Quercetin protects against atherosclerosis by regulating the expression of PCSK9, CD36, PPARγ, LXRα and ABCA1

Jia,Qingling; Cao,Hui; Shen,Dingzhu; Li,Shanshan; Yan,Li; Chen,Chuan; Xing,Sanli; Dou,Fangfang

doi:10.3892/ijmm.2019.4263

Quercetin protects against atherosclerosis by regulating the expression of PCSK9, CD36, PPARγ, LXRα and ABCA1

Authors:
- Qingling Jia
- Hui Cao
- Dingzhu Shen
- Shanshan Li
- Li Yan
- Chuan Chen
- Sanli Xing
- Fangfang Dou
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Affiliations: Shanghai Geriatric Institute of Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 200031, P.R. China
Published online on: July 3, 2019 https://doi.org/10.3892/ijmm.2019.4263
Pages: 893-902
Copyright: © Jia et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The aim of this study was to investigate the mechanisms through which quercetin protects against atherosclerosis (AS) in apoE‑/‑ mice by regulating the expression of proprotein convertase subtilisin/kexin type 9 (PCSK9), cluster of differentiation 36 (CD36), peroxisome proliferator‑activated receptor γ (PPARγ), liver X receptor α (LXRα) and ATP binding cassette transporter A1 (ABCA1). We established an animal model of high‑fat diet induced AS using apoE‑/‑ mice. H&E, Oil Red O and Masson's trichrome staining were performed on aortic sinus and liver tissue sections to evaluate the histopathology, lipid accumulation and collagen deposition, respectively. Filipin staining was performed to detect free cholesterol (FC) in the aortic sinus. ELISA was performed to measure the serum levels of lipids including total cholesterol (TC), triglyceride (TG), high‑density lipoprotein‑cholesterol (HDL‑C), low‑density lipoprotein‑cholesterol (LDL‑C) and oxidized low‑density lipoprotein (oxLDL), as well as the levels of inflammatory cytokines, including tumor necrosis factor (TNF)‑α, interleukin (IL)‑6 and IL‑10. Western blot analysis was performed to analyze the protein expression levels of PCSK9, CD36, PPARγ, LXRα and ABCA1 in both the aorta and liver tissue. H&E staining revealed the presence of atherosclerotic plaques in the aortic sinus. Oil Red O staining revealed the existence of massive red‑stained lipids in the aortic sinus and Masson's trichrome staining revealed decreased collagen fibers and increased plaque instability. Filipin staining revealed that free cholesterol levels in the aorta sinus were increased. In addition, H&E staining suggested hepatocyte structural disorder in the model group, and Oil Red O staining revealed a cytoplasm filled with lipid droplets, which contained a large amount of red‑stained lipids. Masson's trichrome staining revealed that the liver tissue of the model group had fewer collagen fibers compared with that of the control group. Moreover, the mice in the model group had higher serum TC, LDL‑C, oxLDL, TNF‑α and IL‑6 levels, and lower IL‑10 levels. The protein expression levels of PCSK9 and CD36 were increased, while those of PPARγ, LXRα and ABCA1 were decreased in the aortas and livers of the model group mice. However, treatment with quercetin attenuated all these effects. On the whole, these results demonstrate that quercetin prevents the development of AS in apoE‑/‑ mice by regulating the expression of PCSK9, CD36, PPARγ, LXRα and ABCA1.

