[进展翻译] Circulation 2007年9月11日

1. Increased Endoplasmic Reticulum Stress in Atherosclerotic Plaques Associated With Acute Coronary Syndrome
Masafumi Myoishi, MD; Hiroyuki Hao, MD, PhD; Tetsuo Minamino, MD, PhD; Kouki Watanabe, MD, PhD; Kensaku Nishihira, MD, PhD; Kinta Hatakeyama, MD, PhD; Yujiro Asada, MD, PhD; Ken-ichiro Okada, MD, PhD; Hatsue Ishibashi-Ueda, MD, PhD; Giulio Gabbiani, MD, PhD; Marie-Luce Bochaton-Piallat, PhD; Naoki Mochizuki, MD, PhD; Masafumi Kitakaze, MD, PhD
From the Departments of Cardiovascular Medicine (M.M., M.K.), Structural Analysis (M.M., N.M.), and Pathology (H.H., H.I.-U.), National Cardiovascular Center, Suita, Osaka, Japan; Department of Surgical Pathology (H.H.), Hyogo College of Medicine, Nishinomiya, Hyogo, Japan; Departments of Bioregulatory Medicine (M.M.) and Cardiovascular Medicine (T.M., K.-i.O.), Osaka University Graduate School of Medicine, Suita, Osaka, Japan; Division of Cardiology (K.W.), Uwajima City Hospital, Uwajima, Ehime, Japan; Department of Pathology (K.N., K.H., Y.A.), Faculty of Medicine, University of Miyazaki, Miyazaki, Japan; and Department of Pathology and Immunology (G.G., M.-L.B.-P.), University of Geneva–CMU, Geneva, Switzerland.
Correspondence to Masafumi Kitakaze, MD, PhD, Department of Cardiovascular Medicine, National Cardiovascular Center, Suita, Osaka 565-8565, Japan. E-mail kitakaze@zf6.so-net.ne.jp
Received December 10, 2006; accepted July 6, 2007.
Background— The endoplasmic reticulum (ER) responds to various stresses by upregulation of ER chaperones, but prolonged ER stress eventually causes apoptosis. Although apoptosis is considered to be essential for the progression and rupture of atherosclerotic plaques, the influence of ER stress and apoptosis on rupture of unstable coronary plaques remains unclear.
Methods and Results— Coronary artery segments were obtained at autopsy from 71 patients, and atherectomy specimens were obtained from 40 patients. Smooth muscle cells and macrophages in the fibrous caps of thin-cap atheroma and ruptured plaques, but not in the fibrous caps of thick-cap atheroma and fibrous plaques, showed a marked increase of ER chaperone expression and apoptotic cells. ER chaperones also showed higher expression in atherectomy specimens from patients with unstable angina pectoris than in specimens from those with stable angina. Expression of 7-ketocholesterol was increased in the fibrous caps of thin-cap atheroma compared with thick-cap atheroma. Treatment of cultured coronary artery smooth muscle cells or THP-1 cells with 7-ketocholesterol induced upregulation of ER chaperones and apoptosis, whereas these changes were prevented by antioxidants. We also investigated possible signaling pathways for ER-initiated apoptosis and found that the CHOP (a transcription factor induced by ER stress)-dependent pathway was activated in unstable plaques. In addition, knockdown of CHOP expression by small interfering RNA decreased ER stress-dependent death of cultured coronary artery smooth muscle cells and THP-1 cells.
Conclusions— Increased ER stress occurs in unstable plaques. Our findings suggest that ER stress-induced apoptosis of smooth muscle cells and macrophages may contribute to plaque vulnerability.
