Low molecular-weight fucoidan protects against hindlimb ischemic injury in type 2 diabetic mice through enhancing endothelial nitric oxide synthase phosphorylation
Low molecular-weight fucoidan protects against hindlimb ischemic injury in type 2 diabetic mice through enhancing endothelial nitric oxide synthase phosphorylation
复制标题
低分子量岩藻依聚糖通过增强内皮一氧化氮合酶磷酸化保护 2 型糖尿病小鼠后肢缺血性损伤
DOI：
10.1111/1753-0407.12667
复制
发表时间：
2018-11-01
影响因子：
4.5
通讯作者：
Luo, Dali
中科院分区：
文献类型：
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作者：
Liu, Tiantian;Wang, Zhiqiang;Luo, Dali
背景：糖尿病（DM）并发症与缺血性损伤有关。血管生成是糖尿病足的一种治疗策略。本研究的目的是探讨低分子量岩藻糖聚糖（LMWF）在糖尿病外周动脉疾病（PAD）中可能的血管生成作用。方法：糖尿病db/db小鼠和年龄匹配的C57BL/6小鼠行股动脉结扎后，分别给予低分子肝素（30、60、80 mg/kg/d, p.o）或西洛他唑（30 mg/kg/d, p.o）治疗6周。测量后肢内皮依赖性血管舒张和血流量。对腓肠肌组织中CD34、血管内皮生长因子（VEGF）、eNOS和炎症因子进行组织学和western blot分析。在人脐静脉内皮细胞（HUVEC）中的作用得到证实。结果：结扎后的糖尿病小鼠出现后肢溃疡、肉瘤和坏死，炎症因子表达增加，VEGF和eNOS磷酸化水平降低。LMWF治疗可显著改善糖尿病PAD小鼠足部病变，抑制炎症因子表达，并通过促进内皮依赖性血管扩张和血管重建改善足底灌注。在高糖处理的HUVEC中，LMWF （40 μ g/mL）逆转了内皮细胞的增殖、迁移和小管形成，促进了eNOS磷酸化和VEGF表达，而用eNOS拮抗剂n -g -硝基-L-精氨酸甲基酯（100 μ mol/L）预处理HUVEC可明显抑制LMWF的作用。结论：本研究表明，LMWF减轻后肢缺血损伤，至少部分是通过促进eNOS磷酸化、一氧化氮生成和VEGF表达，从而增强缺血区域的血管生成。
Background: Diabetes mellitus (DM) complications are associated with ischemic injury. Angiogenesis is a therapeutic strategy for diabetic foot. The aim of this study was to investigate the possible angiogenic effect of low molecular weight fucoidan (LMWF) in diabetic peripheral arterial disease (PAD).Methods: Diabetic db/db mice and age-matched C57BL/6 mice underwent femoral artery ligation followed by LMWF (30, 60, 80 mg/kg per day, p.o.) or cilostazol (30 mg/kg/day, p.o.) treatment for 6 weeks. Endothelium-dependent vasodilation and blood flow of the hindlimb were measured. Histological and western blot analyses of CD34, vascular endothelial growth factor (VEGF), eNOS, and inflammatory factors in the gastrocnemius were performed. The effects of LMWF were confirmed in human umbilical vein endothelial cells (HUVEC).Results: Diabetic mice with ligation exhibited hindlimb ulceration, hydrosarca, and necrosis, increased expression of inflammatory factors, and decreased levels of VEGF and eNOS phosphorylation. Treatment with LMWF markedly ameliorated foot lesions, suppressed expression of inflammatory factors, and improved plantar perfusion by promoting endothelium-dependent vasodilation and revascularization in diabetic PAD mice. In high-glucose treated HUVEC, LMWF (40 mu g/mL) reversed blunted endothelial cell proliferation, migration, and tube formation, and promoted eNOS phosphorylation and VEGF expression, whereas HUVEC pretreatment with 100 mu mol/L N-G-nitro-L-arginine methyl ester, an eNOS antagonist, markedly inhibited the effects of LMWF.Conclusion: This study demonstrates that LMWF alleviates hindlimb ischemic damage, at least in part by promoting eNOS phosphorylation, nitric oxide production, and VEGF expression, resulting in enhanced angiogenesis in the ischemic region.