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Diabetes Upregulation of Cyclooxygenase 2 Contributes to Altered Coronary Reactivity After Cardiac Surgery  Jun Feng, MD, PhD, Kelsey Anderson, Arun K.

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Presentation on theme: "Diabetes Upregulation of Cyclooxygenase 2 Contributes to Altered Coronary Reactivity After Cardiac Surgery  Jun Feng, MD, PhD, Kelsey Anderson, Arun K."— Presentation transcript:

1 Diabetes Upregulation of Cyclooxygenase 2 Contributes to Altered Coronary Reactivity After Cardiac Surgery  Jun Feng, MD, PhD, Kelsey Anderson, Arun K. Singh, MD, Afshin Ehsan, MD, Hunter Mitchell, BS, Yuhong Liu, MD, Frank W. Sellke, MD  The Annals of Thoracic Surgery  Volume 104, Issue 2, Pages (August 2017) DOI: /j.athoracsur Copyright © 2017 The Society of Thoracic Surgeons Terms and Conditions

2 Fig 1 (A) The relaxation responses of nondiabetes mellitus (ND [circles]), controlled diabetes (CDM [squares]), and uncontrolled diabetes (UDM [triangles]) coronary arterioles to the dose-dependent, endothelium-dependent vasodilator bradykinin at baseline, before cardioplegia arrest and cardiopulmonary bypass (pre-CP/CPB). (B) The relaxation response of ND, CDM, and UDM coronary arterioles to bradykinin after CP/CPB (post-CP/CPB). (C) The relaxation response of ND coronary arterioles to bradykinin between pre-CP/CPB (circles) and post-CP/CPB (squares). (D) The relaxation response of CDM coronary arterioles to bradykinin between pre-CP/CPB and post-CP/CPB. (E) The relaxation response of UDM coronary arterioles to bradykinin between pre-CP/CPB and post-CP/CPB. Data are mean ± standard error of the mean; n = 12 per group. *p < 0.05 versus ND or CDM or pre-CP/CPB. **p < versus pre-CP/CPB. The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur ) Copyright © 2017 The Society of Thoracic Surgeons Terms and Conditions

3 Fig 2 Relaxation response of arterioles to bradykinin before cardioplegia arrest and cardiopulmonary bypass (pre-CP/CPB) with (squares) and without (circles) the selective cyclooxygenase (COX)-2 inhibitor NS398 from (A) no diabetes mellitus (ND) patients, (C) controlled diabetes (CDM) patients, and (E) uncontrolled diabetes (UDM) patients. The relaxation response of post-CP/CPB arterioles to bradykinin with (squares) and without (circles) the selective COX-2 inhibitor NS398 from (B) ND, (D) CDM, and (F) UDM patients. Data are mean ± standard error of the mean; n = 6 per group. *p < 0.05 versus bradykinin alone. **p < versus bradykinin alone. The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur ) Copyright © 2017 The Society of Thoracic Surgeons Terms and Conditions

4 Fig 3 (A, C, E) Representative immunoblots of human atrial tissue samples from no diabetes mellitus (ND) patients, controlled diabetes (CDM) patients, and uncontrolled diabetes (UDM) patients. Lanes 1 to 6 loaded with 40 μg protein were developed for cyclooxygenase (COX)-1, COX-2, and nicotinamide adenine dinucleotide phosphate oxidase (NOX)-1 polypeptides. (GAPDH = glyceraldehyde 3-phosphate dehydrogenase.) (B) Densitometric evaluation of immunoblot band intensity shows no significant differences in the levels of COX-1 among three groups, or between pre-cardioplegia arrest and cardiopulmonary bypass (pre-CP/CPB [black bars]) and post-CP/CPB (gray bars). (D) Densitometric evaluation of immunoblot band intensity shows significant differences in the levels of COX-2 between UDM and ND or CDM groups, and between pre- and post-CP/CPB. (F) Densitometric evaluation of immunoblot band intensity shows significant differences in the levels of NOX-1 between UDM and ND or CDM groups, or between before and after CP/CPB. Data are mean ± standard error of the mean; n = 6 per group. *p < 0.05 versus < 0.05 versus ND pre-CP/CPB or CDM pre-CP/CPB. #p < 0.05 versus ND post-CP/CPB or CDM post-CP/CPB. (O.D. = optical density.) The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur ) Copyright © 2017 The Society of Thoracic Surgeons Terms and Conditions

