天津医药

天津医药 ›› 2017, Vol. 45 ›› Issue (9): 907-911.doi: 10.11958/20170534

• 细胞与分子生物学 • 上一篇    下一篇

NADPH 对人脐静脉内皮细胞缺糖缺氧复灌损伤早期的保护作用

宋琪 1，翁晓芬 1，董婷 1，许莉敏 1，谢燕 1，秦正红 2   

  1. 1 苏州大学附属第一医院（邮编 215123）；2 苏州大学药学院神经疾病与衰老实验室
  • 收稿日期:2017-05-03 修回日期:2017-08-23 出版日期:2017-09-15 发布日期:2017-09-25
  • 通讯作者: 谢燕 E-mail:xieyan005@163.com
  • 基金资助:
    TIGAR在脑缺血时表达的调节及对神经损伤的影响

The early protective effect of NADPH on OGD/R injury of human umbilical vein endothelial cells

SONG Qi1, WENG Xiao-fen1, DONG Ting1, XU Li-min1, XIE Yan1, QIN Zheng-hong2   

  • Received:2017-05-03 Revised:2017-08-23 Published:2017-09-15 Online:2017-09-25

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摘要： 摘要：目的 探讨还原型辅酶Ⅱ（NADPH）对人脐静脉内皮细胞（HUVECs）缺糖缺氧复灌（OGD/R）损伤早期的保 护作用及其对 occludin、基质金属蛋白酶（MMP）9 蛋白表达的影响。方法 体外培养 HUVECs 系，建立稳定的 HUVECs 缺糖缺氧复灌损伤模型，实验分 3 组：正常对照组、缺糖缺氧复灌组、缺糖缺氧复灌+NADPH 组。细胞增殖 试剂盒（CCK-8 法）检测 HUVECs 活力；细胞毒性试剂盒检测 HUVECs 内乳酸脱氢酶（LDH）释放；倒置显微镜观察 HUVECs 形态变化；比色法检测 HUVECs 裂解液中超氧化物歧化酶（SOD）活力及丙二醛（MDA）含量；Western Blot 法检测各组 occludin、MMP9 蛋白的表达。结果 与缺糖缺氧复灌组比，NADPH 处理可提高缺糖缺氧复灌损伤后 HUVECs 的存活率（P＜0.05），减少损伤后 HUVECs 中 LDH 释放（P＜0.05），促进 HUVECs 形态的恢复，减少 MDA 生成（P＜0.05），提高 SOD 活力（P＜0.05）；缺糖缺氧复灌处理后，给予 NADPH 处理可抑制 MMP9 水平上调（P＜0.05），促进 occludin 水平恢复（P＜0.05）。结论 NADPH 对 HUVECs 具有保护作用，其保护作用与抑制氧化应激反应，调节 MMP9、occludin 蛋白的表达有关。

关键词: NADPH, 内皮细胞, 脐静脉, 氧化性应激, 基质金属蛋白酶 9, occludin, 缺糖缺氧复灌

Abstract: Abstract: Objective To study the early protective effect of NADPH on human umbilical vein endothelial cells (HUVECs) and the expression of occludin and MMP9 induced by oxygen glucose deprivation and reoxygenation (OGD/R). Methods HUVECs were cultured and divided into blank control group, OGD/R group and OGD/R+NADPH 20 μmol/L group. The proliferation of HUVECs after treatment was detected by CCK- 8 assay. The cytotoxicity was detected by LDH release assay. The morphological changes of HUVECs were observed by inverted microscope. Superoxide dismutase (SOD MDA) activity and malondialdehyde (MDA) were detected by commercially available kit. The expressions of occludin and MMP9 were detected by Western blot assay. Results Compared with the OGD/R, NADPH enhanced the cell viability significantly (P＜0.05), reduced the release of LDH (P＜0.05), promoted the maintance of HUVECs morphology, reduced MDA generation (P＜0.05) and increased SOD activity (P＜0.05). Following OGD/R，the treatment of NADPH can inhibit MMP9 level (P＜0.05) and promote the recovery of occludin level (P＜0.05). Conclusion NADPH can protect HUVECs from the damage induced by OGD/R by reducing oxidative stress and regulating the expressions of MMP9 and occludin.

Key words: NADPH, endothelial cells, umbilical veins, oxidative stress, matrix metalloproteinase 9, occludin, oxygen glucose deprivation and reoxygenation