Roland Henaine, MD, Mathieu Vergnat, MD, Emile A

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Presentation transcript:

Effects of lack of pulsatility on pulmonary endothelial function in the Fontan circulation Roland Henaine, MD, Mathieu Vergnat, MD, Emile A. Bacha, MD, Bruno Baudet, BSc, Virginie Lambert, MD, Emre Belli, MD, Alain Serraf, MD, PhD The Journal of Thoracic and Cardiovascular Surgery Volume 146, Issue 3, Pages 522-529 (September 2013) DOI: 10.1016/j.jtcvs.2012.11.031 Copyright © 2013 The American Association for Thoracic Surgery Terms and Conditions

Figure 1 Example of lung sections ipsilateral to A, B, cavopulmonary anastomosis and C, control lung obtained from same pig at 3 months (representative of groups II and III) after cavopulmonary anastomosis, stained with hematoxylin-phloxin-saffron (magnification ×25; scale bar, 500 μm). Arrowheads demonstrate hyperplasia of venous wall under pleura in Figure A, along bronchial axes and between alveolar lobules in Figure B of shunted lung. C, Control lung demonstrating normal histologic findings with normal venous wall thickness. The Journal of Thoracic and Cardiovascular Surgery 2013 146, 522-529DOI: (10.1016/j.jtcvs.2012.11.031) Copyright © 2013 The American Association for Thoracic Surgery Terms and Conditions

Figure 2 Dose-dependent responses of group II (nonpulsatile) right and left pulmonary artery (RPA and LPA, respectively) rings to cumulative addition of A, phenylephrine, B, acetylcholine, and C, calcium ionophore. Dose-dependent responses of group III (micropulsatile) RPA and LPA rings to cumulative addition of D, phenylephrine, E, acetylcholine, and F, calcium ionophore. A, D, Data presented as mean ± standard deviation of developed tension in grams for contraction. B, C, E, F, Data presented as mean ± standard deviation of percentage of dilatation relative to phenylephrine-induced preconstriction level for relaxation. RPA of group II demonstrated significantly different contraction and reduced relaxation (nonsignificant) relative to LPA. A, Phenylephrine-induced (10−9-3.10−4 M) contraction in PA rings. Two-way analysis of variance revealed that RPA rings exhibited greater contraction to phenylephrine than LPA rings (Emax, P = .001; EC50, P = .323). B, Acetylcholine-induced (10−9-10−4 M) relaxation in phenylephrine-preconstricted PA rings. Despite reduced RPA ring relaxation, no significant difference was found (Emax, P = .187; EC50, P = .210). C, Calcium ionophore-induced (10−10-3.10−7 M) relaxation in phenylephrine-preconstricted PA rings. Two-way analysis of variance revealed that RPA rings exhibited blunted relaxation responses to calcium ionophore compared with responses from left lung (Emax, P = .07; EC50, P = .262). RPA of group III demonstrated restored similar level of contraction and relaxation, relative to LPA. D, Phenylephrine-induced (10−9-3.10−4 M) contraction in PA rings. Two-way analysis of variance revealed that restored pulsatility abolished abnormal right pulmonary response with no difference in contraction to phenylephrine between RPA and LPA rings (Emax, P = .715; EC50, P = .08). E, Acetylcholine-induced (10−9-10−4 M) relaxation in phenylephrine-preconstricted PA rings. Two-way analysis of variance revealed that restored pulsatility abolished abnormal right pulmonary response, with no difference in relaxation to acetylcholine between RPA and LPA rings (Emax, P = .495; EC50, P = .233). F, Calcium ionophore-induced (10−10-3.10−7 M) relaxation in phenylephrine-preconstricted PA rings. Two-way analysis of variance revealed that restored pulsatility abolished abnormal right pulmonary response, with no difference in relaxation to calcium ionophore between RPA and LPA rings (Emax, P = .796; EC50, P = .879). The Journal of Thoracic and Cardiovascular Surgery 2013 146, 522-529DOI: (10.1016/j.jtcvs.2012.11.031) Copyright © 2013 The American Association for Thoracic Surgery Terms and Conditions

Figure 3 Dose-dependent responses of group I (sham), group II (nonpulsatile), and group III (micropulsatile) right pulmonary artery (PA) rings to cumulative addition of phenylephrine, acetylcholine, calcium ionophore, and nitroprusside. In right PA, no difference in contraction was found among 3 groups, but significantly reduced dilatation response was found in groups II and III, relative to sham group I. A, Phenylephrine-induced (10−9-3.10−4 M) contraction in PA rings. Two-way analysis of variance revealed contractile responses of right PA rings were similar among groups. B, Acetylcholine-induced (10−9-10−4 M) relaxation in phenylephrine-preconstricted PA rings. Despite reduced relaxation in groups II and III compared with group I, no significant difference was found (groups II and III compared with group I, Emax, P = .333 and P = .334, respectively). C, Calcium ionophore-induced (10−10-3.10−7 M) relaxation in phenylephrine-preconstricted PA rings. Two-way analysis of variance revealed that group II and III right PA rings exhibited blunted relaxation responses to calcium ionophore compared with responses of sham group I (groups II and III compared with group I, Emax, P = .002 and P = .0003, EC50, P = .003 and P = .001, respectively). No significant difference was found in relaxation responses between groups II and III. The Journal of Thoracic and Cardiovascular Surgery 2013 146, 522-529DOI: (10.1016/j.jtcvs.2012.11.031) Copyright © 2013 The American Association for Thoracic Surgery Terms and Conditions

Figure 4 Immunoblot analysis for endothelial nitric oxide synthase (eNOS) protein in lung tissue from 3 groups. Top, Representative immunoblot from protein extracts prepared from lung tissue of pig from indicated group separated on 7.5% sodium dodecyl sulfate-polyacrylamide gel, electrophoretically transferred to Hybond membranes, and analyzed using a specific antiserum raised against eNOS. Bottom, Summary data from 10 pigs/group; densitometric values for eNOS protein normalized to β-actin protein for eNOS protein/β-actin protein. Data presented as mean ± standard deviation, normalized. eNOS protein/β-actin protein decreased by 39% in nonpulsatile (group II, P = .045) and 17% in micropulsatility (group III, P = .39). The Journal of Thoracic and Cardiovascular Surgery 2013 146, 522-529DOI: (10.1016/j.jtcvs.2012.11.031) Copyright © 2013 The American Association for Thoracic Surgery Terms and Conditions