434: Identifying PKC Epsilon as a Target Molecule To Control Intimal Hyperplasia (original) (raw)
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Circulation, 2004
Background-Inhibiting delta protein kinase C (␦PKC) during reperfusion and activating epsilon PKC (⑀PKC) before ischemia each limits cardiac ischemic injury. Here, we examined whether limiting ischemia-reperfusion injury inhibits graft coronary artery disease (GCAD) and improves murine cardiac allografting. Methods and Results-Hearts of FVB mice (H-2 q) were transplanted into C57BL/6 mice (H-2 b). ⑀PKC activator (⑀RACK) was injected intraperitoneally (20 nmol) into donor mice 20 minutes before procurement. Hearts were then perfused with ⑀RACK (1.5 nmol) through the inferior vena cava (IVC) and subsequently submerged in ⑀RACK (0.5 mol/L) for 20 minutes at 4°C. Before reperfusion, the peritoneal cavity of recipients was irrigated with ␦PKC inhibitor (␦V1-1, 300 nmol); control animals were treated with normal saline. The total ischemic time to the organ was 50 minutes. Two hours after transplantation, production of inflammatory cytokines and adhesion molecules, cardiomyocyte apoptosis, and caspase-3 and caspase-9 (but not caspase-8) activities were significantly reduced in the PKC regulator-treated group. Fas ligand levels (but not Fas) were also significantly reduced in this group. Importantly, GCAD indices, production of inflammatory cytokines, and adhesion molecules were significantly decreased and cardiac allograft function was significantly better as measured up to 30 days after transplantation. ConclusionsAn ⑀PKC activator and a ␦PKC inhibitor together reduced GCAD. Clinically, these PKC isozyme regulators may be useful for organ preservation and prevention of ischemia-reperfusion injury and graft coronary artery disease in cardiac transplantation. (Circulation. 2004;110[suppl II]:II-194-II-199.
Circulation, 2004
Background-Inhibiting delta protein kinase C (␦PKC) during reperfusion and activating epsilon PKC (⑀PKC) before ischemia each limits cardiac ischemic injury. Here, we examined whether limiting ischemia-reperfusion injury inhibits graft coronary artery disease (GCAD) and improves murine cardiac allografting. Methods and Results-Hearts of FVB mice (H-2 q ) were transplanted into C57BL/6 mice (H-2 b ). ⑀PKC activator (⑀RACK) was injected intraperitoneally (20 nmol) into donor mice 20 minutes before procurement. Hearts were then perfused with ⑀RACK (1.5 nmol) through the inferior vena cava (IVC) and subsequently submerged in ⑀RACK (0.5 mol/L) for 20 minutes at 4°C. Before reperfusion, the peritoneal cavity of recipients was irrigated with ␦PKC inhibitor (␦V1-1, 300 nmol); control animals were treated with normal saline. The total ischemic time to the organ was 50 minutes. Two hours after transplantation, production of inflammatory cytokines and adhesion molecules, cardiomyocyte apoptosis, and caspase-3 and caspase-9 (but not caspase-8) activities were significantly reduced in the PKC regulator-treated group. Fas ligand levels (but not Fas) were also significantly reduced in this group. Importantly, GCAD indices, production of inflammatory cytokines, and adhesion molecules were significantly decreased and cardiac allograft function was significantly better as measured up to 30 days after transplantation. ConclusionsAn ⑀PKC activator and a ␦PKC inhibitor together reduced GCAD. Clinically, these PKC isozyme regulators may be useful for organ preservation and prevention of ischemia-reperfusion injury and graft coronary artery disease in cardiac transplantation. (Circulation. 2004;110[suppl II]:II-194-II-199.)
LF 08-0299 In the prophylaxis and treatment of chronic rejection in a rat aortic allograft model
Transplant International, 2000
Abstract Chronic rejection is the major cause of late kidney allograft failure. We evaluated the efficacy of LF 08-299 (LF), an analogue of 15deoxyspergualin, in a rat aortic allograft model of chronic rejection. BN aortic allografts were transplanted to Lew recipients. LF was administered at a dose of 6 mg/kg and 2.5 mg/kg on days 0-20 and 6 mg/kg on days 60-90. CyA was used at a dose of 5 mg/kg on days 0-20. Untreated isografts and allografts were used as controls. Histological changes and immunohistochemistry were monitored sequentially at 8,12,16 and 20 weeks. There were no differences in intimal proliferation between LF-treated allografts and untreated or CyAtreated controls. Only a tendency in adventitial infiltration reduction was seen in LF-treated animals. We found a significantly less pronounced reduction in media diameter in LF-treated animals. We concluded that LF OW299 is only able to reverse reduction in media thickness in aortic allografts, but not inti-ma1 proliferation in this model of chronic rejection.
