改构黄藤素对钙化细胞的作用机制研究
The underlying mechanism of modified palmatine for calcified cells
作者：乔洁(四川大学生命科学学院)；李怡(电子科技大学附属医院.四川省人民医院，四川省人民医院肾内科暨肾脏病研究所)；杨黎(四川大学华西医院协同创新中心)；齐庆荣(四川大学华西药学院药化系)；王莉(电子科技大学附属医院.四川省人民医院，四川省人民医院肾内科暨肾脏病研究所)；林宏辉(四川大学生命科学学院)
Author：QIAO Jie(College of Life Sciences,Sichuan University)；LI Yi(Renal Division and Institute of Nephrology, Sichuan Academy of Medical Science and Sichuan Provincial People's Hospital, School of Medicine, University of Electronic Science and Technology)；YANG Li(Cancer Center, West China Hospital, Sichuan University and Collaborative Innovation Center)；QI Qing-Rong(Department of Medici nal Chemistry, West China School of Pharmacy, Sichuan University)；WANG Li(Renal Division and Institute of Nephrology, Sichuan Academy of Medical Science and Sichuan Provincial People's Hospital, School of Medicine, University of Electronic Science and Technology)；LIN Hong-Hui(College of Life Sciences， Sichuan University)
收稿日期：2017-10-25 年卷（期）页码：2018,55(6):1319-1325
期刊名称：四川大学学报: 自然科学版
Journal Name：Journal of Sichuan University (Natural Science Edition)
关键字：血管钙化； 黄藤素； 氧化应激； 炎症； 核因子NF-E2相关因子
Key words：Vascular calcification; Palmatine; Oxidative stress; Inflammation; Nuclear transcription factor nuclear factor-erythroid 2-related factor 2
基金项目：国家自然科学基金（81770742)
中文摘要
通过建立β 半胱氨酸磷酸盐诱导大鼠血管平滑肌细胞（RASMCs）钙化的模型，研究改构黄藤素在血管钙化中的作用.通过 Von Kossa 染色技术检测发现改构黄藤素可以显著降低钙化细胞内的钙盐沉积；利用实时定量 PCR（Real time Quantitative PCR）技术检测发现改构黄藤素可以将钙化细胞内的 Nrf-2 mRNA 水平由原来的 0.09±0.01 显著升高到 2.54±0.35；采用 Western Blot 技术检测发现改构黄藤素可以显著增强钙化细胞内的蛋白表达量；试剂盒检测活性氧（ROS）的含量发现改构黄藤素可以将钙化细胞内的荧光强度由原来的1.04±0.03 显著降低到 0.82±0.01；试剂盒检测一氧化氮（NO）的含量，发现改构黄藤素可以将钙化细胞内的 NO 含量由原来的 5.73 ± 0.05 μmol/L 显著降低到3.73 ± 0.15 μmol/L.
英文摘要
To explore the role of palmatine in vascular calcification，we employed a calcific cell model using β-glyceophosphate to stimulate the rat aortic smooth muscle cells (RASMCs). We performed a Von Kossa staining assay to detect the effect of palmatine on calcific RASMCs upon calcium deposition, we found that the modified palmatine could significantly inhibit intracellular calcium deposition of RASMCs after calcification. The mRNA level of the nuclear transcription factor nuclear factor-erythroid 2-related factor 2 (Nrf-2) was determined by Real-time Quantitative PCR, we found that the modified palmatine could significantly increase the mRNA level of Nrf-2 from (0.09±0.01) fold to (2.54±0.35) fold in RASMCs following calcification. Further，we measured the protein level of Nrf-2 by Western Blot, we found that the modified palmatine could significantly activate the expression of Nrf-2 in RASMCs following calcification. We used a Reactive Oxygen Species (ROS) kit to detect the fluorescence intensity of ROS, we observed that the modified palmatine could respectively decrease the fluorescence intensity indicating ROS from (1.04±0.03) to (0.82±0.01) in RASMCs after calcification. We used a Nitric Oxide (NO) kit to measure the production of NO, we observed that the modified palmatine could decrease the production of NO from (5.73±0.05) μmol/L to (3.73±0.15) μmol/L in RASMCs after calcification.
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