Role of NOX/ROS in 5-HT-mediated proliferation of rat pulmonary artery smooth muscle cells

Ying ZHANG; Lin CHEN; Yang CHEN; Bin YI; Jie CHEN

doi:10.16016/j.1000-5404.202006132

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Role of NOX/ROS in 5-HT-mediated proliferation of rat pulmonary artery smooth muscle cells

Basic Medicine | 更新时间：2026-01-09

- Role of NOX/ROS in 5-HT-mediated proliferation of rat pulmonary artery smooth muscle cells
- Journal of Third Military Medical University Vol. 42, Issue 24, Pages: 2382-2386(2020)
- 作者机构：
  
  1.400038 重庆，陆军军医大学(第三军医大学)第一附属医院麻醉科
  2.401320 重庆，重庆市巴南区人民医院手术麻醉科
- 作者简介：
- 基金信息：
- DOI：10.16016/j.1000-5404.202006132
  CLC： R322.121;R329.28;R363.21
- Received：14 June 2020，
  
  Revised：2020-10-12，
  
  Published：30 December 2020
- 稿件说明：
移动端阅览
Ying ZHANG, Lin CHEN, Yang CHEN, et al. Role of NOX/ROS in 5-HT-mediated proliferation of rat pulmonary artery smooth muscle cells[J]. Journal of Third Military Medical University, 2020, 42(24): 2382-2386.
DOI：

Ying ZHANG, Lin CHEN, Yang CHEN, et al. Role of NOX/ROS in 5-HT-mediated proliferation of rat pulmonary artery smooth muscle cells[J]. Journal of Third Military Medical University, 2020, 42(24): 2382-2386. DOI： 10.16016/j.1000-5404.202006132.

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(

0.05)，引起ROS的增加(

0.05)，DNA合成增加(

0.05)，ERK1/2磷酸化水平增加(

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(

0.05)，NOX抑制剂(DPI)能显著地逆转5-

HT刺激引起的ROS增加(

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

Objective

To 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.

Methods

Tissue 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. 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.

Results

5-HT stimulation for 30 min obviously enhanced the NOX activity

ROS production

DNA synthesis

and expression of p-ERK1/2 (all

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

0.05). What's more

DPI also notably reduced the generation of ROS (

0.05).

Conclusion

NOX/ROS may be the key proteins in the signaling pathways of 5-HT mediated proliferation of PASMCs.

关键词

Keywords

references

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