Vascular Apoptosis Research Articles

Unseeded Inertial Cavitation for Enhancing the Delivery of Chemotherapies: A Safety Study

Acoustic cavitation can improve local drug delivery in tumors. Without injected external nucleation agents, initiating inertial cavitation requires high negative pressures, which can lead to biological damage. In the present study, unseeded inertial cavitation was obtained in vivo using confocal beams, and the effect of these exposure conditions was assessed on drug structure and activity, shallow tissues and growth of breast tumors. No change was observed in the structure and cytotoxicity of doxorubicin. Experiments were conducted on healthy rats, exposing the thigh and abdomen. Histologic analyses at 72 h and 2 weeks post-treatment demonstrated a modest impact on tissues. Syngeneic 4 T1 breast tumors in mice were sonicated. Immunohistochemical analyses showed that ultrasound did not impact vascular density, proliferation and apoptosis of cancer cells. In addition, ultrasound did not negatively modify cancer cell spreading to the lungs and bone marrow. This provides evidence that these particular parameters can be used safely in vivo.

Food supplementation with rice bran enzymatic extract prevents vascular apoptosis and atherogenesis in ApoE-/- mice.

Atherosclerosis is associated with reduced mononuclear cell (MNC) telomere length, and senescent cells have been detected in atherosclerotic plaques. Rice bran is a source of γ-oryzanol, phytosterols and tocols with potential lipid-lowering, antioxidant and anti-inflammatory activities. Here, we tested the hypothesis that rice bran enzymatic extract (RBEE) impacts on apoptosis, telomere length and atherogenesis in mice. Seven-week-old male ApoE-/- mice were fed high-fat diet (HFD) or isocaloric HFD supplemented with 5% (w/w) RBEE for 23weeks. Wild-type mice of the same age were kept under standard diet as controls. RBEE treatment reduced total cholesterol (19.24±1.63 vs 24.49±1.71mmol/L) and triglycerides (1.13±0.18 vs 1.75±0.22mmol/L) and augmented HDL-cholesterol (1.86±0.20 vs 1.07±0.20mmol/L). RBEE attenuated macrophage infiltration by 56.69±4.65% and plaque development (7737±836 vs 12,040±1001μm2) in the aortic sinus. In the aorta, RBEE treatment reduced expression of the apoptosis pathway components p16, p53 and bax/bcl-2 ratio. RBEE prevented apoptosis of aortic endothelial cells (2.81±0.71-1.14±0.35 apoptotic nuclei/ring for ApoE-/- HFD and ApoE-/- HFD 5% RBEE, respectively). In contrast, MNC of RBEE-fed mice exhibited enhanced apoptosis marker expression with increased p53 and bax/bcl-2 protein levels. Compared to WT, ApoE-/- mice on HFD were characterized by significant telomere shortening in aorta (11±2%) and MNC (73±7%), which was reduced by supplementation with RBEE (aorta: 40±7%; MNC: 105±10%). Expression of telomere repeat-binding factor 2 was increased in RBEE-fed mice. Long-term food supplementation with RBEE lowers cholesterol and prevents atherosclerotic plaque development in ApoE-/- mice. Differential regulation of vascular and MNC apoptosis and senescence were identified as potential mechanisms.

Abstract 005: Axl Controls Survival of the CD4+ T Lymphocytes in Salt-dependent Hypertension