5 Fig 4 (A, B) Immunohistochemical image of COX-1 and COX-2 in paraffin-embedded human atrial tissues from no diabetes mellitus (ND) and diabetes mellitus (DM) patients. (CP/CPB = cardioplegia arrest and cardiopulmonary bypass.) (C) Densitometric analysis of signal intensities shows no significant differences in cyclooxygenase (COX)-1 distribution among the three groups or between pre-CP/CPB (black bars) and post-CP/CPB (gray bars). (D) Densitometric analysis of signal intensities shows differences in COX-2 distribution among the three groups or between pre- and post-CP/CPB. (E) Immunofluorescence staining of COX-2. Vessels were costained for cell nuclei (blue), smooth muscle α-actin (green), and COX-2 (red in endothelium and orange in smooth muscle). (F) Densitometric analysis of immunofluorescence intensities shows significant difference in COX-2 distribution in the coronary vessels among the three groups or between pre-CP/CPB (black bars) and post-CP/CPB (gray bars). Data are mean ± standard error of the mean; n = 6 per group. *p < 0.05 versus < 0.05 versus ND pre-CP/CPB or CDM pre-CP/CPB. #p < 0.05 versus ND post-CP/CPB or CDM post-CP/CPB. (CDM = controlled diabetes; O.D. = optical density; UDM = uncontrolled diabetes.) The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur ) Copyright © 2017 The Society of Thoracic Surgeons Terms and Conditions

6 Fig 5 (A) Immunofluorescence staining of nicotinamide adenine dinucleotide phosphate oxidase (NOX)-2 and NOX-4 before cardioplegia arrest and cardiopulmonary bypass (pre-CP/CPB) and post-CP/CPB for no diabetes mellitus (ND), controlled diabetes (CDM), and uncontrolled diabetes (UDM) patients. Vessels were costained for cell nuclei (blue), smooth muscle α-actin (green), and NOX-2 or NOX-4 endothelium (red) and smooth muscle (orange). Bar graphs showing increased distribution of (B) NOX-2 and (C) NOX-4 in diabetes myocardium. Data are mean ± standard error of the mean; n = 6 per group. *p < 0.05 versus < 0.05 versus ND pre-CP/CPB. #p < 0.05 versus ND after CP/CPB or CDM post-CP/CPB. (O.D. = optical density.) The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur ) Copyright © 2017 The Society of Thoracic Surgeons Terms and Conditions

7 Fig 6 (A) Oxyblot protein oxidation detection for the no diabetes mellitus (ND), controlled diabetes (CDM), and uncontrolled diabetes (UDM) groups before (Pre) and after (Post) cardioplegia arrest and cardiopulmonary bypass (CP/CPB). (B) Protein oxidation of atrial tissue samples was significantly increased in the UDM and CDM groups. (Black bars = pre-CP/CPB; gray bars = post-CP/CPB.) Data are mean ± standard error of the mean; n = 6 per < 0.05 versus ND pre-CP/CPB or CDM pre-CP/CPB. #p < 0.05 versus ND post-CP/CPB or CDM post-CP/CPB. (O.D. = optical density.) The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur ) Copyright © 2017 The Society of Thoracic Surgeons Terms and Conditions

8 Fig 7 Proposed mechanisms responsible for diabetes mellitus and cardioplegia arrest and cardiopulmonary bypass (CP/CPB) upregulation of cyclooxygenase (COX)-2. (I/R = ischemia/reperfusion; NOX = nicotinamide adenine dinucleotide phosphate oxidase; PGI = prostacyclin; TXA = thromboxane.) The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur ) Copyright © 2017 The Society of Thoracic Surgeons Terms and Conditions

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