氯化钴诱导人肝窦内皮细胞缺氧模型特点

肝脏 ›› 2018, Vol. 23 ›› Issue (3): 217-220.

氯化钴诱导人肝窦内皮细胞缺氧模型特点

张风, 王清兰, 刘成海, 陶艳艳

201203 上海中医药大学附属曙光医院肝病研究所(张风,刘成海,陶艳艳);上海高校中医内科学E-研究院(王清兰)

收稿日期:2017-12-04 发布日期:2020-04-30
通讯作者: 陶艳艳,Email:taoyanyan1023@126.com
基金资助:
国家自然科学基金(81573810);上海市自然科学基金(16ZR1437800)

Characteristics of hypoxia model induced by cobalt chloride in human hepatic sinusoidal endothelial cells

ZHANG Feng, WANG Qing-lan, LIU Cheng-hai, TAO Yan-yan

Institute of Liver Diseases, Shuguang Hospital affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 201203;E-Institute of Traditional Chinese Medicine Internal Medicine, Shanghai Municipal Education Commission, Shanghai 201203;Shanghai Key Laboratory of Traditional Chinese Clinical Medicine, Shanghai 201203

Received:2017-12-04 Published:2020-04-30
Contact: TAO Yan-yan,Email:taoyanyan1023@126.com

摘要/Abstract

摘要： 目的观察氯化钴(CoCl₂)诱导人肝窦内皮细胞(HHSEC)缺氧模型特点。方法不同浓度CoCl₂(0～800 μmol/L)与HHSEC细胞孵育24 h,CCK8法检测细胞活力;50～200 μmol/L CoCl₂与HHSEC细胞孵育,Transwell实验检测细胞迁移;Matrigel管腔生成实验检测细胞管腔形成情况;蛋白质免疫检测细胞HIF-1α、vWF、VEGF蛋白表达;免疫荧光染色观察HIF-1α核转位。结果与对照组比较,50～200 μmol/L CoCl₂组HHSEC细胞活力明显增加,细胞迁移到膜下的数量显著增多,并且形成丰富而密闭的管状结构。50 μmol/L、200 μmol/L CoCl₂促进HHSEC中缺氧诱导因子1α(HIF-1α)、第八因子相关抗原(vWF)、血管内皮生长因子(VEGF)表达,其中200 μmol/L CoCl₂组HIF-1α、vWF表达较50 μmol/L CoCl₂组高。结论 50 μmol/L、200 μmol/L低浓度CoCl₂可诱导人肝窦内皮细胞缺氧、血管新生,其作用机制与促进HIF-1α表达及核转位相关。

关键词: 氯化钴, 人肝窦内皮细胞, 缺氧, 缺氧诱导因子-1α

Abstract: Objective To investigate the characteristics of hypoxia model induced by cobalt chloride (CoCl₂) in human hepatic sinusoidal endothelial cells (HHSEC).Methods HHSEC were incubated with different concentrations of CoCl₂ (0～800 μM) for 24 h, in which cell viability was measured with counting kit-8 (CCK8) assay. Cell migration was detected by transwell assay after stimulating HHSEC with 50～200 μM CoCl₂. Matrigel lumen formation assay was used to detect cell lumen formation. Expressions of cell hypoxia inducible factor -1α (HIF-1α), von Willebrand factor (vWF) and vascular endothelial growth factor (VEGF) protein were detected by western blot. HIF-1α nuclear translocation was observed by immunofluorescence staining. Results Compared with control group, cell viability and cell migration of HHSEC were significantly increased, with a rich and airtight tubular structure formed in low concentration of 50～200 μM CoCl₂ group. Expressions of HIF-1α, vWF and VEGF in HHSEC were increased in 50 μM and 200 μM CoCl₂ group than those in the control group. Furthermore, expressions of HIF-1α and vWF in 200μM CoCl₂ group were higher than those in 50 μM CoCl₂ group.Conclusion Low levels of 50 μM and 200 μM CoCl₂ may induce HHSEC hypoxia and angiogenesis through promoting the expression and nuclear translocation of HIF-1α.

Key words: Cobalt chloride, Human hepatic sinusoidal endothelial cells, Hypoxia, Hypoxia-inducible factor-1α

张风, 王清兰, 刘成海, 陶艳艳. 氯化钴诱导人肝窦内皮细胞缺氧模型特点[J]. 肝脏, 2018, 23(3): 217-220.

ZHANG Feng, WANG Qing-lan, LIU Cheng-hai, TAO Yan-yan. Characteristics of hypoxia model induced by cobalt chloride in human hepatic sinusoidal endothelial cells[J]. Chinese Hepatolgy, 2018, 23(3): 217-220.