Introduction: Axl, a receptor tyrosine kinase, is required for vascular and immune cell survival. We sought to investigate the effects of Axl on T lymphocyte survival during deoxycorticosterone acetate (DOCA)-salt hypertension in mice. Methods and Results: We found significant reduction in systolic blood pressure (BP) after 5-6 weeks of DOCA-salt in RAG1-/- mice after adoptive transfer of CD4+ T cells from Axl knockout (Axl-/- →RAG1-/-) compared to transferred CD4+ T cells from wild type (Axl+/+ →RAG1-/-) mice. Media area of the mesenteric artery was significantly lower in Axl-/- →RAG1-/- (4.2±0.7x10 3 m 2 ) vs. Axl+/+ →RAG1-/- (6.0±0.9x10 3 m 2 ) or Axl+/+ (6.8±0.6x10 3 m 2 ) mice. There was significant decrease in interferon gamma production by the T cells from Axl-/- (396±23 ng/mL) compared to Axl+/+ (512±42 ng/mL) after T h 1-priming. The number of carboxyfluorescein succinimidyl ester-positive cells in 6 th division was dramatically declined in Axl-/- (~0.3%) vs. Axl+/+ (~1.8%) in culture. Accordingly, we found lower number of lymphocytes in blood from Axl-/- (4.5±0.7x10 9 ) compared to Axl+/+ (7.8±0.7x10 9 ) mice. Blood leukocyte apoptosis was 2.5-fold higher in Axl-/- mice. We next investigated repopulation capacities of the hematopoietic cells from Axl-/- vs. Axl+/+ mice. There was significant decrease in Axl-/- CD3+ T cells (21±3 %) than Axl+/+ (49±3 %) in spleen after 8 weeks of competitive repopulation of bone marrow-derived cells. However, we found even greater reduction of Axl-/- T lymphocytes (15±1 %) vs. Axl+/+ T lymphocytes (52±6 %) in peripheral blood after 8 weeks of competitive repopulation. Finally, percentage of apoptotic cells was the greatest in the media (20±7 %) and adventitia (13±5 %) from Axl-/- →RAG1-/- mice compared to vascular apoptosis (6-14 % in media; and 6-9 % in adventitia) in other groups after 6 weeks of DOCA-salt. Conclusions: Our data suggest that Axl-dependent survival of the T lymphocytes is crucial for the late increase in BP in DOCA-salt hypertension.

Chemerin Regulates Crosstalk Between Adipocytes and Vascular Cells Through Nox.

Adipocytes produce adipokines, including chemerin, a chemoattractant that mediates effects through its ChemR23 receptor. Chemerin has been linked to endothelial dysfunction and vascular injury in pathological conditions, such as obesity, diabetes mellitus, and hypertension. Molecular mechanisms underlying this are elusive. Here we assessed whether chemerin through redox-sensitive signaling influences molecular processes associated with vascular growth, apoptosis, and inflammation. Human microvascular endothelial cells and vascular smooth muscle cells were stimulated with chemerin (50 ng/mL). Chemerin increased generation of reactive oxygen species and phosphorylation of mitogen-activated protein kinases, effects that were inhibited by ML171, GKT137831 (Nox inhibitors), and N-acetylcysteine (reactive oxygen species scavenger). Chemerin increased mRNA expression of proinflammatory mediators in vascular cells and increased monocyte-to-endothelial cell attachment. In human vascular smooth muscle cells, chemerin induced phosphorylation of mitogen-activated protein kinases and stimulated proliferation (increased proliferating cell nuclear antigen expression [proliferation marker] and BrdU incorporation [proliferation assay]). Chemerin decreased phosphatidylinositol 3-kinase/protein kinase B activation and increased TUNEL-positive human vascular smooth muscle cells. In human microvascular endothelial cells, chemerin reduced endothelial nitric oxide synthase activity and nitric oxide production. Adipocyte-conditioned medium from obese/diabetic mice (db/db), which have elevated chemerin levels, increased reactive oxygen species generation in vascular smooth muscle cells, whereas adipocyte-conditioned medium from control mice had no effect. Chemerin actions were blocked by CCX 832, a ChemR23 inhibitor. Our data demonstrate that chemerin, through Nox activation and redox-sensitive mitogen-activated protein kinases signaling, exerts proapoptotic, proinflammatory, and proliferative effects in human vascular cells. These findings elucidate some molecular mechanisms through chemerin, which is increased in obesity, whereby adipocytes may influence vascular function. We identify chemerin as a novel vasoactive adipokine, which may be important in obesity-related vascular injury.

Molecular magnetic resonance imaging of atherosclerotic vessel wall disease.

