![Figure 2. DNase I treatment significantly reduces NETs in TNF-α–stimulated SCD mice, protects SCD mice from NET-associated hypothermia, and prolongs their survival by reducing acute lung injury. (A) Quantification of NETs in the lungs of TNF-α–stimulated SCD mice pretreated with vehicle (white bar, n = 3) or DNase I (red bar, n = 7). Results presented are the average values of independent experiments with sex- and age-matched mice (mean ± SEM, ***P < .001). (B) Quantification of NET biomarker, plasma DNA, of TNF-α–stimulated SCD mice pretreated with vehicle (white circle, n = 6) or pretreated with DNase I (red circle, n = 10) (Mann-Whitney test, median [IQR]). (C) Reduction in rectal temperature of TNF-α–stimulated SA mice (gray circle, n = 10), PBS-treated SCD mice (green circle, n = 3), and TNF-α–stimulated SCD mice (white circle, n = 10; mean ± SEM, *P < .05, ****P < .0001). (D) Reduction in rectal temperature of TNF-α–stimulated SCD mice, pretreated with vehicle (white circle, n = 6) or DNase I (red circle, n = 11; mean ± SEM, *P < .05). (E) Relationship between decrease in body temperature and number of NETs presented in the lungs of SA and SCD mice (r = 0.79, P < .0001, different color circles represent different treatments as stated in Figure 2C-D). (F) Kaplan-Meier survival curves after TNF-α treatment and surgical trauma in vehicle-treated (black line, n = 5) and DNase I–treated (red line, n = 7) groups (log-rank test, *P < .05). (G) Representative histology of lungs from vehicle-infused (arrows indicate the fibrillar appearance of alveolar walls) and DNase I–infused SCD mice after TNF-α administration and surgical procedure. Scale bar, 10 μm. Images were captured using Zeiss Axio Observer Z1 microscope equipped with Zeiss Axiocam HRc camera (color) and a 40× oil immersion objective. (H) (Top) Total protein (left) and IgM concentration (right) in BAL fluid of vehicle- (white circle, n = 7) and DNase I–treated (red circle, n = 7) SCD mice and vehicle-treated SA mice (gray circle, n = 4) after TNF-α administration and surgical procedure (mean ± SEM, *P < .05, **P < .01). (Bottom) MPO activity per gram of tissue in lung homogenates (left) and in cell-free BAL fluid (right) of vehicle- (white circle, n = 7) and DNase I–treated (red circle, n = 7) SCD mice and vehicle-treated SA mice (gray circle, n = 4) after TNF-α administration and surgical procedure (mean ± SEM, * P < .05, **P < .01).](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/123/24/10.1182_blood-2013-10-529982/4/3818f2.jpeg?Expires=1722768312&Signature=KOzIoI0uY-yuhKmFhrv~wdrzuw8sQRlL~zjFzZqfNN54bMa~bQymICtXSf60u3l1lJNpnT2mAhkVnE9bCnutTb6VaQcXvEyGVrBoSipsTH~c1KH81Ag119EBE5xw2Fm6fyWoV51Zg77BSQMri6bmTc55zl0bpbqijecqE3zZdTpKLjYK5HZ3Ja4VU2NfG0cIpF3DEyc9wgYhkjwhFN16mralMyAxeOOuyD1id~vhPfwpM0SyZG5D1oY9045vv~YvNxJgW6t9fn-9B~XUhxAO0LbEoZ3gJQEBApPnOM0VqZEpVrxDIaursKNerDd7byRfrHz8LxCYNK0VD-bnmk-tdw__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
DNase I treatment significantly reduces NETs in TNF-α–stimulated SCD mice, protects SCD mice from NET-associated hypothermia, and prolongs their survival by reducing acute lung injury. (A) Quantification of NETs in the lungs of TNF-α–stimulated SCD mice pretreated with vehicle (white bar, n = 3) or DNase I (red bar, n = 7). Results presented are the average values of independent experiments with sex- and age-matched mice (mean ± SEM, ***P < .001). (B) Quantification of NET biomarker, plasma DNA, of TNF-α–stimulated SCD mice pretreated with vehicle (white circle, n = 6) or pretreated with DNase I (red circle, n = 10) (Mann-Whitney test, median [IQR]). (C) Reduction in rectal temperature of TNF-α–stimulated SA mice (gray circle, n = 10), PBS-treated SCD mice (green circle, n = 3), and TNF-α–stimulated SCD mice (white circle, n = 10; mean ± SEM, *P < .05, ****P < .0001). (D) Reduction in rectal temperature of TNF-α–stimulated SCD mice, pretreated with vehicle (white circle, n = 6) or DNase I (red circle, n = 11; mean ± SEM, *P < .05). (E) Relationship between decrease in body temperature and number of NETs presented in the lungs of SA and SCD mice (r = 0.79, P < .0001, different color circles represent different treatments as stated in Figure 2C-D). (F) Kaplan-Meier survival curves after TNF-α treatment and surgical trauma in vehicle-treated (black line, n = 5) and DNase I–treated (red line, n = 7) groups (log-rank test, *P < .05). (G) Representative histology of lungs from vehicle-infused (arrows indicate the fibrillar appearance of alveolar walls) and DNase I–infused SCD mice after TNF-α administration and surgical procedure. Scale bar, 10 μm. Images were captured using Zeiss Axio Observer Z1 microscope equipped with Zeiss Axiocam HRc camera (color) and a 40× oil immersion objective. (H) (Top) Total protein (left) and IgM concentration (right) in BAL fluid of vehicle- (white circle, n = 7) and DNase I–treated (red circle, n = 7) SCD mice and vehicle-treated SA mice (gray circle, n = 4) after TNF-α administration and surgical procedure (mean ± SEM, *P < .05, **P < .01). (Bottom) MPO activity per gram of tissue in lung homogenates (left) and in cell-free BAL fluid (right) of vehicle- (white circle, n = 7) and DNase I–treated (red circle, n = 7) SCD mice and vehicle-treated SA mice (gray circle, n = 4) after TNF-α administration and surgical procedure (mean ± SEM, * P < .05, **P < .01).
