Cisatracurium attenuates LPS-induced modulation of MMP3 and junctional protein expression in human microvascular endothelial cells

Abstract

Acute respiratory distress syndrome (ARDS) is a life-threatening form of acute lung injury (ALI) associated with hypoxemic lung damage and inflammation. Matrix metalloproteinase protein-3 (MMP3 or Stromelysin-1) is known to promote vascular injury in ALI/ARDS. Cisatracurium, a nicotinic neuromuscular blocker, is used in ARDS patients to decrease mechanical ventilator dyssynchrony, increase oxygenation, and improve mortality. However, the magnitude and the underlying mechanisms of these potential benefits of cisatracurium remains unclear. We investigated the effect of cisatracurium on lipopolysaccharide-induced MMP3 expression in human microvascular endothelial cells. In our results, cisatracurium treatment significantly decreased LPS-induced MMP3 expression and increased expression of cell junction proteins such as vascular endothelial cadherin (VE-cadherin) and claudin-5.

Keywords: Cisatracurium; MMP3; cell junction; lipopolysaccharide; stromelysin-1.

Figure 1.. Cisatracurium inhibited LPS-induced increase in MMP3 expression and Src phosphorylation.

(A-B) Representative Western blot images and a bar graph with band densitometry analysis indicating increased MMP3 expression in HMECs with LPS treatment and its reversal by co-treatment with cisatracurium after 24 hours of incubation. (C) Bar graph with band densitometry analysis indicating increased pY416Src expression in HMECs with LPS treatment and its reversal by co-treatment with cisatracurium after 24 hours of incubation. (D) Bar graph with band densitometry analysis indicating a modest increase in pP38MAPK expression in HMECs with LPS treatment and its partial inhibition by co-treatment with a very low dose of cisatracurium after 24 hours of incubation. Data are shown as Mean + SD.

Figure 2.. Cisatracurium prevented LPS-induced loss of VE-cadherin and claudin-5 in HMECs.

(A) Representative Western blot images showing reduced VE-cadherin and claudin-5 expressions in HMECs with LPS treatment and its reversal by co-treatment with cisatracurium after 24 hours of incubation. (B) Bar graph with band densitometry analysis indicating reduced VE-cadherin expression in HMECs with LPS treatment and its reversal by co-treatment with cisatracurium after 24 hours of incubation. (C) Bar graph with band densitometry analysis indicating reduced claudin-5 expression in HMECs with LPS treatment and reversal by co-treatment with cisatracurium after 24 hours of incubation. Data are shown as Mean + SD.

Figure 3.

Representative images of VE-cadherin staining in human microvascular endothelial cells post LPS treatment alone, or in combination with cisatracurium at 24 hours.