Melatonin inhibits ox-LDL-induced proliferation of vascular smooth muscle cells

Yang TANG; Chuan LIU; Huali KANG; Jun JIN

doi:10.16016/j.1000-5404.202103139

您当前的位置：

首页 >

文章列表页 >

Melatonin inhibits ox-LDL-induced proliferation of vascular smooth muscle cells

更新时间：2026-01-09

- Melatonin inhibits ox-LDL-induced proliferation of vascular smooth muscle cells
- Journal of Third Military Medical University Vol. 43, Issue 17, Pages: 1619-1626(2021)
- 作者机构：
- 作者简介：
- 基金信息：
  
  Supported by the Chongqing Meritocratic Project for Leading Talents in Innovation and Entrepreneurship(2019)
- DOI：10.16016/j.1000-5404.202103139
  CLC：
- Received：17 March 2021，
  
  Published：15 September 2021
- 稿件说明：
移动端阅览
Yang TANG, Chuan LIU, Huali KANG, et al. Melatonin inhibits ox-LDL-induced proliferation of vascular smooth muscle cells[J]. Journal of Third Military Medical University, 2021, 43(17): 1619-1626.
DOI：

Yang TANG, Chuan LIU, Huali KANG, et al. Melatonin inhibits ox-LDL-induced proliferation of vascular smooth muscle cells[J]. Journal of Third Military Medical University, 2021, 43(17): 1619-1626. DOI： 10.16016/j.1000-5404.202103139.

摘要

目的

观察褪黑素（melatonin，Mel）对氧化型低密度脂蛋白（ox-LDL）诱导的血管平滑肌细胞（vascular smooth muscle cells，VSMCs）增殖功能的影响并探讨其可能的分子机制。

方法

① 选用不同浓度（0、10、20、50、100 μg/mL）的ox-LDL诱导VSMCs 48 h后检测细胞增殖功能，观察各浓度ox-LDL对VSMCs功能的影响。②选择最佳浓度（50 μg/mL）的ox-LDL诱导VSMCs 0、6、12、24、48 h后检测细胞增殖功能，观察各时间点ox-LDL对VSMCs功能的影响。③50 μg/mL的ox-LDL诱导VSMCs 24 h后，再用1、1×10

、1×10

nmol/L的Mel处理VSMCs 48 h，观察Mel作用不同时间对VSMCs增殖功能的影响。④使用ox-LDL联合Mel处理VSMCs。实验分4组：空白对照组（VSMCs正常培养不进行任何处理），Mel组（1×10

nmol/L Mel处理VSMCs 48 h），ox-LDL组（50 μg/mL ox-LDL诱导VSMCs 24 h），ox-LDL+ Mel组（使用50 μg/mL的ox-LDL诱导VSMCs 24 h后，接着使用1×10

nmol/L的Mel处理VSMCs 48 h）。采用CCK-8法检测细胞增殖，流式细胞术检测细胞周期，免疫荧光染色和Western blot检测增殖标记物（PCNA）、α平滑肌肌动蛋白（α-SMA）、细胞周期素A（Cyclin A）、细胞周期抑制蛋白p21以及细胞外调节蛋白激酶1/2（ERK1/2）信号通路相关蛋白表达。

结果

与空白对照组比较，50 μg/mL ox-LDL处理VSMCs 24 h能显著诱导细胞增殖，经ox-LDL（50 μg/mL）诱导24 h后，S期与G

期细胞显著增加，G

期细胞显著减少，促进细胞迁移，上调PCNA、α-SMA、Cyclin A、p-ERK1/2蛋白表达，并下调p-p21蛋白水平，差异均具有统计学意义（

0.01）。1、1×10

、1×10

nmol/L Mel均可不同程度抑制ox-LDL诱导的VSMCs增殖，与ox-LDL组比较，ox-LDL（50 μg/mL）处理VSMCs 24 h后接着使用Mel（1×10

nmol/L）处理48 h，可使G

期与S期细胞显著减少，G

期细胞显著增加，降低PCNA、α-SMA、Cyclin A、p-ERK1/2蛋白表达，上调p-p21蛋白表达，差异均具有统计学意义（

0.01）。

结论

Mel可显著抑制ox-LDL诱导的VSMCs的增殖，其作用机制可能与激活P21、阻断ERK1/2信号通路相关。

Abstract

Objective

To explore the effect of melatonin (Mel) on the proliferation of vascular smooth muscle cells (VSMCs) induced by oxidized low-density lipoprotein (ox-LDL) and investigate the potential molecular mechanism.