Molecular imaging aims to improve the identification and characterization of pathological processes in vivo by visualizing the underlying biological mechanisms. Molecular imaging techniques are increasingly used to assess vascular inflammation, remodeling, cell migration, angioneogenesis and apoptosis. In cardiovascular diseases, molecular magnetic resonance imaging (MRI) offers new insights into the in vivo biology of pathological vessel wall processes of the coronary and carotid arteries and the aorta. This includes detection of early vascular changes preceding plaque development, visualization of unstable plaques and assessment of response to therapy. The current review focuses on recent developments in the field of molecular MRI to characterise different stages of atherosclerotic vessel wall disease. A variety of molecular MR-probes have been developed to improve the non-invasive detection and characterization of atherosclerotic plaques. Specifically targeted molecular probes allow for the visualization of key biological steps in the cascade leading to the development of arterial vessel wall lesions. Early detection of processes which lead to the development of atherosclerosis and the identification of vulnerable atherosclerotic plaques may enable the early assessment of response to therapy, improve therapy planning, foster the prevention of cardiovascular events and may open the door for the development of patient-specific treatment strategies. Targeted MR-probes allow the characterization of atherosclerosis on a molecular level. Molecular MRI can identify in vivo markers for the differentiation of stable and unstable plaques. Visualization of early molecular changes has the potential to improve patient-individualized risk-assessment.

FOXO3A elicits vascular smooth muscle cell apoptosis through release of MMP-13 and subsequent degradation of the extracellular matrix

FOXO3A elicits vascular smooth muscle cell apoptosis through release of MMP-13 and subsequent degradation of the extracellular matrix

Embryonic Stem Cells and Released Factors Stimulate c-kit+/FLK-1+ Progenitor Cells and Promote Neovascularization in Doxorubicin-Induced Cardiomyopathy

Vascular apoptosis plays a pivotal role in the development and progression of a myriad of cardiac dysfunctions, but has yet to be investigated in doxorubicin-induced cardiomyopathy (DIC). Additionally, the neovascularization potential and resulting functional consequences of embryonic stem (ES) cells and factors released from these cells in the chronic DIC myocardium remain largely unknown. To this end, we transplanted conditioned media (CM) and ES cells in the DIC-injured heart and evaluated their potential to inhibit vascular cell death, activate endogenous c-kit(+) and FLK-1(+) cells, enhance neovascularization, and augment left ventricular dysfunction. Data presented suggest transplanted CM and ES cells significantly blunt vascular cell apoptosis consequent to DIC. Quantitative immunohistochemistry data demonstrate significantly increased c-kit(+) and FLK-1(+) cells, as well as enhanced differentiated CD31(+) cells in the CM and ES cell groups relative to DIC controls. Heart function, including fractional shortening and ejection fraction, assessed by transthoracic echocardiography, was significantly improved following CM and ES cell transplantation. In conclusion, our data suggest that transplantation of CM and ES cells inhibit vascular apoptosis, activate endogenous c-kit(+) and FLK-1(+) cells and differentiate them into endothelial cells, enhance neovascularization, and improve cardiac function in the DIC-injured myocardium.