2. Visceral and Subcutaneous Adipose Tissue Volumes Are Cross-Sectionally Related to Markers of Inflammation and Oxidative Stress
The Framingham Heart Study
Karla M. Pou, MD; Joseph M. Massaro, PhD; Udo Hoffmann, MD, MPH; Ramachandran S. Vasan, MD; Pal Maurovich-Horvat, MD; Martin G. Larson, ScD; John F. Keaney, Jr, MD, PhD; James B. Meigs, MD, MPH; Izabella Lipinska, PhD; Sekar Kathiresan, MD; Joanne M. Murabito, MD, ScM; Christopher J. O’Donnell, MD, MPH; Emelia J. Benjamin, MD, ScM*; Caroline S. Fox, MD, MPH*
From the National Heart, Lung, and Blood Institute’s Framingham Heart Study, Framingham, Mass (J.M. Massaro, R.S.V., M.G.L., J.M. Murabito, C.J.D., E.J.B., C.S.F.); Division of Endocrinology, Metabolism, and Diabetes, Brigham and Women’s Hospital and Harvard Medical School, Boston, Mass (K.M.P., C.S.F.); Boston University School of Public Health, Departments of Biostatistics (J.M. Massaro, M.G.L.) and Epidemiology (E.J.B.), Boston, Mass; Cardiac MR PET CT Program, Massachusetts General Hospital, Harvard Medical School, Boston, Mass (U.H.); Department of Cardiology (R.S.V., J.F.K., E.J.B.) and Preventive Medicine (R.S.V., E.J.B.), Whitaker Cardiovascular Institute (R.S.V., J.F.K., I.L., E.J.B.), Boston University School of Medicine, Boston, Mass; Semmelweis University, Budapest, Hungary (P.M.-H.); General Medicine Division, Massachusetts General Hospital, Harvard Medical School, Boston, Mass (J.B.M.); the Division of Cardiology, Massachusetts General Hospital and Harvard Medical School, Boston, Mass (C.J.D., S.K.); and Section of General Internal Medicine, Boston University School of Medicine, Boston, Mass (J.M. Murabito).
Correspondence to Caroline S. Fox, MD, MPH, 73 Mt Wayte Ave, Suite 2, Framingham, MA 01702. E-mail foxca@nhlbi.nih.gov
Received April 19, 2007; accepted July 18, 2007.
Background— Excess adiposity is associated with greater systemic inflammation. Whether visceral adiposity is more proinflammatory than subcutaneous abdominal adiposity is unclear.
Methods and Results— We examined the relations of abdominal subcutaneous adipose tissue (SAT) and visceral adipose tissue (VAT), assessed by multidetector computerized tomography, to circulating inflammatory and oxidative stress biomarkers in 1250 Framingham Heart Study participants (52% women; age 60±9 years). Biomarkers were examined in relation to increments of SAT and VAT after adjustment for age, sex, smoking, physical activity, menopause, hormone replacement therapy, alcohol, and aspirin use; additional models included body mass index and waist circumference. SAT and VAT were positively and similarly (with respect to strength of association) related to C-reactive protein, fibrinogen, intercellular adhesion molecule-1, interleukin-6, P-selectin, and tumor necrosis factor receptor-2 (multivariable model R2 0.06 to 0.28 [SAT] and 0.07 to 0.29 [VAT]). However, compared with SAT, VAT was more highly associated with urinary isoprostanes and monocyte chemoattractant protein-1 (SAT versus VAT comparison: isoprostanes, R2 0.07 versus 0.10, P=0.002; monocyte chemoattractant protein-1, R2 0.07 versus 0.08, P=0.04). When body mass index and waist circumference were added to the models, VAT remained significantly associated with only C-reactive protein (P=0.0003 for women; P=0.006 for men), interleukin-6 (P=0.01), isoprostanes (P=0.0002), and monocyte chemoattractant protein-1 (P=0.008); SAT only remained associated with fibrinogen (P=0.01).
Conclusions— The present cross-sectional data support an association between both SAT and VAT with inflammation and oxidative stress. The data suggest that the contribution of visceral fat to inflammation may not be completely accounted for by clinical measures of obesity (body mass index and waist circumference).
3. Prognostic Value of Very Low Plasma Concentrations of Troponin T in Patients With Stable Chronic Heart Failure
Roberto Latini, MD; Serge Masson, PhD; Inder S. Anand, MD; Emil Missov, MD; Marjorie Carlson, BS; Tarcisio Vago, BiolD; Laura Angelici, MS; Simona Barlera, MS; Giovanni Parrinello, PhD; Aldo P. Maggioni, MD; Gianni Tognoni, MD; Jay N. Cohn, MD, for the Val-HeFT Investigators
From the Department of Cardiovascular Research, Istituto di Ricerche Farmacologiche Mario Negri, Milan, Italy (R.L., S.M., L.A., S.B.); Department of Medicine, New York Medical College, Valhalla, NY (R.L.); Cardiology Section, Veterans Affairs Medical Center, Minneapolis, Minn (I.S.A.); Cardiovascular Division, Department of Medicine, University of Minnesota Medical School, Minneapolis (E.M., M.C., J.N.C.); Endocrinology Laboratory, Ospedale Luigi Sacco, Milan, Italy (T.V.); Section of Medical Statistics, University of Brescia, Brescia, Italy (G.P.); Centro Studi ANMCO, Florence, Italy (A.P.M.); and Consorzio Mario Negri Sud, S. Maria Imbaro, Italy (G.T.).
