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血管平滑肌SCAP过表达抑制自噬促进脂质积聚的机制研究
Overexpression of SCAP inhibits autophagy and promotes lipid accumulation in vascular smooth muscle cells
- 陆军军医大学学报 2022年44卷第9期 页码：891-897
- 作者机构：
400016 重庆，重庆医科大学脂糖代谢性疾病重庆市重点实验室，脂质研究中心
- 作者简介：
[李丹阳，E-mail: lidycq@cqmu.edu.cn LI Danyang, E-mail: lidycq@cqmu.edu.cn
- 基金信息：国家自然科学基金青年基金项目(81900406)
DOI：10.16016/j.2097-0927.202110042
中图分类号： R341;R363.21;R543.5收稿：2021-10-11，
修回：2022-01-05，
纸质出版：2022-05-15
- 稿件说明：
移动端阅览
饶雨涵, 李丹阳, 侯晓俐, 等. 血管平滑肌SCAP过表达抑制自噬促进脂质积聚的机制研究[J]. 陆军军医大学学报, 2022,44(9):891-897.
Yuhan RAO, Danyang LI, Xiaoli HOU, et al. Overexpression of SCAP inhibits autophagy and promotes lipid accumulation in vascular smooth muscle cells[J]. Journal of Army Medical University, 2022, 44(9): 891-897.
饶雨涵, 李丹阳, 侯晓俐, 等. 血管平滑肌SCAP过表达抑制自噬促进脂质积聚的机制研究[J]. 陆军军医大学学报, 2022,44(9):891-897. DOI： 10.16016/j.2097-0927.202110042.
Yuhan RAO, Danyang LI, Xiaoli HOU, et al. Overexpression of SCAP inhibits autophagy and promotes lipid accumulation in vascular smooth muscle cells[J]. Journal of Army Medical University, 2022, 44(9): 891-897. DOI： 10.16016/j.2097-0927.202110042.
摘要
目的
2
探讨固醇调节元件结合蛋白裂解活化蛋白(sterol regulatory element binding proteins cleavage activating protein，SCAP)对血管平滑肌细胞(vascular smooth muscle cells，VSMCs)泡沫化的影响及其分子机制。
方法
2
构建LV-SCAP与LV-Control血管平滑肌细胞系，免疫印迹法检测SCAP及其下游相关蛋白nSREBP2表达水平；油红O染色观察负荷LDL胆固醇后脂滴积聚，酶标仪检测细胞总胆固醇(total cholesterol，TC)、甘油三酯(triglycerides，TG)水平；单丹磺酰尸胺(monodansyl cadaverine，MDC)染色检测自噬小泡聚集；Western blot检测自噬相关蛋白LC3b、P62与AMPK-mTOR通路表达；mTOR抑制剂雷帕霉素处理LV-SCAP过表达细胞株，Western blot与油红O染色验证处理后效果。
结果
2
与LV-Control组相比，LV-SCAP组nSREBP2表达明显上调(
P
<
0.05)，经LDL胆固醇负荷后产生的脂滴增多，TC、TG水平明显增高(
P
<
0.05)；LV-SCAP组细胞内自噬小泡聚集减少，自噬相关蛋白LC3b下调，P62上调，p-AMPK/AMPK水平降低，p-mTOR/mTOR水平升高(
P
<
0.05)。Rapa处理结果显示LC3b表达增加，P62水平降低(
P
<
0.05)，油红O染色脂滴沉积减少(
P
<
0.05)。
结论
2
SCAP可能通过影响AMPK-mTOR信号通路抑制血管平滑肌细胞的自噬过程，促进其泡沫化，进而参与血管动脉粥样硬化的发生发展。
Abstract
Objective
2
To determine the effect of sterol regulatory element binding proteins cleavage activating protein (SCAP) on foam cell formation in vascular smooth muscle cells (VSMCs).
Methods
2
LV-SCAP were constructed with LV-SCAP
and LV-Control group served as control. The expression of SCAP and its downstream protein nSREBP2 was detected with Western blotting. Oil red O staining was used to observe lipid accumulation after LDL treatment
and the contents of total cholesterol (TC) and triglycerides (TG) were measured by Microplate Reader. The autophagic vacuoles were detected with monodansyl cadaverine (MDC) staining. The protein levels of autophagy-related proteins LC3b
P62
and AMPK-mTOR pathway were detected by Western blotting. After rapamycin (Rapa)
mTOR inhibitor
was used to treat LV-SCAP
Western blotting and oil red O staining were performed again to observe the changes in the autophagy and accumulation of lipid droplets.
Results
2
Compared with LV-Control group
the protein expression of nSREBP2 was increased after the infection of lentiviral vector (
P
<
0.05). LV-SCAP group also presented accelerated accumulation of lipid droplets and increased contents of TC and TG (
P
<
0.05)
decreased formation of autophagic vacuoles
down-regulation of LC3b an
d up-regulation of P62
decreased level of p-AMPK/AMPK
and increased level of p-mTOR/mTOR (
P
<
0.05). Rapa treatment resulted in enhanced expression of LC3b and reduced level of P62 (
P
<
0.05)
and decreased accumulation of lipid droplets (
P
<
0.05).
Conclusion
2
SCAP inhibits VSMCs autophagy through AMPK-mTOR signaling pathway
and promotes the foam cell formation to participate in the development of atherosclerosis.
关键词
Keywords
references
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