Paclitaxel-induced endothelial dysfunction in living rats is prevented by nicorandil via reduction of oxidative stress.

Paclitaxel-eluting stents dramatically reduce rates of in-stent restenosis; however, paclitaxel is known to lead to endothelial dysfunction. Protective effects of nicorandil on paclitaxel-induced endothelial dysfunction by examining flow-mediated dilation (FMD) were investigated in anesthetized rats. After 7-day osmotic infusion of paclitaxel (5 mg/kg per day), FMD was measured by high-resolution ultrasound in the femoral artery of living rats. Paclitaxel significantly reduced FMD (21.6% ± 3.2% to 7.1% ± 1.7%); this reduction was prevented by co-treatment with nicorandil (15 mg/kg per day), while paclitaxel did not affect nitroglycerin-induced vasodilation. Diazoxide and tempol, but not isosorbide dinitrate, had an effect similar to nicorandil in preventing paclitaxel-induced decrease in FMD. Nicorandil significantly prevented paclitaxel-induced reduction in acetylcholine-induced vasodilation. On the underling mechanisms, paclitaxel increased reactive oxygen species (ROS) production (dihydrorhodamine 123, DCF fluorescence intensity) and NADPH oxidase (p47(phox), gp91(phox) mRNA) in arteries and human coronary artery endothelial cells (HCAECs), while paclitaxel reduced nitric oxide (NO) release (DAF-2 fluorescence intensity), but not endothelial NO synthase (eNOS) phosphorylation in HCAECs. Nicorandil prevented the increased ROS production in arteries and HCAECs, which was 5-hydroxydecanoate (5-HD)-sensitive but 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ)-resistant, without significant effect on the reduced NO release. In conclusion, nicorandil prevents paclitaxel-induced endothelial dysfunction, which may be brought by improved NO bioavailability due to the reduction of oxidative stress via K(ATP) channel activation.

Cilostazol protects against microvascular brain injury in a rat model of type 2 diabetes.

Cilostazol, a pluripotent phosphodiesterase III-specific inhibitor with anti-platelet and vasculogenic effects, is useful for preventing recurrent brain vascular events, particularly in stroke patients with diabetes mellitus (DM). However, it is unclear whether cilostazol affects autoregulatory responses in small cerebral arteries. Thus, we investigated the effect of cilostazol on diabetic brain vasculopathy in a model of type II DM using male OLETF rats. OLETF rats were treated with either cilostazol (CG) or vehicle (VG) and subjected to microangiography with monochromatic synchrotron radiation to investigate middle cerebral artery (MCA) vasoreactivity following an injection of acetylcholine (Ach). Ach administration led to MCA diameter contraction in the VG, but MCA dilation in the CG. We also evaluated morphological changes in the small intracranial vessels and found that in the CG, the endothelial cell structure in the small artery was not destroyed. Moreover, protein levels of phosphorylated endothelial nitric oxide synthase (eNOS) and vascular endothelial growth factor (VEGF) were higher in each evaluated brain region in CG rats vs. VG rats. Our results suggest that cilostazol could potentially improve autoregulatory responses in the small cerebral arteries by increasing eNOS phosphorylation and VEGF expression in DM, and thus, may act as a neurovascular protectant.

Altered blood flow in cerebral perforating arteries of rat models of diabetes: A synchrotron radiation microangiographic study toward clinical evaluation of white matter hyperintensities.

AIM: As altered blood flow in the cerebral perforating arteries (PA) might be related to development of cerebral white matter hyperintensities, we examined whether the hemodynamic relationship of the PA and middle cerebral artery (MCA) is altered in rat models of diabetes, compared with normal rats and a rat model of sinoatrial denervation (blood pressure fluctuation model). METHODS: We used microangiography with monochromatic synchrotron radiation to measure the diameters of the PA and MCA at 4.5 µm resolution in five groups of rats: (i) Long-Evans Tokushima Otsuka (LETO); (ii) Otsuka Long-Evans Tokushima Fatty (a model of type 2 diabetes with obesity); (iii) LETO with sinoaortic denervation (LETO + SAD); (iv) F344; and (v) F344 + streptozotocin (a model of type 1 diabetes). RESULTS: Compared with LETO, Otsuka Long-Evans Tokushima Fatty rats showed a significant reduction in the diameter of both PA and MCA, though the PA/MCA diameter ratio was unchanged. In contrast, compared with LETO, LETO + SAD rats showed an increased MCA diameter, and the PA/MCA diameter ratio was decreased. Compared with F344 rats, the MCA diameter was increased in F344 + streptozotocin rats, and the PA/MCA diameter ratio was decreased. Scatter diagrams showed that the diameters of the PA and MCA were essentially independent of each other in the two types of diabetic models. CONCLUSION: PA were consistently visualized at high resolution by means of microangiography using synchrotron radiation. The present results show that rat diabetic models exhibit changes in PA diameter and PA/MCA diameter ratio, which might be related to the development of diabetes-associated cerebral white matter hyperintensities.