Methods

① ox-LDL at different concentrations (0

50 and 100 μg/mL) was used to treat VSMCs for 48 h

and cell proliferation was measured with CCK-8 assay. ② The effect of different time points (0

24 and 48 h) on the cell viability was also detect after the VSMCs was treated with the optional concentration of ox-LDL (50 μg/mL). ③ After the VSMCs were induced by 50 μg/mL ox-LDL for 24 h

the cells were treated with Mel at 1

1×10

and 1×10

nmol/L for 48 h to determine the effect of Mel on cell viability. ④ The VSMCs were divided into 4 groups

that is

blank control group

Mel group (1×10

nmol/L Mel for 48 h)

ox-LDL group (50 μg/mL ox-LDL for 24 h)

and ox-LDL+Mel group (50 μg/mL ox-LDL for 24 h followed by 1×10

nmol/L Mel for 48 h). CCK-8 assay and flow cytometry were employed to detect cell proliferation and cell cycle

respectively. Immunofluorescence staining and Western blotting were adopted to determine the expression of cell proliferation markers

proliferating cell nuclear antigen (PCNA) and α-smooth muscle actin (α-SMA)

cell cycle-related protein cyclin A (Cyclin A) and cyclin-dependent kinase inhibitor p21

as well as extracellular regulatory protein kinase 1/2 (ERK1/2).

Results

Compared with the blank control group

treatment of 50 μg/mL ox-LDL for 24 h in VSMCs significantly promoted cell proliferation

induced cell arrested at S and G

phase

decreased the proportion of cells at G

phase

improved cell migration

up-regulated the expression of PCNA

α-SMA

Cyclin A and p-ERK1/2

and down-regulated that of p-p21 protein (all

0.01). Mel of 1

1×10

and 1×10

nmol/L inhibited the prolife

rative ability of VSMCs induced by ox-LDL treatment. Compared with the ox-LDL group

the ox-LDL+Mel group had significantly reduced proportion of G

and S phase cells

enhanced proportion of G

phase cells

decreased protein levels of PCNA

α-SMA

Cyclin A and p-ERK1/2

and increased level of p-p21 (

0.01).

Conclusion

Mel significantly inhibits the proliferation in VSMCs induced by ox-LDL

which might be related to activation of P21 and block of ERK1/2 signaling pathway.

关键词

Keywords

references

PARMA L BAGANHA F QUAX P H A Plaque angiogenesis and intraplaque hemorrhage in atherosclerosis Eur J Pharmacol 2017 816 107 115 10.1016/j.ejphar.2017.04.028 PARMA L, BAGANHA F, QUAX P H A, et al . Plaque angiogenesis and intraplaque hemorrhage in atherosclerosis[J ] . Eur J Pharmacol, 2017, 816: 107-115. DOI: 10.1016/j.ejphar.2017.04.028.

JAIN T NIKOLOPOULOU E A XU Q B Hypoxia inducible factor as a therapeutic target for atherosclerosis Pharmacol Ther 2018 183 22 33 10.1016/j.pharmthera.2017.09.003 JAIN T, NIKOLOPOULOU E A, XU Q B, et al . Hypoxia inducible factor as a therapeutic target for atherosclerosis[J ] . Pharmacol Ther, 2018, 183: 22-33. DOI: 10.1016/j.pharmthera.2017.09.003.

LACOLLEY P REGNAULT V NICOLETTI A The vascular smooth muscle cell in arterial pathology: a cell that can take on multiple roles Cardiovasc Res 2012 95 2 194 204 10.1093/cvr/cvs135 LACOLLEY P, REGNAULT V, NICOLETTI A, et al . The vascular smooth muscle cell in arterial pathology: a cell that can take on multiple roles[J ] . Cardiovasc Res, 2012, 95(2): 194-204. DOI : 10.1093/cvr/cvs135.

袁帅廖思聪王大新血管平滑肌细胞与动脉粥样硬化关系的研究进展实用心脑肺血管病杂志 2018 26 2 6 8, 12 10.3969/j.issn.1008-5971.2018.02.002 袁帅, 廖思聪, 王大新. 血管平滑肌细胞与动脉粥样硬化关系的研究进展[J]. 实用心脑肺血管病杂志, 2018, 26(2): 6-8, 12. DOI: 10.3969/j.issn.1008-5971.2018.02.002.

