烟酸姜黄素酯对低剪切应力诱导人脐静脉内皮细胞凋亡的影响及相关机制

doi:10.12092/j.issn.1009-2501.2018.08.003

中国临床药理学与治疗学 ›› 2018, Vol. 23 ›› Issue (8): 855-861.doi: 10.12092/j.issn.1009-2501.2018.08.003

烟酸姜黄素酯对低剪切应力诱导人脐静脉内皮细胞凋亡的影响及相关机制

刘静¹，王媚媚²，刘录山³，贺卫和⁴，庹勤慧⁴，文红艳⁴

¹湖南中医药大学中西医结合学院，长沙 410208，湖南；²南华大学附属南华医院儿科，衡阳 421002，湖南；³南华大学心血管研究所，衡阳 421001，湖南；4湖南中医药大学医学院，长沙 410208，湖南

收稿日期:2018-04-20 修回日期:2018-05-24 出版日期:2018-08-26 发布日期:2018-08-28
通讯作者: 文红艳，女，主任医师，教授，硕导，研究方向：中医药防治心脑血管疾病。 Tel:15111275189 E-mail:340681499@qq.com
作者简介:刘静，女，硕士研究生，研究方向：中医药防治心脑血管疾病。 Tel:18774050315 E-mail:570429747@qq.com
基金资助:
国家自然科学基金（81673722）；湖南省教育厅重点项目（201515A）；湖南省自然科学基金资助项目(2015JJ4097)；湖南省大学生研究性学习和创新性实验计划项目（2017286）

Effects and mechanisms of nicotinic acid curcumin on apoptosis induced by low shear stress in human umbilical vein endothelial cells

LIU Jing¹, WANG Meimei², LIU Lushan³, HE Weihe⁴, TUO Qinhui⁴, WEN Hongyan⁴

¹College of Chinese and Western Medicine, Hunan University of Chinese Medicine, Changsha 410208, Hunan, China;² Department of Pediatric, Affiliated Nanhua Hospital, University of South China, Hengyang 421002, Hunan, China; ³Institute of Cardiovascular Disease, University of South China, Hengyang 421001, Hunan, China; ⁴Medical School, Hunan University of Chinese Medicine, Changsha 410208, Hunan, China

Received:2018-04-20 Revised:2018-05-24 Online:2018-08-26 Published:2018-08-28

摘要/Abstract

摘要：

目的: 探讨烟酸姜黄素酯（curcumin nicotinate，CN）对低剪切应力（low shear stress, LSS）诱导人脐静脉内皮细胞（human umbilical vein endothelial cells,HUVECs）凋亡的影响及机制研究。方法: 运用平行平板流动腔建立LSS诱导的HUVECs凋亡模型。采用Hoechst33258荧光染色，流式细胞术检测CN对LSS诱导的HUVECs凋亡的影响。Western blot检测CN对LSS诱导的HUVECs中前蛋白转化酶枯草溶菌素9（proprotein convertase subtilisin/kexin type 9,PCSK9）蛋白表达的影响。结果:Hoechst33258荧光染色结果显示，与静态培养组比较，LSS组（3 dyne/cm2,12 h）内呈现凋亡状态的HUVECs明显增多。与LSS组比较，各CN+LSS组内呈现凋亡状态的HUVECs明显减少，并且呈现浓度依赖性。其中以20 μmol/L CN+LSS组凋亡细胞数最少；流式细胞术结果显示，与静态培养组比较，LSS组HUVECs凋亡率明显增加，差异有显著统计学意义（P<0.01）；与LSS组比较，各CN+LSS组HUVECs凋亡率明显减少，并且呈现浓度依赖性，其中以20 μmol/L CN+LSS组HUVECs凋亡率减少最为明显，差异具有显著统计学意义（P<0.01）。Western blot结果显示与静态培养组比较，LSS组内PCSK9蛋白表达显著升高，差异具有统计学意义(P<0.01)；与LSS组比较，CN+LSS组内PCSK9蛋白表达下降，并且呈现浓度依赖性，差异具有统计学意义（P<0.05）。其中以20 μmol/L CN+LSS组PCSK9蛋白表达下降最为明显(P<0.01)。结论: CN能够抑制LSS诱导的HUVECs凋亡，其作用机制与CN下调LSS处理的HUVECs中PCSK9蛋白表达相关。

关键词: 动脉粥样硬化, 烟酸姜黄素酯, 低剪切应力, 人脐静脉内皮细胞, 细胞凋亡, 前蛋白转化酶枯草溶菌素9

Abstract:

AIM: To investigate the effect of curcumin nicotinate（CN）on apoptosis of human umbilical vein endothelial cells (HUVECs) induced by low shear stress (LSS)and its mechanism. METHODS: A parallel plate flow chamber was used to establish a model of HUVECs apoptosis induced by low shear stress. Hoechst33258 fluorescence staining and flow cytometry were used to detect the effects of curcumin on the apoptosis of HUVECs. Western blot was used to detect the expression of proprotein convertase subtilisin/kexin type 9. RESULTS:Hoechst 33258 fluorescence staining showed that the number of HUVECs with apoptotic status in the LSS group was significantly higher than that in the static culture group. Compared with the LSS group, the HUVECs showing apoptotic status in the CN+LSS group were significantly reduced and showed a concentration-dependent manner. Among them, the least number of apoptotic cells was in 20 μmol/L CN+LSS group. Flow cytometry results showed that compared with the static culture group, the apoptosis rate of the LSS group was significantly increased, and the difference was statistically significant (P<0.01); compared with LSS group, the apoptosis rate of HUVECs in CN+LSS group was significantly reduced and showed a concentration-dependent manner. Among them, the decrease of apoptotic rate with 20 μmol/L CN+LSS group was the most obvious, the difference was statistically significant (P<0.01). Western blot results showed that compared with static culture group, PCSK9 protein expression was significantly increased in LSS group, and the difference was statistically significant (P<0.01). Compared with the LSS group, the expression of PCSK9 protein in the CN+LSS group was decreased and showed a concentrationdependent manner. The difference was statistically significant (P<0.05). Among them, the expression of PCSK9 protein was significantly decreased in 20 μmol/L CN+LSS group (P<0.01). CONCLUSION: CN inhibits the apoptosis of HUVECs induced by LSS, and its mechanism is related to down-regulation of the protein expression of PCSK9 in HUVECs under the action of LSS.

Key words: atherosclerosis, niacin curcumin esters, low shear stress, human umbilical vein endothelial cells, cell apoptosis, PCSK9

中图分类号:

R965.2

刘静，王媚媚，刘录山，贺卫和，庹勤慧，文红艳. 烟酸姜黄素酯对低剪切应力诱导人脐静脉内皮细胞凋亡的影响及相关机制[J]. 中国临床药理学与治疗学, 2018, 23(8): 855-861.

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烟酸姜黄素酯对低剪切应力诱导人脐静脉内皮细胞凋亡的影响及相关机制

Effects and mechanisms of nicotinic acid curcumin on apoptosis induced by low shear stress in human umbilical vein endothelial cells

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