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1	Wang HH, Garruti G, Liu M, Portincasa P and Wang DQ: Cholesterol and lipoprotein metabolism and atherosclerosis: Recent advances in reverse cholesterol transport. Ann Hepatol. 16(Suppl 1: S3-S105): pp. S27–S42. 2017, View Article : Google Scholar : PubMed/NCBI
2	Adorni MP, Cipollari E, Favari E, Zanotti I, Zimetti F, Corsini A, Ricci C, Bernini F and Ferri N: Inhibitory effect of PCSK9 on Abca1 protein expression and cholesterol efflux in macrophages. Atherosclerosis. 256:1–6. 2017. View Article : Google Scholar
3	Chávez-Sánchez L, Garza-Reyes MG, Espinosa-Luna JE, Chávez-Rueda K, Legorreta-Haquet MV and Blanco-Favela F: The role of TLR2, TLR4 and CD36 in macrophage activation and foam cell formation in response to oxLDL in humans. Hum Immunol. 75:322–329. 2014. View Article : Google Scholar : PubMed/NCBI
4	Ding Z, Liu S, Wang X, Theus S, Deng X, Fan Y, Zhou S and Mehta JL: PCSK9 regulates expression of scavenger receptors and ox-LDL uptake in macrophages. Cardiovasc Res. 114:1145–1153. 2018. View Article : Google Scholar : PubMed/NCBI
5	He XW, Yu D, Li WL, Zheng Z, Lv CL, Li C, Liu P, Xu CQ, Hu XF and Jin XP: Anti-atherosclerotic potential of baicalin mediated by promoting cholesterol efflux from macrophages via the PPARγ-LXRα-ABCA1/ABCG1 pathway. Biomed Pharmacother. 83:257–264. 2016. View Article : Google Scholar : PubMed/NCBI
6	Lara-Guzman OJ, Tabares-Guevara JH, Leon-Varela YM, Álvarez RM, Roldan M, Sierra JA, Londoño-Londoño JA and Ramirez-Pineda JR: Proatherogenic macrophage activities are targeted by the flavonoid quercetin. J Pharmacol Exp Ther. 343:296–306. 2012. View Article : Google Scholar : PubMed/NCBI
7	Li S, Cao H, Shen D, Jia Q, Chen C and Xing S: Quercetin protects against ox-LDL-induced injury via regulation of ABCAl, LXR-α and PCSK9 in RAW264.7 macrophages. Mol Med Rep. 18:799–806. 2018.PubMed/NCBI
8	Lee SM, Moon J, Cho Y, Chung JH and Shin MJ: Quercetin up-regulates expressions of peroxisome proliferator-activated receptor γ, liver X receptorα, and ATP binding cassette transporter A1 genes and increases cholesterol efflux in human macrophage cell line. Nutr Res. 33:136–143. 2013. View Article : Google Scholar : PubMed/NCBI
9	Tang JM and Chen ML: Medical Laboratory Zoology. China Press of Traditional Chinese Medicine; Beijing: pp. 276–278. pp. 2862012
10	Yue P, Chen Z, Nassir F, Bernal-Mizrachi C, Finck B, Azhar S and Abumrad NA: Enhanced hepatic apoA-I secretion and peripheral efflux of cholesterol and phospholipid in CD36 null mice. PLoS One. 5:e99062010. View Article : Google Scholar : PubMed/NCBI
11	Majdalawieh A and Ro HS: PPARgamma1 and LXRalpha face a new regulator of macrophage cholesterol homeostasis and inflammatory responsiveness, AEBP1. Nucl Recept Signal. 8:e0042010. View Article : Google Scholar : PubMed/NCBI
12	Breslow JL: Mouse models of atherosclerosis. Science. 272:685–688. 1996. View Article : Google Scholar : PubMed/NCBI
13	Martens FM, Rabelink TJ, op 't Roodt J, de Koning EJ and Visseren FL: TNF-alpha induces endothelial dysfunction in diabetic adults, an effect reversible by the PPAR-gamma agonist pioglitazone. Eur Heart J. 27:1605–1609. 2006. View Article : Google Scholar : PubMed/NCBI
14	Frisdal E, Lesnik P, Olivier M, Robillard P, Chapman MJ, Huby T, Guerin M and Le GW: Interleukin-6 protects human macrophages from cellular cholesterol accumulation and attenuates the proinflammatory response. J Biol Chem. 286:30926–30936. 2011. View Article : Google Scholar : PubMed/NCBI
15	Terkeltaub RA: IL-10: An 'immunologic scalpel' for atherosclerosis? Arterioscler Thromb Vasc Biol. 19:2823–2825. 1999. View Article : Google Scholar : PubMed/NCBI
16	Lan H, Pang L, Smith MM, Levitan D, Ding W, Liu L, Shan LX, Shah VV, Laverty M, Arreaza G, et al: Proprotein convertase subtilisin/kexin type 9 (PCSK9) affects gene expression pathways beyond cholesterol metabolism in liver cells. J Cell Physiol. 224:273–281. 2010.PubMed/NCBI
17	Herbert B, Patel D, Waddington SN, Eden ER, McAleenan A, Sun XM and Soutar AK: Increased secretion of lipoproteins in transgenic mice expressing human D374Y PCSK9 under physiological genetic control. Arterioscler Thromb Vasc Biol. 30:1333–1339. 2010. View Article : Google Scholar : PubMed/NCBI
