[1]张英,陈林,陈杨,等.NOX/ROS在5-HT介导大鼠肺动脉平滑肌细胞增殖中的作用[J].陆军军医大学学报(原第三军医大学学报),2020,42(24):2382-2386.
 ZHANG Ying,CHEN Lin,CHEN Yang,et al.Role of NOX/ROS in 5-HT-mediated proliferation of rat pulmonary artery smooth muscle cells[J].J Amry Med Univ (J Third Mil Med Univ),2020,42(24):2382-2386.
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
42卷
期数:
2020年第24期
页码:
2382-2386
栏目:
基础医学
出版日期:
2020-12-30

文章信息/Info

Title:
Role of NOX/ROS in 5-HT-mediated proliferation of rat pulmonary artery smooth muscle cells
作者:
张英陈林陈杨易斌陈杰
陆军军医大学(第三军医大学)第一附属医院麻醉科;重庆市巴南区人民医院手术麻醉科
Author(s):
ZHANG Ying CHEN Lin CHEN Yang YI Bin CHEN Jie
Department of Anesthesiology, First Affiliated Hospital, Army Medical University (Third Military Medical University), Chongqing, 400038; 2Department of Surgical Anesthesiology,Banan District People’s Hospital, Chongqing, 401320, China
 
关键词:
 
Keywords:
5-HT NADPH oxidase reactive oxygen species ERK1/2 pulmonary artery smooth muscle cells
分类号:
R322.121;R329.28;R363.21
文献标志码:
A
摘要:

目的研究5-羟色胺(5-HT)介导的肺动脉平滑肌细胞增殖是否与细胞内活性氧(reactive oxygen species, ROS)升高有关, 并明确NADPH氧化酶(NADPH oxidase, NOX)在ROS产生及可能由此导致的肺动脉平滑肌细胞增殖中的作用。方法组织块法建立人肺动脉平滑肌细胞(pulmonary artery smooth muscle cells, PASMCs)细胞系。分别设置正常对照组,不加任何药物; 5-HT组,给予5-HT(1 μmol/L)刺激细胞;各干预组分别给予ROS清除剂Tempol(100 μmol/L)、NOX抑制剂DPI(20 μmol/L)、ERK抑制剂PD98059(30 μmol/L) 预处理,再加入5-HT刺激不同时间。处理完成后测定各组NOX活性,采用DCF-DA荧光探针检测细胞内ROS的生成,3H-TdR掺入法检测PASMCs的增殖,免疫印迹杂交法检测ERK1/2磷酸化水平。结果研究发现5-HT刺激可明显增强NOX活性(P<0.05),引起ROS的增加(P<0.05),DNA合成增加(P<0.05),ERK1/2磷酸化水平增加(P<0.05)。NOX抑制剂(DPI)、ROS清除剂(Tempol)或ERK抑制剂PD98059均能显著地逆转5-HT刺激引起的ERK1/2磷酸化及DNA合成水平的增加(P<0.05),NOX抑制剂(DPI)能显著地逆转5-HT刺激引起的ROS增加(P<0.05)。结论NOX/ROS可能是5-HT介导的PASMCs增殖信号转导重的重要蛋白

Abstract:
ObjectiveTo observe whether serotonin (5-HT)-mediated proliferation of pulmonary artery smooth muscle cells (PASMCs) is related to the increase of intracellular reactive oxygen species (ROS), and identify the role of NADPH oxidase (NOX) in the production of ROS and 5-HT-mediated proliferation. MethodsTissue block culture method was used to establish a cell line of human PASMCs. Then the cells were divided into 6 groups, normal control (no treatment), 5-HT treatment (1 μmol/L 5-HT), and 3 intervention groups [with pretreatment of ROS scavenger Tempol (100 μmol/L), NOX inhibitor DPI (20 μmol/L) and ERK inhibitor PD98059 (30 μmol/L) for 30 min, respectively and then 5-HT stimulation for different time periods]. Then NOX activity was analyzed by using spectrophotometer to calculate the amount of NADPH consumed. ROS production was measured by DCF-DA assay. The expression of phosphorylated-ERK1/2 (p-ERK1/2) was detected by Western blotting. The changes of PASMC proliferation were determined 3H-TdR incorporation assay. Results5-HT stimulation for 30 min obviously enhanced the NOX activity, ROS production, DNA synthesis, and expression of p-ERK1/2 (all P<0.05). While the pretreatment of DPI, Tempol or PD98059 could significantly reverse the ERK1/2 phosphorylation and DNA synthesis induced by 5-HT stimulation (both P<0.05). What’s more, DPI also notably reduced the generation of ROS (P<0.05). ConclusionNOX/ROS may be the key proteins in the signaling pathways of 5-HT mediated proliferation of PASMCs.
 

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更新日期/Last Update: 2020-12-22