Magnetic resonance imaging of spinal trauma at acute and early stage

The topic actuality is conditioned upon the necessity to optimize the diagnostic detection in patients with spinal injury, imaging of spinal medulla damage, secondary alterations with the help of MRI examination. Standard magnetic resonance imaging examination of the patients was performed to diagnose the prevalence rate of ischemia and edema. According to previously published studies, the main factors in the pathogenesis of spinal cord injury are attributed to primary and secondary mechanism. Primary spinal cord injury occurs at the time of the initial trauma or damage, forming a zone of spinal cord contusion. Disorders can be found in the local vasculature, causing edema and hemorrhage. Paralysis of motor, sensory and autonomic neuronal functions can occur in the damaged area. Vascular dysfunction, ischemia, glutamatergic excitotoxicity, inflammation and apoptosis also develop in the perifocal zone.Methods. There was carried out standard MRI examination of the affected region of vertebral column in 24 patients, 11 patients of which has trauma in cervical area and 13 – in thoracic part. The examination was carried out within the term from 1 to 18 days, in the modes Т1 and Т2 WI in IS SE. To standardize the study there was defined extension of edema or signal change in the medullary substance, level of spinal medulla compression.Results. The study is carried out on the following day after trauma and it is defined the site of injury, hemorrhachis and medullary substance edema site in the area of initial injury within the measures of 1-2 segments of spinal medulla. MRI of 2-13 days shows increased distribution of hemorrhachis and medullary substance edema within 4-5 segments of spinal medulla, evidencing globalization of secondary alteration of spinal medulla.Conclusions. Magnetic resonance imaging is the leading method in the diagnosis of swelling of the spinal cord injury and hemorrhage. Secondary changes in the spinal cord are revealed during the study. The method allows visualizing the extent and severity of damage to the spinal cord, hence the importance of the method in the initial phase of treatment, patients with spinal cord injury

Evidence that Vascular Endothelial Derived Endothelin‐1 Promotes Development of Tunicamycin‐Induced Endoplasmic Reticulum Stress in Renal Vessels

Levels of the vasoconstrictor peptide endothelin-1 (ET-1) are elevated in cardiovascular disease. Endoplasmic reticulum (ER) stress is one of the known mechanisms for cellular apoptosis. Our group has recently implicated the ET-1 system in the activation of renal ER stress pathways, although it is unclear which cells are involved. To clarify the role of ET-1 in the development of renal vascular ER stress, we hypothesized that lack of ET-1 in endothelial cells attenuates tunicamycin (TM)-induced ER stress in renal vessels. Female mice lacking ET-1 in vascular endothelial cells (VEET KO) and the corresponding flox controls (n=5-9/group) received a single i.p. injection of either TM (1.5 mg/kg) or saline. 24 hours post-injection, renal vessels were isolated, ER stress genes determined by qRT-PCR, and renal apoptosis detected by TUNEL staining. TM significantly increased the expression of ER stress markers in renal vessels from flox mice (saline vs. TM, fold change/flox+saline, p<0.05; GRP94: 1.0±0.3 vs. 16.5±6.6, ATF-6: 1.0±0.5 vs. 6.8±3.5, and CHOP: 1.4±0.4 vs. 12.5±1.9). TM treatment led to increased outer medullary, non-vascular TUNEL staining in flox mice as well. TM failed to increase renal vascular ER stress markers or renal apoptosis in VEET KO mice. These results indicate that ET-1 is critical for the development of renal vascular ER stress and apoptosis in response to TM, and highlight the possibility of targeting the ET-1 system as a therapeutic approach against ER stress-induced renal vascular damage. Funded by NIH T32 DK007545 to CDM and P01 HL95499 to DMP and JSP.

Snake venom derived molecules in tumor angiogenesis and its application in cancer therapy; an overview.

Snake venom is a complex mixture of biologically and pharmacologically active components, comprising hydrolytic enzymes, non-enzymatic proteins/peptides, and small amounts of organic and inorganic molecules. The venom components are known to vary with geographic location, season, species and age of the snakes. The role of the venom in the snake is not primarily for self-defense, but in prey immobilization and its subsequent digestion. Hence, several digestive enzymes in venoms, in addition to their hydrolytic activity have evolved to interfere in diverse physiological processes that help in the immobilization of prey/victim. As snake components are capable of modulating the physiological response of envenomated prey/victim, they show promise as potential pharmacological tools, as drug leads and in diagnostic applications. This, in a practical sense to be a reality has to be linked to the advances in toxinology that provide investigators with an understanding of the pharmacodynamics of toxins together with improved understanding of the etiology of many human diseases and identification of potential sites for therapeutic intervention. This review aims at providing an overview on snake venom toxins and their derivatives that have potential anti-angiogenic effects for cancer treatment. Some of the anti-angiogenic components of snake venom like Snake venom metalloproteinases (SVMPs), Disintegrins, Phospholipases A2 (PLA2), CType Lectins (CLP), Vascular Apoptosis inducing Proteins (VAP) and L-Amino Acid Oxidases (LAAO) are discussed. This review aims at giving an overall view of these molecules and their mechanism of action as an effective antiangiogenic agent towards the treatment of cancer.

