Overexpression of SCAP inhibits autophagy and promotes lipid accumulation in vascular smooth muscle cells

Yuhan RAO; Danyang LI; Xiaoli HOU; Yincheng YAO; Yu SU; Yaxi CHEN; Xiongzhong RUAN

doi:10.16016/j.2097-0927.202110042

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Overexpression of SCAP inhibits autophagy and promotes lipid accumulation in vascular smooth muscle cells

Basic Medicine | 更新时间：2026-01-09

- Overexpression of SCAP inhibits autophagy and promotes lipid accumulation in vascular smooth muscle cells
- Journal of Army Medical University Vol. 44, Issue 9, Pages: 891-897(2022)
- 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：
- 基金信息：
  
  the National Natural Science Foundation for Young Scholars of China(81900406)
- DOI：10.16016/j.2097-0927.202110042
  CLC： R341;R363.21;R543.5
- Received：11 October 2021，
  
  Revised：2022-01-05，
  
  Published：15 May 2022
- 稿件说明：
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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：

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

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

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

与LV-Control组相比，LV-SCAP组nSREBP2表达明显上调(

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(

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(

0.05)。Rapa处理结果显示LC3b表达增加，P62水平降低(

0.05)，油红O染色脂滴沉积减少(

0.05)。

结论

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

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

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

Compared with LV-Control group

the protein expression of nSREBP2 was increased after the infection of lentiviral vector (

0.05). LV-SCAP group also presented accelerated accumulation of lipid droplets and increased contents of TC and TG (

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 (

0.05). Rapa treatment resulted in enhanced expression of LC3b and reduced level of P62 (

0.05)

and decreased accumulation of lipid droplets (

0.05).

Conclusion

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

LIBBY P, BU RING J E, BADIMON L, et al . Atherosclerosis[J ] . Nat Rev Dis Primers, 2019, 5(1): 56. DOI: 10.1038/s41572-019-0106-z.

BENTZON J F, OTSUKA F, VIRMANI R, et al . Mechanisms of plaque formation and rupture[J ] . Circ Res, 2014, 114(12): 1852-1866. DOI: 10.1161/CIRCRESAHA.114.302721.

GOLDSTEIN J L, BROWN M S. A century of cholesterol and coronaries: from plaques to genes to statins[J]. Cell, 2015, 161(1): 161-172. DOI: 10.1016/j.cell.2015.01.036.

JOHNSTON J M, ANGYAL A, BAUER R C, et al . Myeloid Tribbles 1 induces early atherosclerosis via enhanced foam cell expansion[J ] . Sci Adv, 2019, 5(10): eaax9183. DOI: 10.1126/sciadv.aax9183.

MENG X D, YAO H H, WANG L M, et al . Knockdown of GAS5 inhibits atherosclerosis progression via reducing EZH2-mediated ABCA1 transcription in ApoE -/- mice[J ] . Mol Ther Nucleic Acids, 2020, 19: 84-96. DOI: 10.1016/j.omtn.2019.10.034..

GROOTAERT M O J, MOULIS M, ROTH L, et al . Vascular smooth muscle cell death, autophagy and senescence in atherosclerosis[J ] . Cardiovasc Res, 2018, 114(4): 622-634. DOI: 10.1093/cvr/cv y007.

GUERRINI V, GENNARO M L. Foam cells: one size doesn't fit all[J]. Trends Immunol, 2019, 40(12): 1163-1179. DOI: 10.1016/j.it.2019.10.002.

LIU Z Y, ZHU H P, DAI X Y, et al . Macrophage liver kinase B1 inhibits foam cell formation and atherosclerosis[J ] . Circ Res, 2017, 121(9): 1047-1057. DOI: 10.1161/CIRCRESAHA.117.311546.

CANFRÁN-DUQUE A, ROTLLAN N, ZHANG X B, et al . Macrophage deficiency of miR-21 promotes apoptosis, plaque necrosis, and vascular inflammation during atherogenesis[J ] . EMBO Mol Med, 2017, 9(9): 1244-1262. DOI: 10.15252/emmm.201607492.

