番茄红素对高糖所致内皮祖细胞线粒体氧化损伤和功能的影响

EFFECTS OF LYCOPENE ON OXIDATIVE DAMAGE AND FUNCTION OF ENDOTHELIAL PROGENITOR CELLS MITOCHONDRIA EXPOSED TO HIGH GLUCOSE

目的 观察高糖对大鼠骨髓内皮祖细胞（EPCs）线粒体氧化损伤情况,并探讨番茄红素（Lyc）对高糖所致的EPCs线粒体氧化损伤的保护作用。方法 密度梯度离心法获取骨髓单个核细胞,体外诱导分化后,用FITC标记的荆豆凝集素Ⅰ（FITC-UEA-I）和Dil标记的乙酰化低密度脂蛋白（Dil-ac-LDL）双染色鉴定为EPCs。分离纯化的细胞随机分为6个组：（1）正常对照组,（2）甘露醇对照组,（3）高糖对照组,（4~6）为Lyc低、中、高剂量组,分别用含终浓度为10、30、50μg/ml的Lyc加入33 mmol/L高糖培养基。培养24 h后,检测线粒体氧化损伤和功能相关指标,并分析不同组间指标变化。结果 经不同浓度Lyc共培养后,高糖对照组EPCs线粒体呼吸酶复合物Ⅰ、Ⅱ、Ⅲ、Ⅳ活性和线粒体膜电位均低于正常对照组（P〈0.05）,高糖对照组EPCs线粒体ROS和8-OHdG均高于正常对照组（均P〈0.05）。Lyc干预后,低、中、高剂量组EPCs线粒体呼吸酶复合物Ⅰ、Ⅱ、Ⅲ、Ⅳ活性均高于高糖对照组活性（均P〈0.05）。Lyc低、中、高剂量组EPCs线粒体ROS与8-OHdG均低于高糖对照组（均P〈0.05）。结论 高糖可以诱导EPCs线粒体氧化损伤和功能下降。Lyc对糖所致EPCs线粒体氧化损伤及线粒体复合物活性的降低有较好的保护作用。

Objective To study the oxidative damage of endothelial progenitor cells＇ mitochondria induced by high concentration of glucose and investigate the protective effects of lycopene （Lyc）. Methods Mononuclear cells were isolated from endothelial progenitor cells of Sprague-Dawley rats＇ bone marrow by Ficoll density gradient centrifugation. The cells were plated on fibronectin-coated culture dishes. After being induced to differentiation, the EPCs were identified through fluorescein labeled Ulex Europaeus Agglutinin I （FITC-UEA-I） and Dil labeled acetylated low density lipoprotein （Dil-ac-LDL） double staining. The identified EPCs were randomly divided into 6 groups, i e. control group, mannitol group, high glucose group, high glucose＋Lyc low dose group, high glucose＋Lyc middle dose group and high glucose＋Lyc high dose group. EPCs in the Lyc low, middle and high dose groups were incubated with 10, 30, and 50 μg/ml Lyc plus high glucose respectively. After 24h incubation, EPCs mitocbondria were isolated and used to detect the related indexes of oxidative damages. Results The activity of mitoehondrial atmungsferment complex I, Ⅱ, Ⅲ, IV and mitochondrial transmembrane potential （MMP） in the high glucose group were significantly lower than those in the control group（P〈0.05）. The content of reactive oxygen specios （ROS） and 8-hydroxy-2＇ -deoxyguanosine （8-OHdG） in the h!gh glucose group were significantly higher than those in the control group （P〈0.05）. After intervention With Lyc, the activity of mitochondrial atmungsferment complex I, Ⅱ , Ⅲ, IV and MMP were significantly higher than those in high glucose group （P〈0.05）. The content of ROS and 8-OHdG in the Lyc low, middle and high dose groups were lower than those in the high glucose group （P〈0.05）.Conclusion High glucose can induce mitochondrial oxidative dysfunction in EPCs, while Lyc shows protective effect on EPCs mitochondrial complex.