Thromboxane-Induced Contractile Response of Human Coronary Arterioles Is Diminished After Cardioplegic Arrest Jun Feng, MD, PhD, Yuhong Liu, MD, Louis.

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Thromboxane-Induced Contractile Response of Human Coronary Arterioles Is Diminished After Cardioplegic Arrest Jun Feng, MD, PhD, Yuhong Liu, MD, Louis M. Chu, MD, Richard T. Clements, PhD, Kamal R. Khabbaz, MD, Michael P. Robich, MD, Cesario Bianchi, MD, PhD, Frank W. Sellke, MD The Annals of Thoracic Surgery Volume 92, Issue 3, Pages 829-836 (September 2011) DOI: 10.1016/j.athoracsur.2011.04.049 Copyright © 2011 The Society of Thoracic Surgeons Terms and Conditions

Fig 1 (A) Coronary arteriolar vasoconstriction in response to the thromboxane A (TXA)-2 analog U-46619 before vs after cardioplegic ischemia, followed by reperfusion (pre-CP/Rep [white circles] vs post-CP/Rep [black circles]); *p < 0.05 vs pre-CP/Rep. (B) Administration of U-46619 with or without TXA-2 antagonist SQ-29548 to microvessels of pre-CP/Rep (U-46619 [pre-CP/Rep, white circles] or SQ-29548 + U-46619 [pre-CP/Rep, black squares]); *p < 0.05 vs U-46619 (pre-CP/Rep). (C) Infusion of U-46619, with or without the TXA-2 antagonist SQ-29548, to post-CP/Rep vessels (U-46619 [post-CP/Rep, black circles] or SQ-29548 + U-46619 [post-CP/Rep, black diamonds]); p < 0.05 vs U-46619 (post-CP/Rep); n = 6 to 8/group. The bars show the standard error of the mean. The Annals of Thoracic Surgery 2011 92, 829-836DOI: (10.1016/j.athoracsur.2011.04.049) Copyright © 2011 The Society of Thoracic Surgeons Terms and Conditions

Fig 2 (A) Administration of U-46619 to coronary arteriole before cardioplegic ischemia, followed by reperfusion (pre-CP/Rep), with or without the presence of the phospholipase-C (PLC) inhibitor U-73122 (U-46619 [Pre-CP/Rep, white circles] or U-73122 + U-46619 [pre-CP/Rep, white triangles]), *p < 0.05 vs U-46619 (pre-CP/Rep). (B) Perfusion of U-46619 to post-CP/Rep coronary arterioles, with or without the presence of the PLC inhibitor U-73122 (U-46619 [post-CP/Rep, black circles] or U-73122 + U-46619 [post-CP/Rep, black squares]),*p < 0.05 vs. U-46619 (post-CP/Rep); n = 6 to 8/group. The bars show the standard error of the mean. The Annals of Thoracic Surgery 2011 92, 829-836DOI: (10.1016/j.athoracsur.2011.04.049) Copyright © 2011 The Society of Thoracic Surgeons Terms and Conditions

Fig 3 (A) Addition of U-46619 to the vessels from samples before cardioplegic ischemia, followed by reperfusion (pre-CP/Rep), in the absence or presence of the protein kinase C (PKC) inhibitor safingol (U-46619 [pre-CP/Rep, white circles] or safingol + U-46619 [pre-CP/Rep, white diamonds]). (B) Addition of U-46619 to the vessels from post-CP/Rep samples in the absence or presence of PKC inhibitor safingol (U-46619 [post-CP/Rep, black circles] or safingol + U-46619 [post-CP/Rep, black diamonds]); n= 6 to 8/group. The bars show the standard error of the mean. The Annals of Thoracic Surgery 2011 92, 829-836DOI: (10.1016/j.athoracsur.2011.04.049) Copyright © 2011 The Society of Thoracic Surgeons Terms and Conditions

Fig 4 Representative immunoblot of human atria tissue. Lanes 1 to 12, loaded with 40 μg protein, were developed for (A, top) thromboxane A (TXA)-2 receptor and TXA-2 synthase (n = 6 ) and (B, top) phospholipase-C (PLC)-β3, phospho–PLC-β3, PLC-γ1, and phosphor–PLC-γ1 polypeptides (n = 6). Immunoblot band intensity shows unaltered levels of TXA-2 receptor, TXA-2 synthase, PLC-γ1, and PLC-β3 polypeptides after cardioplegic ischemia, followed by reperfusion (CP/Rep; bottom of Fig 4A and B), respectively. The bars show the standard error of the mean. The Annals of Thoracic Surgery 2011 92, 829-836DOI: (10.1016/j.athoracsur.2011.04.049) Copyright © 2011 The Society of Thoracic Surgeons Terms and Conditions

Fig 5 Immunolocalization of thromboxane A (TXA)-2 receptor, phospholipase-C (PLC)-γ1, and PLC-β3 polypeptides in human coronary microvessels (n = 6). Vessels were costained for smooth muscle α-actin with (A) TXA-2 receptor antibody or (B) PLC-β3. Matched negative controls are displayed below each row of primary antibody. (DAPI = 4',6-diamidino-2-phenylindole.) The Annals of Thoracic Surgery 2011 92, 829-836DOI: (10.1016/j.athoracsur.2011.04.049) Copyright © 2011 The Society of Thoracic Surgeons Terms and Conditions