Correspondence to Roberto Latini, MD, Istituto Mario Negri, via La Masa 19, 20156 Milano, Italy. E-mail latini@marionegri.it
Received August 1, 2006; accepted June 11, 2007.
Background— Circulating cardiac troponin T, a marker of cardiomyocyte injury, predicts adverse outcome in patients with heart failure (HF) but is detectable in only a small fraction of those with chronic stable HF. We assessed the prognostic value of circulating cardiac troponin T in patients with stable chronic HF with a traditional (cTnT) and a new precommercial highly sensitive assay (hsTnT).
Methods and Results— Plasma troponin T was measured in 4053 patients with chronic HF enrolled in the Valsartan Heart Failure Trial (Val-HeFT). Troponin T was detectable in 10.4% of the population with the cTnT assay (detection limit 0.01 ng/mL) compared with 92.0% with the new hsTnT assay ( 0.001 ng/mL). Patients with cTnT elevation or with hsTnT above the median (0.012 ng/mL) had more severe HF and worse outcome. In Cox proportional hazards models adjusting for clinical risk factors, cTnT was associated with death (780 events; hazard ratio=2.08; 95% confidence interval, 1.72 to 2.52; P<0.0001) and first hospitalization for HF (655 events; hazard ratio=1.55; 95% confidence interval, 1.25 to 1.93; P<0.0001). HsTnT was associated with the risk of death in unadjusted analysis for deciles of concentrations and in multivariable models (hazard ratio=1.05; 95% confidence interval, 1.04 to 1.07 for increments of 0.01 ng/mL; P<0.0001). Addition of hsTnT to well-calibrated models adjusted for clinical risk factors, with or without brain natriuretic peptide, significantly improved prognostic discrimination (C-index, P<0.0001 for both outcomes).
Conclusions— In this large population of patients with HF, detectable cTnT predicts adverse outcomes in chronic HF. By the highly sensitive assay, troponin T retains a prognostic value at previously undetectable concentrations.
4. In Vivo Characterization of Murine Myocardial Perfusion With Myocardial Contrast Echocardiography
Validation and Application in Nitric Oxide Synthase 3–Deficient Mice
Michael J. Raher, BS*; Hélène Thibault, MD*; Kian Keong Poh, MD; Rong Liu, MD, PhD; Elkan F. Halpern, PhD; Geneviève Derumeaux, MD, PhD; Fumito Ichinose, MD; Warren M. Zapol, MD; Kenneth D. Bloch, MD; Michael H. Picard, MD; Marielle Scherrer-Crosbie, MD, PhD
From the Cardiac Ultrasound Laboratory in the Cardiology Division of the Department of Medicine (M.J.R., H.T., K.K.P., M.H.P., M.S.-C.), the Cardiovascular Research Center (M.J.R., H.T., K.D.B., M.S.-C.), the Department of Anesthesia and Critical Care (M.J.R., R.L., F.I., W.M.Z., K.D.B.), and the Institute for Technology Assessment (E.F.H.), Massachusetts General Hospital and Harvard Medical School, Boston, Mass; and INSERM E 0226 (H.T., G.D.), Université Claude Bernard Lyon I, Lyon, France.
Correspondence to Marielle Scherrer-Crosbie, MD, PhD, Cardiac Ultrasound Laboratory, Massachusetts General Hospital, 55 Fruit St, Boston, MA 02114. E-mail marielle@crosbie.com
Received November 4, 2006; accepted June 29, 2007.
