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

作　　者：赵玉娇[1] 春花[2] 李鑫磊[1] 吴梦楠[1] 杨慧科[1] ZHAO Yu-jiao CHUN Hua LI Xin-lei WU Meng-nan YANG Hui-ke(Department of Anatomy, Harbin Medical University, Harbin 150081, China Department of Modern Medicine, Tibet Traditional Medical College, Lhasa 850000, China)

机构地区：[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

出　　处：《解剖学报》2017年第3期282-286,共5页Acta Anatomica Sinica

基　　金：国家自然科学基金(81460660);黑龙江省自然科学基金(QC2013C017);黑龙江省卫生厅科研课题(2011-212)

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,Ed U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Ca^(2+)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Ca^(2+)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Ca^(2+)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Ca^(2+)浓度,促进人脐静脉内皮细胞的体外增殖能力。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 erocin 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 confoeal 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 ll^mol/L and 101~mol/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 l lxmol/L and lOixmol/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 intraeellular 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.

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 

分 类 号：R34[医药卫生—基础医学]