18	Jänis MT, Tarasov K, Ta HX, Suoniemi M, Ekroos K, Hurme R, Lehtimäki T, Päivä H, Kleber ME, März W, et al: Beyond LDL-C lowering: Distinct molecular sphingolipids are good indicators of proprotein convertase subtilisin/kexin type 9 (PCSK9) deficiency. Atherosclerosis. 228:380–385. 2013. View Article : Google Scholar : PubMed/NCBI
19	Silverstein RL and Febbraio M: CD36, a scavenger receptor involved in immunity, metabolism, angiogenesis, and behavior. Sci Signal. 2:pp. re32009, View Article : Google Scholar : PubMed/NCBI
20	Yazgan B, Ustunsoy S, Karademir B and Kartal ON: CD36 as a biomarker of atherosclerosis. Free Radic Biol Med. 75(Suppl 1): pp. S102014, View Article : Google Scholar
21	Febbraio M, Guy E and Silverstein RL: Stem cell transplantation reveals that absence of macrophage CD36 is protective against atherosclerosis. Arterioscler Thromb Vasc Biol. 24:2333–2338. 2004. View Article : Google Scholar : PubMed/NCBI
22	Tang ZH, Chun-Yan WU, Xie M, Liu LS and Jiang ZS: Effects of PCSK9 siRNA on CD36, SR-A1 and SR-B1 expression in THP-1 derived macrophages. Acta Univ Med Nanjing (Nat Sci). 31:pp. 673–678. 2011
23	Zhao L, Varghese Z, Moorhead JF, Chen Y and Ruan XZ: CD36 and lipid metabolism in the evolution of atherosclerosis. Br Med Bull. 126:101–112. 2018. View Article : Google Scholar : PubMed/NCBI
24	Ikhlef S, Berrougui H, Kamtchueng SO and Khalil A: Paraoxonase 1-treated oxLDL promotes cholesterol efflux from macrophages by stimulating the PPARgamma-LXRalpha-ABCA1 pathway. FEBS Lett. 590:1614–1629. 2016. View Article : Google Scholar : PubMed/NCBI
25	Larrede S, Quinn CM, Jessup W, Frisdal E, Olivier M, Hsieh V, Kim MJ, Van Eck M, Couvert P, Carrie A, et al: Stimulation of cholesterol efflux by LXR agonists in cholesterol-loaded human macrophages is ABCA1-dependent but ABCG1-independent. Arterioscler Thromb Vasc Biol. 29:1930–1936. 2009. View Article : Google Scholar : PubMed/NCBI
26	Zhong Q, Zhao S, Yu B, Wang X, Matyal R, Li Y and Jiang Z: High-density lipoprotein increases the uptake of oxidized low density lipoprotein via PPARγ/CD36 pathway in inflammatory adipocytes. Int J Biol Sci. 11:256–265. 2015. View Article : Google Scholar :
27	Kong LL, Cui QJ and Yi HX: Measurement of quercetin, kaempferide, and isorhamnetin content in Semen Cuscutae by HPLC. Chin Tradit Herbal Drugs. 35:112–113. 2004.In Chinese.
28	Lv L, Zhu YM and Xu DM: Study on flavones from Taxillus chinensis (DC.) Danser and assay of its quercetin. Chinese Traditional Patent Medicine. 26:1046–1048. 2004.
29	Han JJ, Hao J, Kim CH, Hong JS, Ahn HY and Lee YS: Quercetin prevents cardiac hypertrophy induced by pressure overload in rats. J Vet Med Sci. 71:737–743. 2009. View Article : Google Scholar : PubMed/NCBI
30	Juźwiak S, Wójcicki J, Mokrzycki K, Marchlewicz M, Białecka M, Wenda-Rózewicka L, Gawrońska-Szklarz B and Droździk M: Effect of quercetin on experimental hyperlipidemia and atherosclerosis in rabbits. Pharmacol Rep. 57:604–609. 2015.
31	Bhaskar S, Sudhakaran PR and Helen A: Quercetin attenuates atherosclerotic inflammation and adhesion molecule expression by modulating TLR-NF-κB signaling pathway. Cell Immunol. 310:131–140. 2016. View Article : Google Scholar : PubMed/NCBI
32	Lu XL, Zhao CH, Yao XL and Zhang H: Quercetin attenuates high fructose feeding-induced atherosclerosis by suppressing inflammation and apoptosis via ROS-regulated PI3K/AKT signaling pathway. Biomed Pharmacother. 85:658–671. 2017. View Article : Google Scholar
33	Chen M: Effect of quercetin on ox-LDL-induced lipid accumulation and peroxidation in mouse macrophages. Chin J Pathophysiol. 29:1370–1374. 2013.In Chinese.
34	Sun L, Li E, Wang F, Wang T, Qin Z, Niu S and Qiu C: Quercetin increases macrophage cholesterol efflux to inhibit foam cell formation through activating PPARγ-ABCA1 pathway. Int J Clin Exp Pathol. 8:10854–10860. 2015.
35	Ren K, Jiang T and Zhao GJ: Quercetin induces the selective uptake of HDL-cholesterol via promoting SR-BI expression and the activation of the PPARγ/LXRα pathway. Food Funct. 9:624–635. 2018. View Article : Google Scholar : PubMed/NCBI