Background— The ability to noninvasively evaluate murine myocardial blood flow (MBF) in vivo would provide an important tool for cardiovascular research. Myocardial contrast echocardiography (MCE) has been used to measure MBF; however, it has not been validated in mice. This study assesses whether MCE can evaluate MBF at rest and after vasodilation and measure the maximal augmentation (coronary reserve) of MBF in mice. Wild-type (WT) and nitric oxide synthase 3 (NOS3)–deficient (NOS3–/–) mice were studied.
Methods and Results— MCE was performed at baseline and after intravenous infusion of acetylcholine or adenosine. Definity contrast agent was infused, and parasternal views were acquired in real-time mode. Replenishment curves of myocardial contrast were obtained, and rates of signal rise (ß) and plateau intensity (A) were calculated. MBF estimated by the product of A and ß (Aß) was compared with that measured with fluorescent microspheres. MCE analysis was feasible in 98% (52/53) of mice. MBF measured by microspheres increased with adenosine and correlated closely with Aß. There was no difference in MCE-derived MBF between WT and NOS3–/– mice at rest. Adenosine infusion increased MBF by 3.0±0.6-fold in NOS3–/– mice and 2.5±0.3-fold in WT (P=0.58 between genotypes). Acetylcholine induced an increase of 2.4±0.2-fold in MBF in WT mice but did not increase MBF in NOS3–/– mice (P<0.0005 versus WT).
Conclusions— MBF, coronary reserve, and vasodilator responses can be evaluated accurately in the intact mouse by MCE. This method demonstrated a preserved coronary response to adenosine but an impaired acetylcholine-induced vasodilation in NOS3–/– mice compared with WT mice.
5. Bone Morphogenic Protein Antagonists Are Coexpressed With Bone Morphogenic Protein 4 in Endothelial Cells Exposed to Unstable Flow In Vitro in Mouse Aortas and in Human Coronary Arteries
Role of Bone Morphogenic Protein Antagonists in Inflammation and Atherosclerosis
Kyunghwa Chang, PhD; Daiana Weiss, MD; Jin Suo, PhD; J. David Vega, MD; Don Giddens, PhD; W. Robert Taylor, MD, PhD; Hanjoong Jo, PhD
From the Wallace H. Coulter Department of Biomedical Engineering (K.C., J.S., D.G., W.R.T., H.J.), Georgia Institute of Technology and Emory University, Atlanta, Ga, and the Department of Surgery (J.D.V.), and Division of Cardiology (D.W., W.R.T., H.J.), Emory University, Atlanta, Ga.
Correspondence to Hanjoong Jo, PhD, Wallace H. Coulter Department of Biomedical Engineering at Georgia Tech and Emory University, 2005 WMB, Atlanta, GA 30322. E-mail hanjoong.jo@bme.gatech.edu
Received December 11, 2006; accepted June 29, 2007.
Background— Exposure to disturbed flow, including oscillatory shear stress, stimulates endothelial cells (ECs) to produce bone morphogenic protein (BMP) 4, which in turn activates inflammation, a critical atherogenic step. BMP activity is regulated by the level of BMP antagonists. Until now it was not known whether shear also regulates the expression of BMP antagonists and whether they play a role in EC pathophysiology.
Methods and Results— BMP antagonists follistatin, noggin, and matrix Gla protein were expressed in cultured bovine and human arterial ECs. Surprisingly, oscillatory shear stress increased expression of the BMP antagonists in ECs, whereas unidirectional laminar shear decreased such expression. Immunohistochemical studies with mouse aortas showed data consistent with in vitro findings: Only ECs in the lesser curvature exposed to disturbed flow, but not those in the greater curvature and straight arterial regions exposed to undisturbed flow, showed coexpression of BMP4 and the BMP antagonists. Similarly, in human coronary arteries, expression of BMP4 and BMP antagonists in ECs positively correlated with the severity of atherosclerosis. Monocyte adhesion induced by oscillatory shear stress was inhibited by knockdown of BMP4 or treatment with recombinant follistatin or noggin, whereas it was increased by knockdown of follistatin and/or noggin.
Conclusions— The present results suggest that ECs coexpress BMP antagonists along with BMP4 in an attempt to minimize the inflammatory response by oscillatory shear stress as part of a negative feedback mechanism. The balance between the agonist, BMP4, and its antagonists may play an important role in the overall control of inflammation and atherosclerosis.