张强徐立 Ox-LDL在动脉粥样硬化中的致病机制及检测方法研究进展国际检验医学杂志 2018 39 19 2432 2436 10.3969/j.issn.1673-4130.2018.19.025.ZHANGQ, 张强, 徐立. Ox-LDL在动脉粥样硬化中的致病机制及检测方法研究进展[J]. 国际检验医学杂志, 2018, 39(19): 2432-2436. DOI: 10.3969/j.issn.1673-4130.2018.19.025.

ZHAO T ZHANG H JIN C Melatonin mediates vasodilation through both direct and indirect activation of BKCa channels J Mol Endocrinol 2017 59 3 219 233 10.1530/JME-17-0028 ZHAO T, ZHANG H, JIN C, et al . Melatonin mediates vasodilation through both direct and indirect activation of BKCa channels[J ] . J Mol Endocrinol, 2017, 59(3): 219-233. DOI: 10.1530/JME-17-0028.

刘振华李步潆段维勋褪黑素抑制磷酸钙诱导的大鼠主动脉血管平滑肌细胞凋亡中国体外循环杂志 2016 14 2 112 115, 119 10.13498/j.cnki.chin.j.ecc.2016.02.13 刘振华, 李步潆, 段维勋, 等. 褪黑素抑制磷酸钙诱导的大鼠主动脉血管平滑肌细胞凋亡[J]. 中国体外循环杂志, 2016, 14(2): 112-115, 119. DOI: 10.13498/j.cnki.chin.j.ecc.2016.02.13.

ZHANG Y LIU N M WANG Y Endothelial cell calpain as a critical modulator of angiogenesis Biochim Biophys Acta Mol Basis Dis 2017 1863 6 1326 1335 10.1016/j.bbadis.2017.03.021 ZHANG Y, LIU N M, WANG Y, et al . Endotheli al cell calpain as a critical modulator of angiogenesis[J ] . Biochim Biophys Acta Mol Basis Dis, 2017, 1863(6): 1326-1335. DOI: 10.1016/j.bbadis.2017.03.021.

雷灵亮贝俊杰动脉粥样硬化斑块内血管生成的研究进展及治疗策略心血管病学进展 2019 40 3 478 482 10.16806/j.cnki.issn.1004-3934.2019.03.044 雷灵亮, 贝俊杰. 动脉粥样硬化斑块内血管生成的研究进展及治疗策略[J]. 心血管病学进展, 2019, 40(3): 478-482. DOI: 10.16806/j.cnki.issn.1004-3934.2019.03.044.

CORNELISSEN A GUO L SAKAMOTO A New insights into the role of iron in inflammation and atherosclerosis EBio Medicine 2019 47 598 606 10.1016/j.ebiom.2019.08.014 CORNELISSEN A, GUO L, SAKAMOTO A, et al . New insights into the role of iron in inflammation and atherosclerosis[J ] . EBio Medicine, 2019, 47: 598-606. DOI: 10.1016/j.ebiom.2019.08.014.

KWON H LEE J J LEE J H Cinnamon and its components suppress vascular smooth muscle cell proliferation by up-regulating cyclin-dependent kinase inhibitors Am J Chin Med 2015 43 4 621 636 10.1142/S0192415X1550038X KWON H, LEE J J, LEE J H, et al . Cinnamon and its components suppress vascular smooth muscle cell proliferation by up-regulating cyclin-dependent kinase inhibitors[J ] . Am J Chin Med, 2015, 43(4): 621-636. DOI: 10.1142/S0192415X1550038X.

OZER N K TAHA S AZZI A Homocysteine induces DNA synthesis and proliferation of vascular smooth muscle cells by interfering with MAPK kinase pathway Biofactors 2005 24 1/2/3/4 193 199 10.1002/biof.5520240123 OZER N K, TAHA S, AZZI A. Homocysteine induces DNA synthesis and proliferation of vascular smooth muscle cells by interfering with MAPK kinase pathway[J]. Biofactors, 2005, 24(1/2/3/4): 193-199. DOI: 10.1002/biof.5520240123.