Circulation, 2010
Background-Protein kinase C (PKC) is involved in vascular smooth muscle cell (VSMC) activation, but little is known about its function in vascular pathology. We aimed at assessing the role of PKC in the development of restenosis. Methods and Results-Rat models of aortic balloon injury with or without subsequent stenting were used. Rats were treated with the selective PKC activator receptor for activated protein kinase C (RACK), the selective PKC inhibitor V1-2, or saline. Both down-stream cascades of the platelet-derived growth factor receptor via extracellular signal-regulated kinase and Akt, respectively, were evaluated in vivo and in VSMC cultures. Intimal hyperplasia with luminal obliteration developed in saline-treated balloon-injured rat aortas (20.3Ϯ8.0%), and RACK significantly promoted neointima development (32.4Ϯ4.9%, Pϭ0.033), whereas V1-2 significantly inhibited luminal narrowing (9.2Ϯ4.3%, Pϭ0.039). PKC inhibition led to significantly reduced VSMC extracellular signal-regulated kinase phosphorylation in vivo, whereas Akt phosphorylation was not markedly affected. Neointimal proliferation in vivo and platelet-derived growth factor-induced VSMC proliferation/migration in vitro were significantly inhibited by V1-2.
Gastroenterology, 2000
Background & Aims: Although protein kinase C (PKC) alterations may play a role in the abnormal reactivity of cirrhotic rat aortas, its isoforms and cellular distribution are unknown. We therefore studied the protein expression and cellular distribution of PKC isoforms and their activation in cirrhotic rat aortas. Methods: Endothelium-denuded aortas from control and cirrhotic rats were examined. Immunoblots were performed with PKC isoform-specific antibodies. Aortic reactivity was determined for phorbol myristate acetate and phenylephrine after PKC down-regulation. Results: PKC-␣ expression was reduced in both the cytosolic and membrane fractions in cirrhotic aortas. Trace amounts of PKC- were detected in cirrhotic aortas. PKC-␦ was detected in the cytosolic fraction of control and cirrhotic aortas. PKC-was detected in the membrane fraction in control aortas and in the cytosolic fraction in cirrhotic aortas. Phorbol myristate acetate and phenylephrine triggered translocation of PKC-␣ and PKC-␦ isoforms from the cytosol to the membrane in control aortas; in cirrhotic aortas, only PKC-␣ was translocated. Aortic reactivities were reduced after PKC downregulation. PKC-␣ and -␦ activities were reduced in cirrhotic aortas. Conclusions: These results suggest that a change in PKC isoforms may be responsible in part for the abnormal reactivity and intracellular transduction through the PKC pathway in cirrhotic rat aortas.
Biomedicine & Pharmacotherapy, 2019
Aortic stenosis (AS) is considered to be an actively regulated progress that involves similar pathophysiological processes as atherosclerosis. I-κB kinase-ε (IKKε) is a proinflammatory molecule involved in atherosclerosis. The objective of the present study was to define the role of IKKε in pathological valvular remodeling. Aortic valves (AVs) from 52 patients undergoing AV replacement (AS) and 13 patients undergoing heart transplant (Control) were analyzed. ApoE −/− mice (AK, n = 20) and ApoE −/− IKKε-/− mice (DK, n = 20) were generated and infused with saline or Ang II for 4 weeks. We found an upregulation of IKKε in human stenotic aortic valves compared to that in control AVs. Our results demonstrated that AK mice receiving AngII exhibited more advanced valvular remodeling and markedly increased IKKε expression. Conversely, loss of IKKε reduced adverse aortic valve thickening in response to Ang II, as measured by histological analyses. Furthermore, according to immunofluorescence analysis, Ang II resulted in obvious increases in the expression of α-SMA, TGF-β and NF-κB pathway components in the AK group, especially in the thickened area, while these increases were blocked in the DK group. Moreover, IKKε was co-expressed with α-SMA in valvular interstitial cells in ApoE −/− mice after an AngII infusion. These data provide evidence that IKKε plays a key role in the development of valvular remodeling and that it may be a novel target for the treatment of AS.
Circulation Research, 2000
Although systemic administration of NO donors has been shown to attenuate the development of neointimal hyperplasia in the balloon injury model, this strategy has not been tested in a model of allograft vasculopathy. In this study, we investigated the effect of FK409, a spontaneous NO releaser, on the development of allograft vasculopathy, using a rat aortic transplant model. Thoracic aortas from ACI rats were transplanted heterotopically into the abdominal aorta of Wistar-Furth rats. Postoperatively, recipients received FK409 orally every 8 hours from the day of transplantation to the time of euthanization. Morphometric and immunohistochemical analyses were performed on the aortic grafts 8 weeks after transplantation. Control allografts showed severe neointimal hyperplasia, which consists mainly of ␣-actincontaining vascular smooth muscle cells. The FK409-treated allografts showed a dose-dependent reduction (statistically significant compared with the control) in the neointimal thickness as the dose increased from 1 to 10 mg/kg (thrice per day). However, there was no significant difference in the neointimal thickness between groups treated with 10 and with 20 mg/kg. FK409 treatment (10 mg/kg) caused a significant decrease in DNA synthesis (5-bromo-2-deoxyuridine [BrdU] uptake), an increase in DNA fragmentation (terminal deoxynucleotidyltransferasemediated uridine nick-end labeling [TUNEL]), and upregulation of Fas expression, in the neointimal vascular smooth muscle cells. These data suggest that FK409 attenuates the allograft vasculopathy in a rat aortic transplant model. (Circ Res. 2000;87:66-72.