中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (20): 3723-3727.doi: 10.3969/j.issn.1673-8225.2011.20.027

• 组织构建与中医药 tissue construction and traditional Chinese medicine • 上一篇    下一篇

灯盏花乙素干预缺血再灌注损伤人脐静脉内皮细胞血管内皮生长因子与蛋白激酶Cε的表达

李  琳1，刘  华2，蔡红雁1，杨为民3，郭  涛1   

  1. 昆明医学院第一附属医院，1心内科，2检验科，云南省昆明市 650032；3昆明医学院云南省天然药物药理重点实验室，云南省昆明市 650032
  • 收稿日期:2011-02-14 修回日期:2011-03-19 出版日期:2011-05-14 发布日期:2011-05-14
  • 通讯作者: 郭涛，硕士，教授，主任医师，昆明医学院第一附属医院心内科，云南省昆明市 650032 guotao20@ hotmail.com 通讯作者：杨为民，博士，研究员，昆明医学院云南省天然药物药理重点实验室，云南省昆明市650032 ywmbessie@ yeah.net
  • 作者简介:李琳☆，女，1973年生，四川省荣县人，汉族，昆明医学院在读博士，主治医师，主要从事高血压、冠心病基础及临床研究。 lilinkm@yahoo. com.cn
  • 基金资助:

    国家自然科学基金资助项目(30960450)，灯盏花乙素拮抗缺血-再灌注诱导的微小血管内皮损伤。

Effects of scutellarin on vascular endothelial growth factor and protein kinase Cε expression in human umbilical vein endothelial cells after ischemia-reperfusion injury

Li Lin1, Liu Hua2, Cai Hong-yan1, Yang Wei-min3, Guo Tao1   

  1. 1Department of Cardiology, 2Clinical Laboratory, First Affiliated Hospital of Kunming Medical University, Kunming  650032, Yunnan Province, China;    3Yunnan Pharmacology Lab of Natural Product, Kunming Medical University, Kunming  650032, Yunnan Province, China
  • Received:2011-02-14 Revised:2011-03-19 Online:2011-05-14 Published:2011-05-14
  • Contact: Guo Tao, Master, Professor, Chief physician, Department of Cardiology, First Affiliated Hospital of Kunming Medical University, Kunming 650032, Yunnan Province, China guotao20@hotmail. com Correspondence to: Yang Wei-min, Doctor, Investigator, Yunnan Pharmacology Lab of Natural Product, Kunming Medical University, Kunming 650032, Yunnan Province, China ywmbessie@yeah. net
  • About author:Li Lin☆, Studying for doctorate, Attending physician, Department of Cardiology, 2Clinical Laboratory, First Affiliated Hospital of Kunming Medical University, Kunming 650032, Yunnan Province, China lilinkm@yahoo.com. cn
  • Supported by:

    the National Natural Science Foundation of China, No. 30960450*

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摘要：

背景：有关灯盏花乙素的研究多集中在神经细胞、神经元及平滑肌细胞上，但对血管内皮细胞的作用研究尚少。
目的：观察灯盏花乙素预处理后对人脐静脉内皮细胞缺血再灌注损伤后血管内皮生长因子与蛋白激酶Cε表达的影响。
方法：体外培养的人脐静脉内皮细胞，分别设立对照组，模型组和及灯盏花乙素高、中、低剂量组。将人脐静脉内皮细胞予缺氧缺糖3 h/复氧糖5或24 h；灯盏花乙素高、中、低剂量组分别加1×10-5，1×10-6，1×10-7 mol/L灯盏花乙素孵育30 min，然后予缺血再灌注处理。实验结束后取细胞裂解液用Western blot检测血管内皮生长因子、蛋白激酶Cε的蛋白表达。
结果与结论：缺血3 h再灌注5及24 h，较对照组，模型组血管内皮生长因子的表达降低，细胞颗粒部分蛋白激酶Cε的表达明显增加。灯盏花乙素能促进缺血3 h再灌注5 h人脐静脉内皮细胞血管内皮生长因子的表达，尤其以灯盏花乙素高剂量和中剂量组明显，但对蛋白激酶Cε的表达没有影响。

关键词: 灯盏花乙素, 血管内皮细胞, 缺血/再灌注, 血管内皮生长因子, 蛋白激酶Cε

Abstract:

BACKGROUND: The effects of scutellatin on ischemia-reperfusion injury to endothelial cells in vitro have not been elucidated.
OBJECTIVE: To investigate the effect of scutellatin on human umbilical vein endothelial cells (HUVECs) after ischemia-reperfusion and to study whether this effect is mediated by vascular endothelial growth factor (VEGF) and protein kinase Cε (PKCε).
Methods: Cultured HUVECs, exposed to oxygen and glucose deprivation followed by reperfusion, were used as an in vitro model of ischemia-reperfusion. After incubated with scutellarin in a low (1×10-7 mol/L), middle (1×10-6 mol/L) and high (1×10-5 mol/L) dose for 30 minutes, cells were treated by ischemia 3 hours/reperfusion 5 to 24 hours. VEGF protein expression was evaluated by Western blotting. The translocation of PKCε was assayed by Western blotting.
RESULTS AND CONCLUSION: Ischemia 3 hours/reperfusion 5 hours injury significantly decreased the expression of VEGF, and scutellatin in the high and middle dose significantly increased VEGF expression (P < 0.05). Under ischemia 3 hours/reperfusion 24 hours condition, VEGF expression was not changed and scutellatin failed to further increase the expression of VEGF. Compared with the control group, there were significant increases of PKCε expression in particulate fractions in ischemia-reperfusion group and scutellatin pretreatment groups (P < 0.01). Compared with ischemia-reperfusion group, there was no further increase of PKCε in scutellatin groups. Scutellarin pretreatment on vascular endothelial cells after ischemia-reperfusion injury may have a protective effect, and the mechanism may be related to the early increase of VEGF.

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