6. Expression of Cholesteryl Ester Transfer Protein in Mice Promotes Macrophage Reverse Cholesterol Transport
Hiroyuki Tanigawa, MD, PhD; Jeffrey T. Billheimer, PhD; Jun-ichiro Tohyama, MD; YuZhen Zhang, MD, PhD; George Rothblat, PhD; Daniel J. Rader, MD
From the Institute for Translational Medicine and Therapeutics and the Cardiovascular Institute (H.T., J.T.B., J.-i.T., Y.Z.Z., D.J.R.), University of Pennsylvania School of Medicine, Philadelphia, and the Children’s Hospital of Philadelphia (G.R.), Pa.
Correspondence to Daniel J. Rader, MD, University of Pennsylvania Medical Center, 654 Biomedical Research Building II/III, 421 Curie Blvd, Philadelphia, PA 19104. E-mail rader@mail.med.upenn.edu
Received March 21, 2007; accepted June 29, 2007.
Background— Cholesteryl ester transfer protein (CETP) transfers cholesteryl esters from high-density lipoproteins to apolipoprotein (apo) B–containing lipoproteins and in humans plays an important role in lipoprotein metabolism. However, the role that CETP plays in mediation of reverse cholesterol transport (RCT) remains unclear. We used a validated in vivo assay of macrophage RCT to test the effect of CETP expression in mice (which naturally lack CETP) on macrophage RCT, including in mice that lack the low-density lipoprotein receptor or the scavenger receptor class B, type I.
Method and Results— A vector based on adeno-associated virus serotype 8 (AAV8) with a liver-specific thyroglobulin promoter was used to stably express human CETP in livers of mice and was compared with an AAV8-lacZ control vector. The RCT assay was performed 4 weeks after vector injection and involved the intraperitoneal injection of acetylated low-density lipoprotein cholesterol–loaded and 3H-cholesterol–labeled J774 macrophages in mice with plasma sampling at several time points, liver and bile sampling at 48 hours, and continuous fecal collection to measure 3H-sterol as an integrated readout of macrophage RCT. In apobec-1–null mice, CETP expression reduced plasma high-density lipoprotein cholesterol levels but significantly increased fecal 3H-sterol excretion. In low-density lipoprotein receptor/apobec-1 double-null mice, CETP expression reduced high-density lipoprotein cholesterol levels and had no effect on fecal 3H-sterol excretion. Finally, in scavenger receptor class B, type I–null mice, CETP expression reduced high-density lipoprotein cholesterol levels and significantly increased fecal 3H-sterol excretion.
Conclusion— The present results demonstrate that CETP expression promotes macrophage RCT in mice, that this effect is dependent on the low-density lipoprotein receptor, and that CETP expression restores to normal the impaired RCT in mice deficient in scavenger receptor class B, type I.
7. Prognostic Impact of Microvasculopathy on Survival After Heart Transplantation
Evidence From 9713 Endomyocardial Biopsies
Nicola E. Hiemann, MD; Ernst Wellnhofer, MD; Christoph Knosalla, MD, PhD; Hans B. Lehmkuhl, MD; Julia Stein, MSc; Roland Hetzer, MD, PhD; Rudolf Meyer, MD, PhD
From the Departments of Cardiothoracic and Vascular Surgery (N.E.H., C.K., H.B.L., J.S., R.H., R.M.) and Cardiology (E.W.), Deutsches Herzzentrum Berlin, Berlin, Germany.
Reprint requests to Nicola E. Hiemann, MD, Department of Cardiothoracic and Vascular Surgery, Deutsches Herzzentrum Berlin, Augustenburger Platz 1, 13353 Berlin, Germany. E-mail hiemann@dhzb.de
Received June 20, 2006; accepted July 3, 2007.
Background— Epicardial vasculopathy has been shown to be associated with poor outcome after heart transplantation. We demonstrate that histologically proven stenotic microvasculopathy is a novel prognostic factor for long-term survival.