Minimal inspiratory flow from dry powder inhalers according to a biphasic model of pressure vs. flow relationship.

Inhalation therapy using the dry powder inhaler (DPI) is now the first choice for obstructive pulmonary diseases. We previously measured relationships between inspiratory pressure (PI) and flow rate of almost all of the DPIs available in Japan, and described an importance of inspiratory efforts. In the present study, we further analyzed the data obtained in the previous study. Although there were linear relationships between PI and flow2, the slope became steeper when PI was less than a certain value (critical PI, existed between 15-20 cmH2O). When PI was less than critical PI, linear rather than parabolic regression between PI and flow yielded better fits (r > 0.90, p < 0.001). Inspiratory flows at the critical PI were 53.9 (Diskus), 65.8 (Diskhaler), 45.9 (Turbuhaler for Pulmincort), 48.6 (Turbuhaler for Symbicort) and 38.0 l/min (Twisthaler). These findings suggested that flow through the DPI becomes laminar rather than turbulent flow in the range below critical PIs. We suggest that patients should inhale from the DPIs with inspiratory pressure higher than critical PI.

Possible mechanism of preventive effects of coffee intake on the formation of arterial occlusive thrombosis.

BACKGROUND: Prevalence and incidence of arterial occlusive thrombosis are influenced by life-style. Coffee consumption was shown with a lower incidence of myocardial infarction by Framingham Study. Yet, the mechanism is to be elucidated. METHODS: We examined the effects of coffee intake on the progression of occlusive thrombus formation in mouse cremasteric arteries. After 7 days of free intake of pure water, coffee containing water (5 mg/ml), or caffeine containing water (0.1 mg/ml), endothelial cell function was locally damaged by FeCl3. Circulating platelet and leukocytes were rendered fluorescently by rhodamine 6G. Process of occlusive thrombus growth was continuously visualized by 3-D imaging system equipped with ultra-fast confocal microscopy, and time to vascular occlusion was measured in each mouse. RESULTS: Platelet accumulation started immediately after FeCl3 exposure in all tested groups. However, arterial occlusion time in taking coffee containing water was significantly longer than those taking pure water. (46.0 ± 17.4 min (n = 5) vs. 12.3 ± 2.6 min (n = 31), p < 0.05) Arterial occlusion time in mice taking caffeine (13.8 ± 5.9 min (n = 4)) was not different from those taking pure water. CONCLUSION: Coffee, but not caffeine intake, may have preventive effect on arterial occlusive thrombus formation initiated by functional injury of arterial endothelium.

Imaging of structural changes in endothelial cells and thrombus formation at the site of FeCl(3)-induced injuries in mice cremasteric arteries.

AIM: We investigated thrombus formation at the site of functional injury to endothelial cells by FeCl(3). METHODS: After preparation of cremasteric arteries of mice, controlled endothelial injury was induced by application of FeCl(3). Endothelial cells were rendered fluorescent by addition of FITC (fluorescein isothiocyanate)-labeled isolectin B4. Circulating platelets and leukocytes were made fluorescent by rhodamine 6G. Three-dimensional (3D) growth of thrombi was visualized in real time. Effects of aspirin and clopidogrel pre-treatments on the growth of thrombi were investigated in vivo as well as in an ex vivo flow chamber system. RESULTS: Endothelial cells were tightly bound to each other to protect local thrombus formation. Platelets started to adhere to endothelial cells when FeCl(3) was applied. Three-dimensional growth of thrombi, which takes 10.6+/-7.5 minutes for complete occlusion in control, can be visualized with our imaging system. Aspirin pre-treatment at the dose tested did not influence either endothelial injury or platelet thrombus growth, while clopidogrel pretreatment significantly inhibited 3D growth and prolonged occlusion time up to 64.6+/-25.3 minutes (100 mg/kg). A similar inhibiting effect of clopidogrel was reproduced in ex vivo flow chamber experiments. CONCLUSIONS: We have developed an in vivo system to detect thrombus formation after functional damage to the endothelium.