藏红花素对人脐静脉内皮细胞增殖及细胞外调节蛋白激酶信号通路的影响

doi:10.16098/j.issn.0529-1356.2017.03.006

解剖学报 ›› 2017, Vol. 48 ›› Issue (3): 282-286.doi: 10.16098/j.issn.0529-1356.2017.03.006

藏红花素对人脐静脉内皮细胞增殖及细胞外调节蛋白激酶信号通路的影响

赵玉娇¹ 春花² 李鑫磊¹ 吴梦楠¹杨慧科^1*

1. 哈尔滨医科大学人体解剖学教研室，哈尔滨 150081; 2. 西藏藏医学院基础部现代医药教研室，拉萨 850000

收稿日期:2016-11-18 修回日期:2017-01-04 出版日期:2017-06-06 发布日期:2017-09-19
通讯作者: 杨慧科 E-mail:huikeyang@aliyun.com
基金资助:
藏红花素通过调节离子通道KCa3.1、KCa2.3改善糖尿病血管内皮功能的作用研究;KCa3.1在淋巴管内皮的表达和角膜淋巴管新生中的作用研究;IKCa1通道在VEGF诱导脐静脉内皮细胞增殖中的调节作用

Effects of crocin on the cell proliferation and extracellular regulated protein kinases pathway in human umbilical vein endothelial cells

ZHAO Yu-jiao¹ CHUN Hua² LI Xin-lei¹ WU Meng-nan¹ YANG Hui-ke^1*

1. Department of Anatomy, Harbin Medical University, Harbin 150081, China; 2. Department of Modern Medicine, Tibet Traditional Medical College, Lhasa 850000, China

Received:2016-11-18 Revised:2017-01-04 Online:2017-06-06 Published:2017-09-19
Contact: YANG Hui-ke E-mail:huikeyang@aliyun.com

摘要/Abstract

摘要：

目的观察藏红花素(crocin)对人脐静脉内皮细胞（HUVECs）体外增殖、细胞外调节蛋白激酶（ERK）1/2表达及活性的影响。方法用藏红花素及MAPK/ERK激酶（MEK）抑制剂PD98059分别处理HUVECs，EdU细胞增殖法检测细胞体外增殖活力，Western blotting法检测crocin对磷酸化细胞外调节蛋白激酶1/2（ERK1/2）和总ERK1/2表达的影响，激光扫描共焦显微镜检测细胞内Ca²⁺浓度。结果在1μmol/L和10μmol/L浓度水平，藏红花素可明显促进细胞的增殖能力，提高磷酸化ERK1/2和总ERK1/2蛋白的表达水平，并增加细胞内Ca²⁺浓度；给予MEK抑制剂PD98059后，细胞的增殖能力下降，磷酸化ERK1/2和总ERK1/2的表达减弱，细胞内Ca²⁺浓度降低。结论藏红花素通过活化ERK1/2信号途径，提高细胞内Ca²⁺浓度，促进人脐静脉内皮细胞的体外增殖能力。

关键词: 藏红花素, 人脐静脉内皮细胞, 增殖, 细胞外调节蛋白激酶, 免疫印迹法

Abstract:

Objective To investigate the effects of crocin on the proliferation and ERK1/2 pathway in human umbilical vein endothelial cells (HUVECs) in vitro. Methods In this study, HUVECs were treated by crocin with or without the MAPK/ERK kinase(MEK) inhibitor PD98059, and the proliferation of cells was measured by EdU assay. The expressions of phospho-ERK1/2 (p-ERK1/2) and total-ERK1/2 (t-ERK1/2) were analyzed by Western blotting. In addition, the concentration of intracellular Ca²⁺ was detected by confocal microscopy to evaluate the underlying effect of crocin on the cell proliferation. Results EdU assay showed that crocin stimulated the cell proliferation effectively, especially at the concentration of 1μmol/L and 10μmol/L. Western blotting results revealed that crocin improved the expression of p-ERK1/2 and t-ERK1/2 in HUVECs. The increase of the intracellular Ca²⁺ concentration by crocin at 1μmol/L and 10μmol/L concentration was importantly declared using Fluo3AM labeling. After the inhibitor of MEK PD98059 was given, decreased the level of cell proliferation, and the p-ERK1/2 and t-ERK1/2 expression and the intracellular Ca²⁺ concentration had decreased as well. Conclusion Crocin has effect on the proliferation of HUVECs and increase the intracellular Ca²⁺ through ERK1/2 signal pathway.

Key words: Crocin, Human umbilical vein endothelial cell, Proliferation, Extracellular signal-regulated kinase, Western blotting

赵玉娇春花李鑫磊吴梦楠杨慧科. 藏红花素对人脐静脉内皮细胞增殖及细胞外调节蛋白激酶信号通路的影响[J]. 解剖学报, 2017, 48(3): 282-286.

ZHAO Yu-jiao CHUN Hua LI Xin-lei WU Meng-nan YANG Hui-ke. Effects of crocin on the cell proliferation and extracellular regulated protein kinases pathway in human umbilical vein endothelial cells[J]. Acta Anatomica Sinica, 2017, 48(3): 282-286.

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藏红花素对人脐静脉内皮细胞增殖及细胞外调节蛋白激酶信号通路的影响

Effects of crocin on the cell proliferation and extracellular regulated protein kinases pathway in human umbilical vein endothelial cells

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