褪黑素抑制氧化型低密度脂蛋白诱导血管平滑肌细胞增殖

唐杨; 刘川; 康华利; 晋军

doi:10.16016/j.1000-5404.202103139

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褪黑素抑制氧化型低密度脂蛋白诱导血管平滑肌细胞增殖

基础医学 | 更新时间：2026-01-09

- 褪黑素抑制氧化型低密度脂蛋白诱导血管平滑肌细胞增殖
- Melatonin inhibits ox-LDL-induced proliferation of vascular smooth muscle cells
- 第三军医大学学报 2021年43卷第17期页码：1619-1626
- 作者机构：
- 作者简介：
- 基金信息：
  
  重庆英才·创新创业领军人才项目(2019)
- DOI：10.16016/j.1000-5404.202103139
  中图分类号：
- 收稿：2021-03-17，
  
  纸质出版：2021-09-15
- 稿件说明：
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唐杨, 刘川, 康华利, 等. 褪黑素抑制氧化型低密度脂蛋白诱导血管平滑肌细胞增殖[J]. 第三军医大学学报, 2021,43(17):1619-1626.

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.
唐杨, 刘川, 康华利, 等. 褪黑素抑制氧化型低密度脂蛋白诱导血管平滑肌细胞增殖[J]. 第三军医大学学报, 2021,43(17):1619-1626. DOI： 10.16016/j.1000-5404.202103139.

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

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