Vascular endothelial injury and apoptosis in rats with severe acute pancreatitis.

We explored mechanisms of vascular endothelial injury that lead to systemic multiple organ failure by detecting the soluble endothelial protein C receptor (sEPCR), von Willebrand factor (vWF), serum nitric oxide (NO), and tumor necrosis factor alpha (TNF-α) and Bcl-2 mRNA and Bax mRNA expression in a severe acute pancreatitis (SAP) rat model. Compared to controls, the levels of TNF-α, vWF, and sEPCR were significantly increased in the experimental group at 12 hours and 24 hours and the NO level was significantly decreased. After 12 hours, the aortic endothelial apoptosis index and Bax mRNA expression in aortic endothelial cells had increased in the experimental group, but Bcl-2 mRNA levels had decreased. All these changes appeared at both 12 h and 24 hours. The results indicated that vascular endothelial injury and apoptosis markers were elevated in SAP. Endothelial injury and increased apoptosis in the experimental group were related to the increased expression of TNF-α.

Epicatechin breaks preformed glycated serum albumin and reverses the retinal accumulation of advanced glycation end products

The accumulation of advanced glycation end products (AGEs) is associated with many of the complications of diabetes mellitus, including diabetic retinopathy. AGE-breakers, such as N-phenacylthiazolium and alagebrium, have been proposed as therapeutic agents for reversing the increase in protein crosslinking in diabetes. (−)-Epicatechin is a major dietary flavonoid with a wide range of health-promoting biological activities. The aim of this study was to determine the potential effect of (−)-epicatechin in reducing the burden of AGEs in vitro and in vivo and to evaluate whether the reduced AGE burden could translate into improvement in retinal vascular function in exogenously AGE-injected rats. Glycated human serum albumin was purified from patients with diabetes. The breakdown of the already formed AGEs was studied by treating glycated human serum albumin with (−)-epicatechin. To study the effect of (−)-epicatechin on retinal vascular function, exogenously AGE-injected rats were treated with (−)-epicatechin (50 and 100mg/kg i.p.) for two weeks. Apoptosis of retinal vascular cells was quantified using TUNEL staining. The AGE load in the retinas was determined via immunohistochemical staining and western blot analysis. (−)-Epicatechin was able to break preformed glycated human serum albumin in vitro as well as reduce AGE accumulation in retinas in vivo in a dose dependent manner. In exogenously AGE-injected rats, treatment with (−)-epicatechin was evidenced by an improved retinal vascular apoptosis. AGE burden in retinas was also reduced upon treatment. This study suggests that (−)-epicatechin could represent a valuable drug for the treatment of diabetic retinopathy by reducing the AGE burden.

Novel Peptide for Attenuation of Hyperoxia-induced Disruption of Lung Endothelial Barrier and Pulmonary Edema via Modulating Peroxynitrite Formation