Methods and Results— In 9713 biopsies harvested within the first posttransplantation year from 873 patients (83% male; mean age, 49.1±0.6 years), light microscopic evaluations (x200) were performed for microvasculopathy, defined as stenotic endothelial and/or medial disease. Prevalence of severe epicardial vasculopathy was defined by presence of 75% luminal stenosis in coronary angiography (available in 611 of 873 patients), which was present in 118 of 611 patients (19%). For Kaplan-Meier analysis, we defined fatal cardiac events as lethal acute myocardial infarction, sudden cardiac death, and graft failure. Stenotic microvasculopathy was present in 379 of 873 patients (43%) and was due to medial (345/379; 91%) rather than endothelial disease (2/379; 1%) or a combination of both (31/379; 8%; P<0.001). Endothelial disease (median [95% CI], 12.07 [10.69 to 13.44] versus 12.73 years [10.16 to 15.30]; P=0.3329) and nonstenotic medial disease (12.44 [11.14 to 13.74] versus 12.43 years [10.51 to 14.35]; P=0.4047) did not decrease overall survival or time to fatal cardiac event. Stenotic microvasculopathy was associated with poor overall survival (10.90 [9.16 to 12.60] versus 13.40 years [11.79 to 15.07]; P=0.0374) and decreased freedom from fatal cardiac events (1, 5, 10 years, 95.6±1.4%, 86.9±2.3%, 75.5±3.1% versus 99.1±0.5%, 96.8±1.0%, 89.8±1.9%; P<0.0001). This finding was independent of epicardial transplant vasculopathy (P=0.0031).
Conclusions— Stenotic microvasculopathy is frequent in routinely processed biopsies and a new prognostic factor for long-term survival after heart transplantation.
8. Distinct Downregulation of C-Type Natriuretic Peptide System in Human Aortic Valve Stenosis
Tuomas O. Peltonen, MD; Panu Taskinen, MD, PhD; Ylermi Soini, MD, PhD; Jaana Rysä, MSc; Jarkko Ronkainen, MD; Pasi Ohtonen, MSc; Jari Satta, MD, PhD; Tatu Juvonen, MD, PhD; Heikki Ruskoaho, MD, PhD; Hanna Leskinen, MD, PhD
From the Department of Pharmacology and Toxicology (T.O.P., J. Rysä, H.R., H.L.) and Department of Physiology (J. Ronkainen), Biocenter Oulu, and the Department of Pathology (Y.S.), University of Oulu, Oulu, Finland; and the Department of Cardiovascular Surgery (P.T., J.S., T.J.) and Departments of Anaesthesiology and Surgery (P.O.), Oulu University Hospital, Oulu, Finland.
Correspondence to Heikki Ruskoaho, Department of Pharmacology and Toxicology, University of Oulu, PO Box 5000, 90014 Oulu, Finland. E-mail heikki.ruskoaho@oulu.fi
Received December 22, 2006; accepted July 9, 2007.
Background— Aortic valve calcification is an actively regulated process that displays hallmarks of atherosclerosis. Natriuretic peptides (A-, B-, and C-type natriuretic peptides [ANP, BNP, and CNP]) have been reported to have a role in the pathogenesis of vascular atherosclerosis, but their expression in aortic valves is not known. Here, we characterized and compared expression of natriuretic peptide system in aortic valves of patients with normal valves (n=4), aortic regurgitation (n=11), regurgitation and fibrosis (n=6), and aortic valve stenosis (n=21).
Methods and Results— By reverse-transcription polymerase chain reaction, all 3 natriuretic peptides were found to be expressed in aortic valves. CNP mRNA levels were 92% lower (P<0.001) in stenotic valves, whereas no significant changes in the expression of ANP and BNP genes were found compared with valves obtained from patients with aortic regurgitation. CNP was localized by immunohistochemistry with specific CNP (32-53) antibody to valvular endothelial cells and myofibroblasts. Gene expression of furin, which proteolytically cleaves proCNP into active CNP, was 54% lower in aortic valve stenosis (P=0.04). Moreover, natriuretic peptide receptor-A and natriuretic peptide receptor-B mRNA levels were 78% and 76% lower, respectively, in stenotic valves. In contrast, gene expression of corin, a proANP- and proBNP-converting enzyme, and natriuretic peptide receptor-C did not differ between groups.
Conclusions— We show that natriuretic peptides, their processing enzymes, and their receptors are expressed in human aortic valves. Aortic valve stenosis is characterized by distinct downregulation of gene expression of CNP, its processing enzyme furin, and the target receptors natriuretic peptide receptor-B and natriuretic peptide receptor-A, which suggests that CNP acts as a paracrine regulator of the aortic valve calcification process.