Spontaneous Cholesterol Crystal Embolism in Patients with Chronic Heart Failure

S170 Journal of Cardiac Failure Vol. 18 No. 10S October 2012 Departmet of Cardio-angiology, Kitasato University School of Medicine, Kanagawa, Japan Purpose: The mechanism of cardiac remodeling under high glucose condition still remain unclear even diabetes clearly promoted the arteriosclerosis. Therefore, we analyzed cellular biological function under high glucose condition using the cardiac fibroblast and measured the expression of soluble vascular growth factor receptor -1 (sVEGFR-1) considered to suppress growth factor signals. Materials and Methods: Confluent fibroblasts derived from myocardium of mice were cultured under 0.05, 5, 15 and 30 mM of glucose, respectively and used for migration assay after wound scratching or proliferation assay using MTT assay methods, after 18 hours of glucose addition. H2O2, a major source of reactive oxygen species (ROS) in the cells and sVEGF-R1 in supernatant of the cells were measured by DCF-DA and ELISA, respectively. Results: Resulted migration was significantly increased in 15 mM of glucose (2.6-time increase) but not in another concentration and the proliferation was significantly high over 5 mM (2.3-time increase) compared to 0.05 mM of glucose, respectively. H2O2 was significantly increased over 5 mM of glucose and sVEGF-R1 increased at 5 mM (274.2 +/- 16.6 pg/mL, mean +/- SE) but significantly decreased over 15 mM (182.1 +/- 11.0 pg/mL, P!0.05) Conclusions: Our data suggested that high glucose condition induced the migration and proliferation of fibroblasts, accompanied with ROS and sVEGF-R1 elevation and may contribute to clarify the process of diabetic cardiomyopathy.

P-020 The Overexpression of Mitochondrial Transcription Factor A Attenuates Mitochondrial Reactive Oxygen Species Generation and Inhibits Pathological Remodeling in Cardiac Myocytes TAKEO FUJINO, TOMOMI IDE, MASATAKA IKEDA, KEN ONITSUKA, YUKO HATA, TAKAKO TAKEHARA, KAZUO SAKAMOTO, TOMOYUKI TOBUSHI, KEITA SAKU, KENJI SUNAGAWA Department of Cardiovascular Medicine, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan Background: Mitochondrial transcription factor A (TFAM), a nucleoid protein of mitochondrial DNA (mtDNA), is essential for maintenance of mtDNA. In myocardial infarction, the overexpression of TFAM attenuated the decrease in mtDNA copy number, ameliorated cardiac remodeling and markedly improved survival. However, the mechanism how TFAM attenuates cardiac remodeling remains unknown. Meanwhile, reactive oxygen species (ROS) from mitochondria is associated with the progression of cardiac remodeling. Thus we hypothesized that TFAM attenuates cardiac remodeling via inhibiting mitochondrial ROS. Methods and Results: We prepared adenovirus vector encoding human TFAM. TFAM protein was successfully translocated into mitochondria in cardiac myocytes and increased mtDNA copy number significantly (1.860.2-folds, p!0.01). We stimulated myocytes with angiotensin II (AngII), a major neurohumoral factor in cardiac remodeling. TFAM suppressed AngII-induced mitochondrial ROS generation significantly (p!0.01). To elucidate the effect of TFAM on ROS-related signals in cardiac remodeling, we investigated nuclear factor of activated T cells (NFAT) signaling, a ROS-regulated major transcriptional factor promoting pathological remodeling. TFAM abolished AngII-induced NFAT nuclear translocation, suppressing NFAT transcriptional activity and NFAT-dependent gene expression. Conclusion: The overexpression of TFAM increased mtDNA copy number, and attenuated AngII-induced mitochondrial ROS and subsequent signals of cardiac remodeling in cardiac myocytes. The increase of TFAM could be a potential therapeutic strategy of cardiac remodeling and failure.

P-021 Changes of b-catenin Protein Precede Gap Junction Remodeling in Cardiomyocyte Caused by Rapid Electrical Stimulation TADAMITSU NAKASHIMA, TOMOKO OHKUSA, MASAAKI YOSHIDA, MASUNORI MATSUZAKI Division of Cardiology, Department of Medicine and Clinical Science, Yamaguchi University Graduate School of Medicine, Ube, Japan The intercalated disc (ID) contains two complexes, adhesion junction (AJ), and connexin (Cx) gap junction (GJ) that enable the myocardium to function as a syncytium. The AJ mediates normal mechanical coupling between cardiomyocytes and plays an important role in the formation and stability of GJ. GJs provide the pathway for intercellular current flow, enabling coordinated action potential propagation and contraction. Recently, some studies demonstrated cardiac-specific changes of Cx43 cause abnormal conduction velocity and induce unidirectional block resulting in an arrhythmogenic substrate. We studied the ID remodeling caused by rapid electrical stimulation (RES) in cardiomyocytes using Western blotting and immunohistochemical methods. Neonatal rat ventricular myocytes cultured for 5 days were subjected to RES (field stimulation) at 3.0 Hz. Cx43 expression in cardiomyocytes were significantly increased (1.3 times vs 0 min. p!0.05, n56 each) after 60 min RES. b-catenin expressions in total cell fraction tended to increase during RES up to 60 min. Interestingly, the expression level of b-catenin at nucleus, which functions as T cell factor/ lymphocyte enhancer binding factor (TCF/LEF) transcriptional activator of Cx43, was increased (1.2 times vs 0 min. n54 each) after 10 min RES. These results suggest that changes in AJ protein, b-catenin, precede Cx43 GJ remodeling during RES exposure to cardiomyocytes.

