Pro‑atherogenic activation of A7r5 cells induced by the oxLDL/β2GPI/anti‑β2GPI complex

Wang,Ting; Ouyang,Hang; Zhou,Hong; Xia,Longfei; Wang,Xiaoyan; Wang,Ting

doi:10.3892/ijmm.2018.3805

Pro‑atherogenic activation of A7r5 cells induced by the oxLDL/β2GPI/anti‑β2GPI complex

Authors:
- Ting Wang
- Hang Ouyang
- Hong Zhou
- Longfei Xia
- Xiaoyan Wang
View Affiliations

Affiliations: Jiangsu Key Laboratory of Medicine Science and Laboratory Medicine, School of Medicine, Jiangsu University, Zhenjiang, Jiangsu 212013, P.R. China
Published online on: August 2, 2018 https://doi.org/10.3892/ijmm.2018.3805
Pages: 1955-1966
Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

A previous study has revealed that oxidized low‑density lipoprotein (oxLDL)/β2‑glycoprotein I (β2GPI)/anti‑β2‑glycoprotein I (anti‑β2GPI), an immune complex, is able to activate the Toll‑like receptor 4 (TLR4)/nuclear factor κβ (NF‑κβ) inflammatory signaling pathway in macrophages, and consequently enhance foam cell formation and the secretion of prothrombin activators. However, the effects of the oxLDL/β2GPI/anti‑β2GPI complex on vascular smooth muscle cells have yet to be investigated. The present study investigated whether the oxLDL/β2GPI/anti‑β2GPI complex was able to reinforce the pro‑atherogenic activities of a rat thoracic aorta smooth muscle cell line (A7r5) and examined the underlying molecular mechanisms. The results revealed that the oxLDL/β2GPI/anti‑β2GPI complex treatment significantly (P<0.05 vs. the media, oxLDL, oxLDL/β2GPI and β2GPI/anti‑β2GPI groups) enhanced the pro‑atherogenic activation of A7r5 cells, including intracellular lipid loading, Acyl‑coenzyme A cholesterol acyltransferase mRNA expression, migration, matrix metalloproteinase‑9 and monocyte chemoattractant protein 1 secretion, all via TLR4. In addition, the expression of TLR4 and the phosphorylation of NF‑κβ p65, p38 and ERK1/2 were also upregulated in oxLDL/β2GPI/anti‑β2GPI complex‑treated A7r5 cells. Pre‑treatment with TAK‑242, a TLR4 inhibitor, was able to partly attenuate the oxLDL/β2GPI/anti‑β2GPI complex‑induced phosphorylation of NF‑κβ p65; however, it had no effect on the phosphorylation of extracellular regulated kinase 1/2 (ERK1/2) and p38. Meanwhile, the NF‑κβ p65 inhibitor ammonium pyrrolidinedithiocarbamate and the ERK1/2 inhibitor U0126, but not the p38 inhibitor SB203580, were able to block oxLDL/β2GPI/anti‑β2GPI complex‑induced foam cell formation and migration in A7r5 cells. Hence, it was demonstrated that the oxLDL/β2GPI/anti‑β2GPI complex is able to enhance the lipid uptake, migration and active molecule secretion of A7r5 cells via TLR4, and finally deteriorate atherosclerosis plaques. Additionally, it was demonstrated that oxLDL/β2GPI/anti‑β2GPI complex‑induced foam cell formation and migration may be partly mediated by the TLR4/NF‑κβ signaling pathway and that ERK1/2 may also participate in the process.