WANG S C PCNA: a silent housekeeper or a potential therapeutic target? Trends Pharmacol Sci 2014 35 4 178 186 10.1016/j.tips.2014.02.004 WANG S C. PCNA: a silent housekeeper or a potential therapeutic target? [J]. Trends Pharmacol Sci, 2014, 35(4): 178-186. DOI: 10.1016/j.tips.2014.02.004.

SHINDE A V HUMERES C FRANGOGIANNIS N G The role of α-smooth muscle actin in fibroblast-mediated matrix contraction and remodeling Biochim Biophys Acta Mol Basis Dis 2017 1863 1 298 309 10.1016/j.bbadis.2016.11.006 SHINDE A V, HUMERES C, FRANGOGIANNIS N G. The role of α-smooth muscle actin in fibroblast-mediated matrix contraction and remodeling[J]. Biochim Biophys Acta Mol Basis Dis, 2017, 1863(1): 298-309. DOI: 10.1016/j.bbadis.2016.11.006.

ELKADY A I HUSSEIN R A ABU-ZINADAH O A Differential control of growth, apoptotic activity and gene expression in human colon cancer cells by extracts derived from medicinal herbs, Rhazya stricta and Zingiber officinale and their combination World J Gastroenterol 2014 20 41 15275 15288 10.3748/wjg.v20.i41.15275 ELKADY A I, HUSSEIN R A, ABU-ZINADAH O A. Differential control of growth, apoptotic activity and gene expression in human colon cancer cells by extracts derived from medicinal herbs, Rhazya stricta and Zingiber officinale and their combination[J]. World J Gastroenterol, 2014, 20(41): 15275-15288. DOI: 10.3748/wjg.v20.i41.15275.

徐丽金清东宫喜褪黑素的抗胃癌效应及其引起的Bcl-2、Bax、p21和p53的表达变化生理学报 2014 66 6 723 729 10.13294/j.aps.2014.0086 徐丽, 金清东, 宫喜, 等. 褪黑素的抗胃癌效应及其引起的Bcl-2、Bax、p21和p53的表达变化[J]. 生理学报, 2014, 66(6): 723-729. DOI: 10.13294/j.aps.2014.0086.

YU S CHEN Y CHEN S Klotho inhibits proliferation and migration of angiotensin Ⅱ-induced vascular smooth muscle cells (VSMCs) by modulating NF-κB p65, Akt, and extracellular signal regulated kinase (ERK) signaling activities Med Sci Monit 2018 24 4851 4860 10.12659/msm.908038 YU S, CHEN Y, CHEN S, et al . Klotho inhibits proliferation and migration of angiotensin Ⅱ-induced vascular smooth muscle cells (VSMCs) by modulatin g NF-κB p65, Akt, and extracellular signal regulated kinase (ERK) signaling activities[J ] . Med Sci Monit, 2018, 24: 4851-4860. DOI: 10.12659/msm.908038.

LI T WANG F DANG Y P21 and P27 promote tumorigenesis and progression via cell cycle acceleration in seminal vesicles of TRAMP mice Int J Biol Sci 2019 15 10 2198 2210 10.7150/ijbs.35092 LI T, WANG F, DANG Y, et al . P21 and P27 promote tumorigenesis and progression via cell cycle acceleration in seminal vesicles of TRAMP mice[J ] . Int J Biol Sci, 2019, 15(10): 2198-2210. DOI:10.7150/ijbs.35092.

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Effects of different levels of overworking on rat arterial smooth muscle cells

Role of estrogen receptor α in process of insulin-induced atherosclerosis

Effect of exogenous apelin-13 on formation of atherosclerosis in rats

D₄ dopamine receptor suppresses AngⅡ-mediated proliferation of vascular smooth muscle cells in vitro

Expression of miR-616 in osteosarcoma and its role in proliferation, apoptosis, migration and invasion of tumor cells

Related Author

Lu GAN

Suheng CHEN

Miao ZHUANG

Xiaoxiao ZHANG

Hong GUO

Rongrong HUANG

Yulan LI

仲伟天

Related Institution

The First Clinical Medical College, Lanzhou University

Department of Anesthesiology, the First Hospital of Lanzhou University

Department of Pathology， Second Affiliated Hospital， Army Medical University （Third Military Medical University）

Guangxi Key Laboratory of Characteristic Biomedicine， School of Medicine， Guangxi University

Scientific Research Experiment Center， Chengdu Medical College

⁰