Figure 5
From: Acid Sphingomyelinase Promotes Endothelial Stress Response in Systemic Inflammation and Sepsis
Detrimental role of SMPD1 in sepsis-induced endothelial stress response Semi-confluent HMECs (1 × 104) were incubated for 24 h with serum (5%) obtained from healthy individuals, from patients with sepsis (n = 6) or proinflammatory cytokines. Co-incubation with (A) SMPD1 specific siRNA (5 nmol/L, preincubation for 48 h) or (B) SMPD1 direct inhibitor amino bisphosphonate (0.1 µmol/L) prevented ADAMTS13 down-regulation as normalized to reference transcript GAPDH. Stimulation of HMECs with (C) isolated human acid sphingomyelinase resulted in significant LDH release into the supernatant, whereas (D) amino bisphophonates abrogated LDH release following cytokine stimulation. The boxes indicate the interquartile range, divided by the median (central horizontal line) as analyzed in at least four independent experiments.