), and (C) CLP plus Levosimendan administration. Sections were stained with haematoxylin and eosin. Arrows represent polymorphonuclear neutrophil infiltration in lung and liver. Each is shown at 400 × (original magnification).

Figure 6

Histopathological studies of spleen. Light microscopy showed spleen sections of rats in groups of (A) sham operation, (B) cecal ligation and puncture, and (C) CLP plus Levosimendan administration. Sections were stained with haematoxylin and eosin. Arrows represent apoptosis in the spleen. Each is shown at 400 × (original magnification).