An in�vitro endothelial cell protective effect of secretory leukocyte protease inhibitor against simulated ischaemia/reperfusion injury

Abstract

Endothelial dysfunction is an essential deleterious modulator of ischaemia/reperfusion (I/R) injury. Secretory leukocyte protease inhibitor (SLPI) has demonstrated myocardial protection in cardiac transplantation; however, the effect of SLPI in endothelial I/R injury remains unexplored. In the present study, the effect of recombinant human SLPI (rhSLPI) treatment against endothelial cells (ECs) subjected to simulated I/R injury and the effect of treatment at different time points were determined. Human umbilical vein ECs (HUVECs) were subjected to normoxic or simulated I/R (sI/R) conditions, and rhSLPI at concentrations of 1, 10, 100 and 1,000 ng/ml was added to the cells prior to ischaemia, during ischaemia or at the onset of reperfusion. Endothelial injury and cytoskeleton disruption were assessed, and western blot analysis was conducted. The results revealed that rhSLPI treatment at 1,000 ng/ml significantly increased the HUVEC viability under sI/R injury (P<0.05). In addition, treatment with rhSLPI prior to or during ischaemia markedly attenuated the activity of lactase dehydrogenase compared with that in the sI/R group. In addition, the H2O2-induced reactive oxygen species production was reduced by ~17% upon rhSLPI pretreatment. Endothelial cytoskeleton disruption was also preserved by rhSLPI added prior to the reperfusion period. Furthermore, pretreatment with rhSLPI promoted protein kinase B activation, as well as reduced p38 mitogen-activated protein kinase phosphorylation and B-cell lymphoma 2-associated X protein expression in response to I/R injury. These findings indicated that rhSLPI possesses antioxidant and antiapoptotic properties against endothelial responses to I/R injury. Therefore, the cytoprotective effect of rhSLPI may provide a potential pharmaceutical target to limit endothelial-mediated I/R injury.

Endothelial-Cell-Derived Human Secretory Leukocyte Protease Inhibitor (SLPI) Protects Cardiomyocytes against Ischemia/Reperfusion Injury

“…The current study hypothesized that endothelial-derived rhSLPI could prime the cardiomyocytes and enhance the cardioprotective effect on cardiac cells subjected to in vitro I/R injury. The findings from the current study showed that endothelial-derived rhSLPI showed a greater protective effect than that found using recombinant human SLPI in our previous studies on cardiac resident cells [11,12,13,14].…”

“…Previous studies reported that SLPI acts as an ROS scavenger by reducing intracellular ROS production in many cells during sI/R [11,12,13,14]. To prove that SLPI secreted from EA-SLPI cells reduced intracellular ROS production, in vitro intracellular ROS production was determined under simulated ischemia (sI), hypoxia, and H 2 O 2 .…”

“…The cytoprotective effect of secretory leukocyte protease inhibitor (SLPI) has been reported in different cardiac resident cells, including cardiomyocytes, cardiac fibroblasts, and vascular endothelial cells [11,13,14]. Moreover, SLPI has also been shown to provide cardioprotection against I/R injury [10,12].…”

“…Overexpression of human SLPI in rat cardiac myoblasts (H9c2) or treatment with rhSLPI in both H9c2 and adult rat ventricular myocytes (ARVMs) reduced cell death and injury from an in vitro I/R injury [11,12], as well as decreasing the infarct size [12]. Furthermore, SLPI also protected non-cardiomyocytes such as adult rat cardiac fibroblast cells (ARCFs) and human umbilical vein endothelial cells (HUVECs) from an in vitro simulated I/R (sI/R) injury [13,14]. These reports focused on the direct effect of recombinant human SLPI treatment on individual cardiac resident cells (CM, EC, CFs).…”

SLPI - a Biomarker of Acute Kidney Injury after Open and Endovascular Thoracoabdominal Aortic Aneurysm (TAAA) Repair

“…such as acute lung injury 9,10 . In the setting of oxidative stress, SLPI seems to have antioxidant and cytoprotective properties 11,12 . In a murine model of experimental ischemic AKI, Macrophage Migration Inhibitory Factor-2 (MIF-2) was suggested to exert kidney protection by upregulation of SLPI expression 13 .…”