P-022 The Activation of Invariant Natural Killer T Cells Ameliorates Myocardial Ischemia Reperfusion Injury in Mice TSUNEAKI HOMMA, SHINTARO KINUGAWA, ARATA FUKUSHIMA, TADASHI SUGA, SHINGO TAKADA, YOSHIHIRO MASAKI, TOMOYASU KADOGUCHI, TAKAAKI FURIHATA, HIROYUKI TSUTSUI Department of Cardiovascular Medicine, Hokkaido University Graduate School of Medicine, Sapporo, Japan Background: Invariant natural killer T (iNKT) cells play an important role in tissue inflammation in various diseases due to the capacity to produce inflammatory cytokines and orchestrate tissue inflammation. No previous studies have examined the pathophysiological role of iNKT cells in myocardial ischemia/reperfusion (I/R) injury. Methods and Results: I/R or sham operation was performed in male C57BL/6J mice. I/R mice received the injection of either a-galactosylceramide (aGC, I/R+aGC, n514), which specifically activates iNKT cells, or vehicle (I/R+vehicle, n516) 30 minutes before reperfusion. At 24 hours after reperfusion, infarct size was significantly smaller in I/R+aGC than I/R+vehicle (infarct size/area at risk: 37.862.7% vs 47.162.5%, P!0.05), with no significant changes in area at risk. Serum Troponin-I was also lowered in I/R+aGC. The numbers of infiltrating myeloperoxidase and CD3 positive cells were lower in I/R+aGC. In parallel with the decrease of inflammatory cell infiltration, myocardial mRNA expression of tumor necrosis factor-a (TNF-a) was lower in I/R+aGC compared to I/R+vehicle. On the other hand, interleukin-10 (IL-10) mRNA expression was significantly increased after aGC injection. The ratio of mRNA expression of IL-10 to TNF-a was significantly higher in I/R+aGC compared to I/R+vehicle. Anti-IL-10 receptor antibody abrogated these protective effects of aGC on I/R injury. Conclusions: The activation of iNKT cells ameliorated myocardial ischemia reperfusion injury in mice possibly through the enhanced expression of IL-10.

P-023 Spontaneous Cholesterol Crystal Embolism in Patients with Chronic Heart Failure YU KATATA, TSUYOSHI OGATA, HIROSHI FUJITA, NOBUHIRO YAMAGUCHI, NORIKO ONOUE, MITSUAKI TANAKA, TAKESHI ISHIZUKA, TSUYOSHI SHINOZAKI Department of Cardiovascular Medicine, National Hospital Organization Sendai Medical Center, Sendai, Japan Background: Since vascular manipulation is a major cause of cholesterol crystal embolism (CCE), little attention has been directed to spontaneous CCE. Also relationship between heart failure and spontaneous CCE remains unclear. We investigated characteristics of patients with spontaneous CCE. Results: CCE occurred in 6 patients out of 66441 patients hospitalizing from April 2006 to March 2011 in our hospital. In 3 patients, catheter intervention procedure triggered CCE (catheter-related CCE). In the remaining 3 patients, CCE spontaneously occurred after introduction of unfractionated heparin therapy during treatment of heart failure with myocardial infarction (spontaneous CCE). Age in spontaneous CCE (88+/-7 y) was significantly higher (p!0.05) than that in catheter-related CCE (71+/-2 y). Blue toe was a first sign to diagnose CCE in ll patients. Conclusions: Our data indicated that 1) frequency of spontaneous CCE is not low compared to that of catheter-related CCE, 2) spontaneous CCE may be an important comorbidity of heart failure with myocardial infarction, 3) advanced age may be a risk of spontaneous CCE, and 4) blue toe is the most important sign for management of CCE.

P-024 Acute Effect of Additional Administration of High-dose Nicorandil for Patients with Acute Myocardial Infarction after Successful PCI SHINJI HISATAKE, TAKAYUKI KABUKI, SHUNSUKE KIUCHI, YUICHIRO FUJII, TOSHIO KINOSHITA, TAKATOSHI IKEDA, JUNICHI YAMAZAKI Department of Cardiovascular Medicine, Toho University Omori Medical Center, Tokyo, Japan Background: Because reperfusion injury or LV remodeling leads to poor prognosis, reducing the infarction size is still an important issue. We examined the effectiveness of administration of carperitide and/or nicorandil immediately after PCI. Methods: Among patients who were hospitalized due to first-time acute anterior wall infarction, the consecutive 41patients with TIMI III by PCI became the subject of this study. The patients were divided into the four groups; control group, low-dose (0.0125-0.025g) carperitide administration group (C), high-dose (0.05-0.2mg/kg/h) nicorandil administration group (N), low-dose carperitide and high-dose nicorandil combined administration group (CN). Immediately after PCI, each drug was administrated for 72hours. CPK was measured before and after PCI, every 4hours up to 24hours after PCI, then 48hours and 72hours after PCI and sCPK was calculated. BNP was measured before and after PCI, 24hours, 48hours, and 72hours after PCI. Results: The control group and the other three groups showed no significant difference in sCPK and BNP, however, sCPK in N was significantly lower (p50.022), and BNP measured 24, 48 and 72hours after PCI in NS each showed a significantly smaller amount (p50.43, p50.44, p50.31) compared to C. Conclusion: Low-dose administration of carperitide was not effective enough to reduce infarction size, but high-dose