Figure 4
From: eNAMPT neutralization reduces preclinical ARDS severity via rectified NFkB and Akt/mTORC2 signaling
eNAMPT-neutralizing strategies attenuate porcine lung injury in preclinical “two-hit” septic shock/VILI. (A, B) Increased lung tissue NAMPT expression (as described in Fig. 1) in the “two-hit” preclinical porcine septic shock/VILI pigs compared to controls. *p < 0.001. (C) Western blot analysis confirmed significantly higher NAMPT expression in porcine septic shock/VILI pig lung homogenates (lanes 3–5) compared to controls (first 2 lanes). The eNAMPT-neutralizing mAb reduced NAMPT immunoreactivity in lung homogenates (lanes 6–8) (*p < 0.05 control vs. untreated LPS/VILI; **p < 0.05 mAb LPS/VILI vs. untreated LPS/VILI). (D–G) The eNAMPT mAb dramatically reduced H&E histologic and BAL evidence of severe inflammatory lung injury captured by quantification of injury (Image J Software) (D, E) and by reductions in BAL protein (F), and BAL total PMNs (G) (n = 6 pigs/group) (*p < 0.05 control vs. untreated LPS/VILI; **p < 0.05 mAb LPS/VILI vs. untreated LPS/VILI). (H) Serum lactic acid levels were obtained hourly and were significantly increased beginning 1 h after the onset of sepsis/VILI exposure. Lactate levels remained elevated each hour with a significant increase beginning at 9 h compared to 1–8 h. The humanized eNAMPT mAb significantly attenuated serum lactate increases over the 9–12 h time frame with values similar to control animals. (*p < 0.05 control vs. untreated LPS/VILI; **p < 0.05 untreated LPS/VILI 1–8 h vs. untreated LPS/VILI 9–12 h). ***p < 0.05 mAb LPS/VILI 9–12 h vs. untreated LPS/VILI 9–12 h). (I, J) Significant decreases in static and dynamic compliance, reflections of lung stiffness were observed in the initial 4 h period of exposure to sepsis/VILI, further decreasing and stabilizing through 9–12 h. Pigs receiving the eNAMPT neutralizing mAb at 2 h exhibited static and dynamic compliance values that were significantly and continuously improved over the 5–12 h period (p < 0.05) consistent with improved pulmonary/ventilatory physiologic indices. See Supplemental Table 1 and Supplemental Fig. 1.
