NOX/ROS在5-HT介导大鼠肺动脉平滑肌细胞增殖中的作用

张英; 陈林; 陈杨; 易斌; 陈杰

doi:10.16016/j.1000-5404.202006132

您当前的位置：

首页 >

文章列表页 >

NOX/ROS在5-HT介导大鼠肺动脉平滑肌细胞增殖中的作用

基础医学 | 更新时间：2026-01-09

- NOX/ROS在5-HT介导大鼠肺动脉平滑肌细胞增殖中的作用
- Role of NOX/ROS in 5-HT-mediated proliferation of rat pulmonary artery smooth muscle cells
- 第三军医大学学报 2020年42卷第24期页码：2382-2386
- 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：
  
  [ 易斌，E-mail:yibin1974@163.com YI Bin, E-mail:yibin1974@163.com
- 基金信息：
- DOI：10.16016/j.1000-5404.202006132
  中图分类号： R322.121;R329.28;R363.21
- 收稿：2020-06-14，
  
  修回：2020-10-12，
  
  纸质出版：2020-12-30
- 稿件说明：
移动端阅览
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]. 第三军医大学学报, 2020,42(24):2382-2386.

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.
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]. 第三军医大学学报, 2020,42(24):2382-2386. DOI： 10.16016/j.1000-5404.202006132.

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

HAN J L, TIAN H Y, LIU Y, et al . 5-HT promotes pulmonary arterial smooth muscle cell proliferation through the TRPC channel[J ] . Cell Mol Biol (Noisy-le-grand), 2018, 64(13): 89. DOI: 10.14715/cmb/2018.64.13.17.

HOOD K Y, MAIR K M, HARVEY A P, et al . Serotonin signaling through the 5-HT 1B receptor and NADPH oxidase 1 in pulmonary arterial hypertension[J ] . Arterioscler Thromb Vasc Biol, 2017, 37(7): 1361-1370. DOI: 10.1161/atvbaha.116.308929.

LI T, LUO X J, WANG E L, et al . Magnesium lithospermate B prevents phenotypic transformation of pulmonary arteries in rats with hypoxic pulmonary hypertension through suppression of NADPH oxidase[J ] . Eur J Pharmacol, 2019, 847: 32-41. DOI: 10.1016/j.ejphar.2019.01.020.

GOLOVINA V A, BLAUSTEIN M P. Preparation of primary cultured mesenteric artery smooth muscle cells for fluorescent imaging and physiological studies[J]. Nat Protoc, 2006, 1(6): 2681-2687. DOI: 10.1038/nprot.2006.425.

HERRERA B, MURILLO M M, ÁLVAREZ-BARRIENTOS A, et al . Source of early reactive oxygen species in the apoptosis induced by transforming growth factor-β in fetal rat hepatocytes[J ] . Free Radic Biol Med, 2004, 36(1): 16-26. DOI: 10.1016/j.freeradbiomed.2003.09.020.

BOPP C, AUGER C, DIEMUNSCH P, et al . The effect of urapidil, an alpha-1 adrenoceptor antagonist and a 5-HT1A agonist, on the vascular tone of the porcine coronary and pulmonary arteries, the rat aorta and the human pulmonary artery[J ] . Eur J Pharmacol, 2016, 779: 53-58. DOI: 10.1016/j.ejphar.2016.03.012.

ZIMMER A, TEIXEIRA R B, BONETTO J H P, et al . Role of inflammation, oxidative stress, and autonomic nervous system activation during the development of right and left cardiac remodeling in experimental pulmonary arterial hypertension[J ] . Mol Cell Biochem, 2020, 464(1/2): 93-109. DOI: 10.1007/s11010-019-03652-2.

YOU B Y, LIU Y B, CHEN J, et al . Vascular peroxidase 1 mediates hypoxia-induced pulmonary artery smooth muscle cell proliferation, apoptosis resistance and migration[J ] . Cardiovasc Res, 2018, 114(1): 188-199. DOI: 10.1093/cvr/cvx234.

PETRY A, GÖRLACH A. Regulation of NADPH oxidases by G protein-coupled receptors[J]. Antioxid Redox Signal, 2019, 30(1): 74-94. DOI: 10.1089/ars.2018.7525.

GUIGNABERT C, IZIKKI M, TU L I, et al . Transgenic mice overexpressing the 5-hydroxytryptamine transporter gene in smooth muscle develop pulmonary hypertension[J ] . Circ Res, 2006, 98(10): 1323-1330. DOI: 10.1161/01.res.0000222546.45372.a0.

IZIKKI M, HANOUN N, MARCOS E, et al . Tryptophan hydroxylase 1 knockout and tryptophan hydroxylase 2 polymorphism: effects onhypoxic pulmonary hypertension in mice[J ] . Am J Physiol Lung Cell Mol Physiol, 2007, 293(4): L1045-L1052. DOI: 10.1152/ajplung.00082.2007.

DELANEY C, SHERLOCK L, FISHER S, et al . Serotonin 2A receptor inhibition protects against the development of pulmonary hypertension and pulmonary vascular remodeling in neonatal mice[J ] . Am J Physiol Lung Cell Mol Physiol, 2018, 314(5): L871-L881. DOI: 10.1152/ajplung.00215.2017.

JIAO Y R, WANG W, LEI P C, et al . 5-HTT, BMPR2, EDN1, ENG, KCNA5 gene polymorphisms and susceptibility to pulmonary arterial hypertension: a meta-analysis[J ] . Gene, 2019, 680: 34-42. DOI: 10.1016/j.gene.2018.09.020.

MAGNANI F, MATTEVI A. Structure and mechanisms of ROS generation by NADPH oxidases[J]. Curr Opin Struct Biol, 2019, 59: 91-97. DOI: 10.1016/j.sbi.2019.03.001.

ZHANG L X, MA C, GU R, et al . Paeonol regulates hypoxia-induced proliferation of pulmonary artery smooth muscle cells via EKR 1/2 signalling[J ] . Eur J Pharmacol, 2018, 834: 257-265. DOI: 10.1016/j.ejphar.2018.07.017.

YANG X D, LONG L, SOUTHWOOD M, et al . Dysfunctional smad signaling contributes to abnormal smooth muscle cell proliferation in familial pulmonary arterial hypertension[J ] . Circ Res, 2005, 96(10): 1053-1063. DOI: 10.1161/01.res.0000166926.54293.68.

KOVACS L, HAN W H, RAFIKOV R, et al . Activation of calpain-2 by mediators in pulmonary vascular remodeling of pulmonary arterial hypertension[J ] . Am J Respir Cell Mol Biol, 2016, 54(3): 384-393. DOI: 10.1165/rcmb.2015-0151oc.

浏览量

下载量

CSCD

文章被引用时，请邮件提醒。

提交

工具集

关联资源

帕罗西汀与克龄蒙联合治疗围绝经期抑郁症5-羟色胺、雌二醇变化及疗效观察

牛磺酸对肺动脉内皮和肺动脉平滑肌细胞增殖的双向调节作用

不同方法培养大鼠肺动脉平滑肌细胞的差异

低氧对大鼠肺动脉平滑肌细胞中Gax基因表达及细胞增殖的影响

脊髓压迫伤后髓内5-HT和5-HIAA的变化