ALLAHVERDIAN S, CHEHROUDI A C, MCMANUS B M, et al . Contribution of intimal smooth muscle cells to cholesterol accumulation and macrophage-like cells in human atherosclerosis[J ] . Circulation, 2014, 129(15): 1551-1559. DOI: 10.1161/CIRCULATIONAHA.113.005015.

BROWN M S, RADHAKRISHNAN A, GOLDSTEIN J L. Retrospective on cholesterol homeostasis: the central role of scap[J]. Annu Rev Biochem, 2018, 87: 783-807. DOI: 10.1146/annurev-biochem-062917-011852.

YABE D, XIA Z P, ADAMS C M, et al . Three mutations in sterol-sensing domain of SCAP block interaction with insig and render SREBP cleavage insensitive to sterols[J ] . Proc Natl Acad Sci USA, 2002, 99(26): 16672-16677. DOI: 10.1073/pnas.262669399.

LIU X P, TANG Y, CUI Y C, et al . Autophagy is associated with cell fate in the process of macrophage-derived foam cells formation and progress[J ] .J Biomed Sci, 2016, 23(1): 57. DOI: 10.1186/s12929-016-0274-z.

HE J P, ZHANG G Y, PANG Q, et al . SIRT6 reduces macrophage foam cell formation by inducing autophagy and cholesterol efflux under ox-LDL condition[J ] . FEBS J, 2017, 284(9): 1324-1337. DOI: 10.1111/febs.14055.

GÓMEZ-SÁNCHEZ R, YAKHINE-DIOP S M, RODRÍGUEZ-ARRIBAS M, et al . mRNA and protein dataset of autophagy markers (LC3 and p62) in several cell lines[J ] . Data Brief, 2016, 7: 641-647. DOI: 10.1016/j.dib.2016.02.085.

CHEN M L, YI L, JIN X, et al . Resveratrol attenuates vascular endothelial inflammation by inducing autophagy through the cAMP signaling pathway[J ] . Autophagy, 2013, 9(12): 2033-2045. DOI: 10.4161/auto.26336.

LI G, SCULL C, OZCAN L, et al . NADPH oxidase links endoplasmic Reticulum stress, oxidative stress, and PKR activation to induce apoptosis[J ] . J Cell Biol, 2010, 191(6): 1113-1125. DOI: 10.1083/jcb.201006121.

HAN X B, LI H X, JIANG Y Q, et al . Upconversion nanoparticle-mediated photodynamic therapy induces autophagy and cholesterol efflux of macrophage-derived foam cells via ROS generation[J ] . Cell Death Dis, 2017, 8(6): e2864. DOI: 10.1038/cddis.2017.242.

TAMARGO-GÓMEZ I, MARIÑO G. AMPK: regulation of metabolic dynamics in the context of autophagy[J]. Int J Mol Sci, 2018, 19(12): E3812. DOI: 10.3390/ijms19123812.

KIM J, KUNDU M, VIOLLET B, et al . AMPK and mTOR regulate autophagy through direct phosphorylation of Ulk1[J ] . Nat Cell Biol, 2011, 13(2): 132-141. DOI: 10.1038/ncb2152.

CHEN H, JI Y S, YAN X, et al . Berberine attenuates apoptosis in rat retinal Müller cells stimulated with high glucose via enhancing autophagy and the AMPK/mTOR signaling[J ] . Biomed Pharmacother, 2018, 108: 1201-1207. DOI: 10.1016/j.biopha.2018.09.140.

KIM Y C, GUAN K L. mTOR: a pharmacologic target for autophagy regulation[J]. J Clin Invest, 2015, 125(1): 25-32. DOI: 10.1172/JCI73939.

LI D Y, CHEN A M, LAN T, et al . SCAP knockdown in vascular smooth muscle cells alleviates atherosclerosis plaque formation via up-regulating autophagy in ApoE -/- mice[J ] . FASEB J, 2019, 33(3): 3437-3450. DOI:10.1096/fj.201800975RRR..

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