| 卢琼 苏华斌 王蕾 吴和平.高糖刺激下PKC/NADPH氧化应激途径对内皮细胞活性氧生成的影响及其机制探讨[J].,2015,15(30):5843-5845 |
| 高糖刺激下PKC/NADPH氧化应激途径对内皮细胞活性氧生成的影响及其机制探讨 |
| Effects of PKC/NADPH Oxidase on ROS in HUVECs Culturedwith High Glucose |
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| DOI： |
| 中文关键词: NADPH 氧化酶 活性氧 葡萄糖 蛋白激酶C p47phox |
| 英文关键词: NADPH oxidase Reactive oxygen species Glucose Protein kinase c p47phox |
| 基金项目:湖南省教育厅科研基金项目（13C739）；湖南医药学院校级科研基金项目（2011KY02） |
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| 中文摘要: |
| 目的：探讨高糖刺激下PKC/NADPH 氧化应激途径对内皮细胞活性氧生成的影响。方法：实验分为正常对照组、NaOH对照
组、DMSO 对照组、20 mM葡萄糖处理4 小时组（高糖组）、1 mol佛波酯预处理0.5 小时再加入20 mM葡萄糖处理组（佛波酯组）
和4 mol 金丝桃素预处理0.5 小时再加入20 mM 葡萄糖处理组（金丝桃素组）；Ⅱ型胶原酶消化法分离人脐静脉内皮细胞；流式
细胞术检测细胞内活性氧；免疫荧光检测内皮细胞Ⅷ因子和NADPH 氧化酶亚基p47phox 定位。结果：①与正常对照组相比，高糖
组细胞内活性氧增高（P<0.05，n=3），与正常对照组和高糖组相比，佛波酯组HUVECs内活性氧的产生显著增加（P<0.05，n=3），与
正常对照组和高糖组相比，金丝桃素组HUVECs 内活性氧的产生明显减少（P<0.05，n=3）；②正常对照组和金丝桃素组中细胞
NADPH 氧化酶亚基p47phox 主要位于胞浆，而佛波酯组和高糖组的NADPH 氧化酶亚基p47phox位于胞膜。结论：高糖通过
PKC 信号通路调节内皮细胞NADPH 氧化酶亚基p47phox 的移位从而增加细胞内活性氧的生成。 |
| 英文摘要: |
| Objective:To investigate effects of PKC/NADPH oxidase on reactive oxygen species (ROS) in Human umbilical vein
endothelial cells (HUVECs) cultured with high glucose.Methods:The experiments were divided into six groups, including control group,
NaOH control group, DMSO control group, group treated with glucose (20 mM) for 4 hours (high-glucose group), group which was
pretreated phorbol ester (1 mol) for half an hour then was stimulated by glucose (20 mM) for 4 hours (phorbol ester group) and group
which was pretreated hypericin (4 mol) for half an hour then was stimulated by glucose (20 mM) for 4 hours(hypericin group); Human
umbilical vein endothelial cells was isolated by type Ⅱ collegease; flow cytometry was used to detect intracellular ROS production;
immunofluorescence was used to investigate factor Ⅷ and p47phox location in HUVECs.Results:① Compared with control group,
intracellular ROS of high-glucose group were significantly increased （P<0.05, n=3）, Compared with control group and, high-glucose
group, intracellular ROS of phorbol ester group were significantly increased （P<0.05, n=3）, Compared with control group and,
high-glucose group, intracellular ROS of hypericin group were significantly decreased （P<0.05, n=3）; ② p47phox mostly located
cytoplasm for control group and hypericin group, p47phox mostly located cell membrane for high-glucose group and phorbol ester group.Conclusion:PKC/NADPH oxidase can regulate ROS generation in HUVECs induced with high glucose by translocation of p47phox. |
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