Pulmonary damages of oxygen toxicity include vascular leakage and pulmonary edema. We have previously reported that hyperoxia increases the formation of NO and peroxynitrite in lung endothelial cells via increased interaction of endothelial nitric oxide (eNOS) with β-actin. A peptide (P326TAT) with amino acid sequence corresponding to the actin binding region of eNOS residues 326-333 has been shown to reduce the hyperoxia-induced formation of NO and peroxynitrite in lung endothelial cells. In the present study, we found that exposure of pulmonary artery endothelial cells to hyperoxia (95% oxygen and 5% CO2) for 48 h resulted in disruption of monolayer barrier integrity in two phases, and apoptosis occurred in the second phase. NOS inhibitor N(G)-nitro-L-arginine methyl ester attenuated the endothelial barrier disruption in both phases. Peroxynitrite scavenger uric acid did not affect the first phase but ameliorated the second phase of endothelial barrier disruption and apoptosis. P326TAT inhibited hyperoxia-induced disruption of monolayer barrier integrity in two phases and apoptosis in the second phase. More importantly, injection of P326TAT attenuated vascular leakage, pulmonary edema, and endothelial apoptosis in the lungs of mice exposed to hyperoxia. P326TAT also significantly reduced the increase in eNOS-β-actin association and protein tyrosine nitration. Together, these results indicate that peptide P326TAT ameliorates barrier dysfunction of hyperoxic lung endothelial monolayer and attenuates eNOS-β-actin association, peroxynitrite formation, endothelial apoptosis, and pulmonary edema in lungs of hyperoxic mice. P326TAT can be a novel therapeutic agent to treat or prevent acute lung injury in oxygen toxicity.

Hydrogen sulfide: metabolism, biological and medical role

Hydrogen sulfide (H2S) is a signaling molecule that is actively synthesized in the tissues and is involved in the regulation of vascular tone, neuromodulation, cytoprotection, inflammation and apoptosis. In recent years, new data on animal and human H2S metabolism and function under the effect of various endogenous and exogenous factors, including drugs were collected. This review is provided to introduce generalized information about the main and alternative H2S metabolism and regulation, peculiarities of transport, signaling, biological role and participation in pathogenesis. Submitted data describe H2S content and activity of H2S-synthesizing enzymes in different organs, H2S effect on blood coagulation and platelet aggregation based on our research results. The working classification of H2S metabolism modulators, which are used in biology and medicine, is proposed: 1) agents that increase H2S content in tissues (inorganic and organic H2S donors; H2S-synthesizing enzymes substrates and their derivatives, H2S-releasing drugs; agents that contain H2S-synthesizing enzymes cofactors and activators, agents that inhibit H2S utilization); 2) agents that reduce H2S content in tissues (specific and nonspecific inhibitors of H2S-synthesizing enzymes), 3) agents with uncertain impact on H2S metabolism (some medicines). It was demonstrated that vitamin-microelement and microelement complexes with H2S-synthesizing enzymes cofactors and activators represent a promising approach for H2S content correction in tissues.

Aortic and Carotid Arterial Stiffness and Epigenetic Regulator Gene Expression Changes Precede Blood Pressure Rise in Stroke-Prone Dahl Salt-Sensitive Hypertensive Rats

Multiple clinical studies show that arterial stiffness, measured as pulse wave velocity (PWV), precedes hypertension and is an independent predictor of hypertension end organ diseases including stroke, cardiovascular disease and chronic kidney disease. Risk factor studies for arterial stiffness implicate age, hypertension and sodium. However, causal mechanisms linking risk factor to arterial stiffness remain to be elucidated. Here, we studied the causal relationship of arterial stiffness and hypertension in the Na-induced, stroke-prone Dahl salt-sensitive (S) hypertensive rat model, and analyzed putative molecular mechanisms. Stroke-prone and non-stroke-prone male and female rats were studied at 3- and 6-weeks of age for arterial stiffness (PWV, strain), blood pressure, vessel wall histology, and gene expression changes. Studies showed that increased left carotid and aortic arterial stiffness preceded hypertension, pulse pressure widening, and structural wall changes at the 6-week time-point. Instead, differential gene induction was detected implicating molecular-functional changes in extracellular matrix (ECM) structural constituents, modifiers, cell adhesion, and matricellular proteins, as well as in endothelial function, apoptosis balance, and epigenetic regulators. Immunostaining testing histone modifiers Ep300, HDAC3, and PRMT5 levels confirmed carotid artery-upregulation in all three layers: endothelial, smooth muscle and adventitial cells. Our study recapitulates observations in humans that given salt-sensitivity, increased Na-intake induced arterial stiffness before hypertension, increased pulse pressure, and structural vessel wall changes. Differential gene expression changes associated with arterial stiffness suggest a molecular mechanism linking sodium to full-vessel wall response affecting gene-networks involved in vascular ECM structure-function, apoptosis balance, and epigenetic regulation.

