1. Fig. 2. DCA activates PDH and increases respiration in human PAH EVLP.
DCA activates PDH and increases respiration in human PAH EVLP. (A) Photograph and schematic of the custom-made EVLP system with a human lung (see Materials and Methods). ICU, intensive care unit. (B to C) PDH activity measured from lung biopsies before and after 1 hour of DCA perfusion (0.7 mg/ml). (B) PDH activity dipstick assay, measuring NADH production from the PDH reaction (see box on top), quantitation [AU (arbitrary units)], and SNP scores. (C) Immunoblots for p-PDH-E1α and quantitation [(p-PDH-E1α/actin)/(total PDH-E1α/actin)]. Lungs 1, 2, and 3 showed an increase in PDH activity by DCA, whereas lung 4 was resistant to DCA (for patient information, see Table 1). The control lung 5 (see Table 1) was perfused with vehicle (veh.) (perfusate) for the same time as lungs 1 to 4. (D) Mitochondrial respiration (oxygen consumption rate) measured using a Seahorse protocol from pre- and post-perfusion biopsies. Top: An example of oxygen consumption rate (slope of each line) of a pre- and post-DCA perfusion biopsy from lung 3. Bottom: The % change in oxygen consumption rate after DCA for all lungs. Perfusion of lung 5 with vehicle (perfusate) was a time control experiment.
Evangelos D. Michelakis et al., Sci Transl Med 2017;9:eaao4583
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