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Impact of ACE2 Knockout on Elastance and Hemodynamics in Inflammatory Conditions

This study investigates the effects of ACE2 knockout (KO) on elastance and hemodynamic parameters in the presence of inflammatory stimuli such as LPS and Zymosan. Using a series of experiments, we analyze the wet/dry ratio, elastance changes over time, and various angiotensin II responses. Supplementary figures highlight significant differences between wild-type (WT) and ACE2 KO groups in terms of response to treatments and baseline measurements. The findings provide insights into the role of ACE2 in modulating pulmonary and vascular responses during inflammation.

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Impact of ACE2 Knockout on Elastance and Hemodynamics in Inflammatory Conditions

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  1. anti-sense sense a b c 1.0 ACE O kg/ml) 0.8 0.6 ACE2 0.4 Elastance (cmH2 0.2 WT ace2 KO 0.0 WT ace2 KO Supplementary Figure 1

  2. c a * Wet / dry ratio WT, LPS WT, saline ace2 KO, saline ace2 KO, LPS 8 7 6 5 4 b ace2 KO, LPS+Zymosan WT, LPS+Zymosan 150 WT ace2 KO, saline WT, saline 100 % Changes in elastance 50 ace2 KO 0 0 1 2 3 hrs LPS Zymosan Supplementary Figure 2

  3. rhuACE2 2.5 Mut-rhuACE2 Mut-rhuACE2 rhuACE2 2.0 → 175KD 1.5 rhuACE2s Arbitary unit/hr → 83KD → 1.0 62KD Bovine IgG → 48KD 0.5 → 33KD 0 → 1 10 100 (ng/ml) 25KD 9 8 * 7 Wet / dry ratio 6 5 4 rhuACE2 rhuACE2 Mut-rhuACE2 Mut-rhuACE2 acid saline Wild type a b c Supplementary Figure 3

  4. * 4 600 6 600 3 400 AngII (fmol/ml) 4 Ang II (fmol/mg protein) 2 400 Ang II (fmol/mg protein) AngII (fmol/ml) * 200 1 2 200 0 ** ** ** ** 0 0 0 WT, acid ace2 KO, acid WT, control ace2 KO, control WT, acid WT, acid WT, acid ace-/-, acid ace2 KO, acid ace-/-, acid ace2 KO, acid WT, acid, rhuACE2 ace-/-ace2 KO, acid ace-/- ace2 KO, acid a b c d Supplementary Figure 4

  5. 7 * * 6 Wet /dry ratio 5 4 WT, acid WT, saline ace+/-, acid ace-/-, acid ace-/-, saline Supplementary Figure 5

  6. WT, LPS+Zymosan ace+/+ ace2 KO , LPS+Zymosan 150 ace-/- ace2 KO , LPS+Zymosan WT, saline ace-/- ace2 KO, saline 100 % Changes in elastance 50 0 0 1 2 3 hrs LPS Zymosan Supplementary Figure 6

  7. a b * 8 10 7 * Wet / dry ratio 8 Wet / dry ratio 6 * 6 5 4 4 WT, acid WT, saline agtr2-/y, acid agtr1a-/-, acid ace2 KO, saline, vehicle ace2 KO, acid, vehicle ace2 KO, saline, AT1R inhibitor ace2 KO, acid, AT1R inhibitor Supplementary Figure 7

  8. a b WT 12 WT 10 ace2 KO ace2 KO 10 8 8 Pperf (cmH2O) Pperf (cmH2O) 6 6 4 4 D D 2 2 0 0 Acid: + LPS: + + + AngI inj. AngII inj. AngI inj. AngII inj. Supplementary Figure 8

  9. 0.5 0.4 0.3 Fractional shortening acid 0.2 WT, acid 0.1 ace2 KO, acid 0 hrs 0 1 2 3 80 60 (mmHg) 40 Mean arterial pressure acid WT, acid 20 agtr1a-/-, acid 0 0 1 2 3 hrs a b 80 60 (mmHg) Mean arterial pressure 40 acid 20 WT, acid ace2 KO, acid 0 hrs 0 1 2 3 c Supplementary Figure 9

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