Lapi Dominga, Chiurazzi Martina, Di Maro Martina, Mastantuono Teresa, Battiloro Laura, Sabatino Lina, Ricci Serena, Di Carlo Angelina, Starita Noemy, Guida Bruna, Santillo Mariarosaria, Colantuoni Antonio
Department of Clinical Medicine and Surgery, School of Medicine, University of Naples Federico II Naples, Italy.
Department of Science and Technology, University of Sannio Benevento, Italy.
Front Cell Neurosci. 2016 Jun 30;10:153. doi: 10.3389/fncel.2016.00153. eCollection 2016.
The present study was aimed to evaluate the malvidin's protective effects on damage induced by 30 min bilateral common carotid artery occlusion (BCCAO) and 60 min reperfusion (RE) in rat pial microcirculation. Rat pial microcirculation was observed using fluorescence microscopy through a closed cranial window. Western blotting analysis was performed to investigate the endothelial nitric oxide synthase (eNOS), phosphorylated eNOS (p-eNOS) and matrix metalloproteinase 9 (MMP-9) expression. Moreover, MMP-9 activity was evaluated by zymography. Finally, neuronal damage and radical oxygen species (ROS) formation were assessed. In all animals, pial arterioles were classified in five orders of branching according to Strahler's method. In hypoperfused rats, 30 min BCCAO and 60 min RE caused a decrease in arteriolar diameter, an increase in microvascular leakage and leukocyte adhesion, accompanied by decreased capillary perfusion and red blood cell velocity (VRBC). Moreover, marked neuronal damage and evident ROS generation were detected. Conversely, malvidin administration induced arteriolar dilation in dose-related manner, reducing microvascular leakage as well as leukocyte adhesion. Capillary perfusion and VRBC were protected. Nitric oxide (NO) synthase inhibition significantly attenuated malvidin's effects on arteriolar diameter. Western blotting analysis revealed an increase in eNOS and p-eNOS expression, while zymography indicated a decrease in MMP-9 activity after malvidin's administration. Furthermore, malvidin was able to prevent neuronal damage and to decrease ROS generation. In conclusion, malvidin protects rat pial microcirculation against BCCAO/RE injury, preventing blood-brain impairment and neuronal loss. Malvidin's effects appear to be mediated by eNOS activation and scavenger activity.
本研究旨在评估矢车菊素对大鼠软脑膜微循环中30分钟双侧颈总动脉闭塞（BCCAO）及60分钟再灌注（RE）所致损伤的保护作用。通过封闭的颅骨视窗，利用荧光显微镜观察大鼠软脑膜微循环。进行蛋白质免疫印迹分析以研究内皮型一氧化氮合酶（eNOS）、磷酸化eNOS（p-eNOS）和基质金属蛋白酶9（MMP-9）的表达。此外，通过酶谱法评估MMP-9活性。最后，评估神经元损伤和活性氧（ROS）的形成。在所有动物中，根据斯特勒尔方法将软脑膜小动脉分为五级分支。在灌注不足的大鼠中，30分钟的BCCAO和60分钟的RE导致小动脉直径减小、微血管渗漏和白细胞黏附增加，同时伴有毛细血管灌注减少和红细胞速度（VRBC）降低。此外，检测到明显的神经元损伤和明显的ROS生成。相反，矢车菊素给药以剂量相关方式诱导小动脉扩张，减少微血管渗漏以及白细胞黏附。毛细血管灌注和VRBC得到保护。一氧化氮（NO）合酶抑制显著减弱了矢车菊素对小动脉直径的影响。蛋白质免疫印迹分析显示，矢车菊素给药后eNOS和p-eNOS表达增加，而酶谱法表明MMP-9活性降低。此外，矢车菊素能够预防神经元损伤并减少ROS生成。总之，矢车菊素可保护大鼠软脑膜微循环免受BCCAO/RE损伤，预防血脑屏障损害和神经元丢失。矢车菊素的作用似乎是通过eNOS激活和清除剂活性介导的。
