Vascular permeability of skeletal muscle microvessels in rat arterial ligation model: in vivo analysis using two-photon laser scanning microscopy
Vascular permeability of skeletal muscle microvessels in rat arterial ligation model: in vivo analysis using two-photon laser scanning microscopy
复制标题
大鼠动脉结扎模型中骨骼肌微血管的血管通透性：使用双光子激光扫描显微镜进行体内分析
DOI：
10.1152/ajpregu.00135.2020
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Kano Yutaka
中科院分区：
文献类型：
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作者：
Shimotsu Rie;Hotta Kzuki;Ikegami Ryo;Asamura Tomoyo;Tabuchi Ayaka;Masamoto Kazuto;Yagishita Kazuyoshi;Poole David C.;Kano Yutaka
下肢外周动脉疾病（PAD）由于动脉闭塞而影响氧气供应。缺血性骨骼肌伴有毛细血管结构变形。因此，使用新的显微镜技术，我们测试的假设，即内皮细胞肿胀时间和定量对应于增强微血管通透性。用髂动脉结扎法（AL）造成雄性Wistar大鼠后肢缺血.通过双光子激光扫描显微镜（TPLSM），使用静脉输注的罗丹明B异硫氰酸盐葡聚糖荧光染料对胫骨前肌（TA）微循环进行成像，并在AL后3天和7天定量染料外渗，以评估微血管通透性。透射电镜观察TA微血管内皮细胞超微结构。与对照组（0.40 ± 0.15 μ m3 × 106）相比，TPLSM测定的荧光染料在3（3.0 ± 0.40 μ m3 × 106）d和7（2.5 ± 0.8 μ m3 × 106）d的体积渗漏量增加（均P <0.05）。毛细血管壁厚度在第3天和第7天分别为（0.21 ± 0.06）μ m和（0.21 ± 0.08）μ m，与对照组（0.11 ± 0.03）μ m相比，差异均有统计学意义（P <0.05）。在PAD的AL模型中，毛细血管内皮细胞肿胀在时间和数量上与血管通透性升高相关，但这些变化发生在血管内皮生长因子（VEGF）及其受体（VEGFR2，AL-7天降低）或基质金属蛋白酶蛋白水平升高的情况下。内皮细胞肿胀和血管通透性增加的时间一致性支持一个共同的上游介质。TPLSM结合TEM提供了一种敏感的空间离散技术，用于评估外周动脉功能受损的恶性后遗症的治疗对策的机制基础和疗效。
Peripheral artery disease (PAD) in the lower limb compromises oxygen supply due to arterial occlusion. Ischemic skeletal muscle is accompanied by capillary structural deformation. Therefore, using novel microscopy techniques, we tested the hypothesis that endothelial cell swelling temporally and quantitatively corresponds to enhanced microvascular permeability. Hindlimb ischemia was created in male Wistar rat’s by iliac artery ligation (AL). The tibialis anterior (TA) muscle microcirculation was imaged using intravenously infused rhodamine B isothiocyanate dextran fluorescent dye via two-photon laser scanning microscopy (TPLSM) and dye extravasation at 3 and 7 days post-AL quantified to assess microvascular permeability. The TA microvascular endothelial ultrastructure was analyzed by transmission electron microscopy (TEM). Compared with control (0.40 ± 0.15 μm3× 106), using TPLSM, the volumetrically determined interstitial leakage of fluorescent dye measured at 3 (3.0 ± 0.40 μm3× 106) and 7 (2.5 ± 0.8 μm3× 106) days was increased (bothP< 0.05). Capillary wall thickness was also elevated at 3 (0.21 ± 0.06 μm) and 7 (0.21 ± 0.08 μm) days versus control (0.11 ± 0.03 μm, bothP< 0.05). Capillary endothelial cell swelling was temporally and quantitatively associated with elevated vascular permeability in the AL model of PAD but these changes occurred in the absence of elevations in protein levels of vascular endothelial growth factor (VEGF) its receptor (VEGFR2 which decreased by AL-7 day) or matrix metalloproteinase. The temporal coherence of endothelial cell swelling and increased vascular permeability supports a common upstream mediator. TPLSM, in combination with TEM, provides a sensitive and spatially discrete technique to assess the mechanistic bases for, and efficacy of, therapeutic countermeasures to the pernicious sequelae of compromised peripheral arterial function.