D-4F在七氟烷处理对急性高糖血症小鼠血管保护中的作用及机制

目的: 观察apoA-1拟似物D-4F对七氟烷麻醉急性高糖血症小鼠的血管保护效应。方法: 实验一，雄性 C57BL/6J小鼠随机分4组（n=8）：对照组（CON组）、 D-4F组、急性高血糖组（AHG组）、 AHG+D-4F组，各组再进行吸入1.71%七氟烷(SEV)，检测各组血糖、主动脉血管环舒张功能、NO水平，Western blot检测内皮型一氧化氮合酶（eNOS）、磷酸化的eNOS（p-eNOS）、小凹蛋白（Cav-1）水平；实验二，人脐静脉血管内皮细胞（HUVEC）分别用5.5、20.0 mmol/L 的葡萄糖培养24 h，4 μg/mL 布雷德菌素A (BFA)和0.5 μg/mL D-4F孵育1 h，再进行SEV处理，检测ROS水平。结果: AHG组显示急性高血糖时血管的舒张能力下降，主动脉产生NO减少，Cav-1蛋白表达明显增强，eNOS活性下降。D-4F组显示加入D-4F并不能提高急性高血糖时的血管舒张能力，AHG+D-4F组显示给予SEV刺激后D-4F对血管的舒张有改善作用（P<0.01)，Cav-1蛋白表达下降，eNOS活性增强，NO生成增多。细胞实验显示SEV刺激下，HG+D-4F组ROS的生成下降，D-4F该功能并可被BFA所阻断。结论: D-4F可恢复SEV对急性高糖血症的血管保护功能，其机制可能与D-4F降低细胞内脂质筏区域Cav-1的耦合能力，增强脂质筏里eNOS的磷酸化，NO生成增多，ROS生成减少，减轻急性高血糖的血管损害有关。

D-4F enhances sevoflurane-induced vascular protection during acute hyperglycemia

AIM: To investigate the effects of sevoflurane-induced vascular protection by apolipoprotein A-1 mimetic D-4F during acute hyperglycemia. METHODS:Thrty-two male C57BL/6J mice were randomly assigned to 4 groups (n=8). Control group(CON group), D-4Fgroup, acute hyperglycemia group(AHG group), dextrose 2 g/kg (intraperitoneal injection); AHG+D-4F group. Blood glucose, isometric aortic tension, nitric oxide (NO) were measured in mice in the absence or presence of sevoflurane（1.71%）. Expressions of eNOS，p-eNOS，Cav-1 in arterioles were determined by Western blot.Superoxide generation species (ROS) were assessed in human umbilical vein endothelial cells (HUVEC) cultured in 5.5 or 20.0 mmol/L glucose with isoflurane (0.5 mmol/L) in the presence or absence of brefeldin A (BFA, 4 μg/mL) or D-4F (0.5 μg/mL). RESULTS: Vasodilation, phosphorylations, NO production of eNOS were decreased while Cav-1 expression was increased during acute hyperglycemia. D-4F increased endothelium-dependent vasodilation (P<0.01) while decreased Cav-1 expression and ROS production in HG+D-4F group after sevoflurane-induction.CONCLUSION: Polipoprotein A-1 mimetic D-4F can enhance sevoflurane-induced vascular protection, decrease Cav-1 expression and ROS production while increase NO production during acute hyperglycemia.