Chronic cardiovascular disease-associated gene network analysis in human umbilical vein endothelial cells exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin.

The association of dioxin exposure with increased morbidity or mortality of chronic cardiovascular diseases (CVDs) has been established by many epidemiological studies. However, the precise global gene expression alterations caused by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in the cardiovascular system need to be further elucidated. In this study, we profiled the gene expression of human umbilical vein endothelial cells (HUVECs) exposed to different concentrations of TCDD by high-throughput sequencing. Expression of 1,838 genes was changed significantly after TCDD stimulation. The FunDO analysis suggested that some CVDs were highly associated with TCDD treatment, including atherosclerosis, thromboangiitis obliterans, pulmonary arterial hypertension (PAH), and hypertension. KEGG pathway analysis showed that many genes in the signaling pathways of vascular smooth muscle contraction and apoptosis were altered distinctly. In addition, we revealed evidence regarding the gene network changes of chronic CVDs including atherosclerosis, thrombosis, myocardial infarction (MI), hypertension, and PAH in TCDD-exposed HUVECs. We found that gene expression of β1-adrenoceptors (ADRB1), β2-adrenoceptors (ADRB2), endothelin-converting enzyme 1 (ECE1), and endothelin-1 gene (EDN1) that are involved in the blood pressure regulation pathway decreased apparently under TCDD treatment. Moreover, the transcripts of interleukin 1 beta (IL-1β) and tumor necrosis factor α (TNFα), which are related to atherosclerosis, were up-regulated by TCDD stimulation. In addition, the transcripts of Homo sapiens collagen, type IV, alpha 1 (COL4A1), and isoforms that trigger the MI pathway were up-regulated after TCDD exposure. Finally, we found enhanced platelet-derived growth factor (PDGF) and signal transducer and activator of transcription 5 (Stat5) expression with TCDD treatment in endothelial cells, which are involved in PAH induced by vascular injury.

Age-dependent modifications in vascular adhesion molecules and apoptosis after 48-h reperfusion in a rat global cerebral ischemia model.

Stroke is one of the leading causes of death and permanent disability in the elderly. However, most of the experimental studies on stroke are based on young animals, and we hypothesised that age can substantially affect the stroke response. The two-vessel occlusion model of global ischemia by occluding the common carotid arteries for 15min at 40mmHg of blood pressure was carried out in 3- and 18-month-old male Sprague-Dawley rats. The adhesion molecules E- and P-selectin, cell adhesion molecules (CAMs), both intercellular (ICAM-1) and vascular (VCAM-1), as well as glial fibrillary acidic protein (GFAP), and cleaved caspase-3 were measured at 48h after ischemia in the cerebral cortex and hippocampus using Western blot, qPCR and immunofluorescence techniques. Diametric expression of GFAP and a different morphological pattern of caspase-3 labelling, although no changes in the cell number, were observed in the neurons of young and old animals. Expression of E-selectin and CAMs was also modified in an age- and ischemia/reperfusion-dependent manner. The hippocampus and cerebral cortex had similar response patterns for most of the markers studied. Our data suggest that old and young animals present different time-courses of neuroinflammation and apoptosis after ischemic damage. On the other hand, these results suggest that neuroinflammation is dependent on age rather than on the different vulnerability described for the hippocampus and cerebral cortex. These differences should be taken into account in searching for therapeutic targets.

P13.14 * PROPHYLACTIC USE OF SHIUNKO IN THE PREVENTION OF RADIATION-INDUCED SCALP DERMATITIS: A PRELIMINARY REPORT

INTRODUCTION: Radiation-induced scalp dermatitis, a complication of radiotherapy for brain tumors, may impair the quality-of-life (QOL) of patients. Steroid ointments have been considered as the only effective treatment, but the author had reported that shiunko,a sort of oriental ointments, could relief well the symptoms of radiation-induced scalp dermatitis [Kampo Med 62(2):142-146, 2011]. In this preliminary study, In this preliminary study, the author hypothesized that the prophylactic use of shiunko could prevent or alleviate radiation-induced scalp dermatitis. MATERIALS AND METHOD: Ten patients (male 4 and female 6, ranges between 21 to 79 years-old, mean 60.3 years-old,) with malignant brain tumors were involved in this study. Radiation dose ranges between 45 to 60 Gy, mean 54.7 Gy). Shiunko was applied to the whole radiated field consecutively during the entire period of radiotherapy. The efficacies were evaluated according to Common Terminology Criteria for Adverse Events (CTCAE) Version 4.0. RESULTS: Favorable therapeutic effects were observed in all patients. The scalp conditions were maintained well until the end of radiotherapy. The radiation dermatitis of all patients was estimated as Grade 1. But loss of hair was not avoided. DISCUSSION: Shiunko, which is covered by the Japanese health-insurance system for treating burns, is composed of only five natural remedies: lithospermi radix, angelicae radix, sesame oil, beeswax, and lard. Despite its simplicity, it has several unique activities, such as anti-bacterial activity, detoxification, anti-histamine activity, and moist wound healing. Furthermore, recent studies demonstrated vascular endothelial growth factor (VEGF) production and apoptosis induction, which promote wound healing. These preliminary results showed that shiunko is expected to be an effective ointment in preventing radiation-induced scalp dermatitis.

Cyclooxygenase-2-dependent oxidative stress mediates palmitate-induced impairment of endothelium-dependent relaxations in mouse arteries

Palmitic acid, one of the saturated free fatty acids, impairs cardiovascular function as manifested by inducing vascular inflammation, apoptosis and over-production of reactive oxygen species (ROS) although the origin for ROS remains unclear. The present study investigated the cellular mechanisms underlying palmitate-induced impairment of endothelial function. Ex vivo treatment in tissue culture with palmitate concentration-dependently attenuated acetylcholine-induced endothelium-dependent relaxations, up-regulated the expression of cyclooxygenase-2 (COX-2) and elevated superoxide formation in mouse aortic endothelial cells (MAECs) measured by dihydroethidium (DHE) staining and electron paramagnetic resonance (EPR) spectroscopy. Superoxide scavengers, COX-2 inhibitor and thromboxane prostanoid (TP) receptor antagonist, but not COX-1 inhibitor reversed the harmful effects of palmitate. Furthermore, palmitate impaired acetylcholine-induced relaxations and raised superoxide in en face endothelium of aortas only from COX-1−/− mice but not from COX-2−/− mice. Palmitate increased the production and release of TXB2, a stable thromboxane A2 metabolite in mouse aortas, which was abolished by COX-2 inhibitor. Superoxide scavenger did not affect palmitate-induced up-regulated expression of COX-2 in MAECs. Both real time PCR and luciferase reporter gene assay confirmed COX-2 up-regulation in palmitate-treated MAECs and NF-κB was substantially involved in this up-regulation. The present study provides novel evidence that palmitate up-regulates COX-2 through NF-κB-dependent mechanism and resultant COX-2-associated oxidative stress impairs endothelium-dependent relaxations in mouse aortas.

Microrna expression signature in human thoracic and abdominal aortic aneurysms

Objectives: MicroRNAs (miRNAs) are short non-coding RNAs that are post-transcriptional regulators of gene expression and some have been reported to be differentially expressed in thoracic aortic aneurysm (TAA); Ascending thoracic aneurysm (AscTAA) and Descending thoracic aneurysm (DTAA), abdominal aortic aneurysm (AAA), and carotid plaque (CP). Aneurysm (TAA, AAA) is a dilatation of the aorta, characterized by inflammation, vascular smooth muscle apoptosis, and extracellular matrix degradation. Fifteen candidate miRNAs implicated in TAA, AAA and CP were evaluated for their expression patterns in TAA, AAA, and CP and compared with normal control arteries.