View Figures

View References

1	Glass CK and Witztum JL: Atherosclerosis. the road ahead. Cell. 104:503–516. 2001. View Article : Google Scholar : PubMed/NCBI
2	Amaya-Amaya J, Rojas-Villarraga A and Anaya JM: Cardiovascular disease in the antiphospholipid syndrome. Lupus. 23:1288–1291. 2014. View Article : Google Scholar : PubMed/NCBI
3	Matsuura E, Atzeni F, Sarzi-Puttini P, Turiel M, Lopez LR and Nurmohamed MT: Is atherosclerosis an autoimmune disease? BMC Med. 12:472014. View Article : Google Scholar : PubMed/NCBI
4	Stern C, Chamley L, Hale L, Kloss M, Speirs A and Baker HW: Antibodies to beta2 glycoprotein I are associated with in vitro fertilization implantation failure as well as recurrent miscarriage: Results of a prevalence study. Fertil Steril. 70:938–944. 1998. View Article : Google Scholar : PubMed/NCBI
5	Habe K, Wada H, Matsumoto T, Ohishi K, Ikejiri M, Matsubara K, Morioka T, Kamimoto Y, Ikeda T, Katayama N, et al: Presence of antiphospholipid antibody is a risk factor in thrombotic events in patients with antiphospholipid syndrome or relevant diseases. Intern Med. 97:345–350. 2013.
6	Amory CF, Levine SR, Brey RL, Gebregziabher M, Tuhrim S, Tilley BC, Simpson AC, Sacco RL and Mohr JP; APASS-WARSS Collaborators: Antiphospholipid antibodies and recurrent thrombotic events: Persistence and portfolio. Cerebrovasc Dis. 40:293–300. 2015. View Article : Google Scholar : PubMed/NCBI
7	Li J, Chi Y, Liu S, Wang L, Wang R, Han X, Matsuura E and Liu Q: Recombinant domain V of β₂-glycoprotein I inhibits the formation of atherogenic oxLDL/β₂-glycoprotein I complexes. J Clin Immunol. 34:669–676. 2014. View Article : Google Scholar : PubMed/NCBI
8	Otomo K, Amengual O, Fujieda Y, Nakagawa H, Kato M, Oku K, Horita T, Yasuda S, Matsumoto M, Nakayama KI, et al: Role of apolipoprotein B100 and oxidized low-density lipoprotein in the monocyte tissue factor induction mediated by anti-β₂ glyco-protein I antibodies. Lupus. 25:1288–1298. 2016. View Article : Google Scholar : PubMed/NCBI
9	Ouweneel AB and Van Eck M: Lipoproteins as modulators of atherothrombosis, From endothelial function to primary and secondary coagulation. Vascul Pharmacol. 82:1–10. 2016. View Article : Google Scholar
10	Di Pietro N, Formoso G and Pandolfi A: Physiology and pathophysiology of oxLDL uptake by vascular wall cells in atherosclerosis. Vascul Pharmacol. 84:1–7. 2016. View Article : Google Scholar : PubMed/NCBI
11	George J, Harats D, Gilburd B, Afek A, Levy Y, Schneiderman J, Barshack I, Kopolovic J and Shoenfeld Y: Immunolocalization of beta2-glycoprotein I (apolipoprotein H) to human athero-sclerotic plaques: Potential implications for lesion progression. Circulation. 99:2227–2230. 1999. View Article : Google Scholar : PubMed/NCBI
12	Yu R, Yuan Y, Niu D, Song J, Liu T, Wu J and Wang J: Elevated beta2-glycoprotein I-low-density lipoprotein levels are associated with the presence of diabetic microvascular complications. J Diabetes Complications. 29:59–63. 2015. View Article : Google Scholar
13	Kasahara J, Kobayashi K, Maeshima Y, Yamasaki Y, Yasuda T, Matsuura E and Makino H: Clinical significance of serum oxidized low-density lipoprotein/beta2-glycoprotein I complexes in patients with chronic renal diseases. Nephron Clin Pract. 98:c15–c24. 2004. View Article : Google Scholar : PubMed/NCBI
14	Lopez D, Garcia-Valladares I, Palafox-Sanchez CA, De La Torre IG, Kobayashi K, Matsuura E and Lopez LR: Oxidized low-density lipoprotein/beta2-glycoprotein I complexes and autoantibodies to oxLig-1/beta2-glycoprotein I in patients with systemic lupus erythematosus and antiphospholipid syndrome. Am J Clin Pathol. 121:426–436. 2004. View Article : Google Scholar : PubMed/NCBI
15	Matsuura E and Lopez LR: Are oxidized LDL/beta2-glycoprotein I complexes pathogenic antigens in autoimmune-mediated atherosclerosis? Clin Dev Immunol. 11:103–111. 2004. View Article : Google Scholar : PubMed/NCBI
16	Kobayashi K, Kishi M, Atsumi T, Bertolaccini ML, Makino H, Sakairi N, Yamamoto I, Yasuda T, Khamashta MA, Hughes GR, et al: Circulating oxidized LDL forms complexes with beta2-glycoprotein I: Implication as an atherogenic auto-antigen. J Lipid Res. 44:716–726. 2003. View Article : Google Scholar : PubMed/NCBI
17	Hasunuma Y, Matsuura E, Makita Z, Katahira T, Nishi S and Koike T: Involvement of beta 2-glycoprotein I and anticardiolipin antibodies in oxidatively modified low-density lipoprotein uptake by macrophages. Clin Exp Immunol. 107:569–573. 1997. View Article : Google Scholar : PubMed/NCBI
18	Kobayashi K, Tada K, Itabe H, Ueno T, Liu PH, Tsutsumi A, Kuwana M, Yasuda T, Shoenfeld Y, de Groot PG and Matsuura E: Distinguished effects of antiphospholipid antibodies and anti-oxidized LDL antibodies on oxidized LDL uptake by macrophages. Lupus. 16:929–938. 2007. View Article : Google Scholar : PubMed/NCBI
19	Xu Y, Kong X, Zhou H, Zhang X, Liu J, Yan J, Xie H and Xie Y: oxLDL/β₂GPI/anti-β₂GPI complex induced macrophage differentiation to foam cell involving TLR4/NF-kappa B signal transduction pathway. Thromb Res. 134:384–392. 2014. View Article : Google Scholar : PubMed/NCBI
20	Bassi N, Ghirardello A, Iaccarino L, Zampieri S, Rampudda ME, Atzeni F, Sarzi-Puttini P, Shoenfeld Y and Doria A: OxLDL/beta2GPI-anti-oxLDL/beta2GPI complex and atherosclerosis in SLE patients. Autoimmun Rev. 7:52–58. 2007. View Article : Google Scholar : PubMed/NCBI
21	Zhang X, Xie Y, Zhou H, Xu Y, Liu J, Xie H and Yan J: Involvement of TLR4 in oxidized LDL/β₂GPI/anti-β₂GPI-induced transformation of macrophages to foam cells. J Atheroscler Thromb. 21:1140–1151. 2014. View Article : Google Scholar
22	Tabas I, García-Cardeña G and Owens GK: Recent insights into the cellular biology of atherosclerosis. J Cell Biol. 209:13–22. 2015. View Article : Google Scholar : PubMed/NCBI
23	Bennett MR, Sinha S and Owens GK: Vascular smooth muscle cells in atherosclerosis. Circ Res. 118:692–702. 2016. View Article : Google Scholar : PubMed/NCBI
24	Chistiakov DA, Orekhov AN and Bobryshev YV: Vascular smooth muscle cell in atherosclerosis. Acta Physiol(Oxf). 214:33–50. 2015. View Article : Google Scholar
25	Hartman J and Frishman WH: Inflammation and atherosclerosis: A review of the role of interleukin-6 in the development of atherosclerosis and the potential for targeted drug therapy. Cardiol Rev. 22:147–151. 2014. View Article : Google Scholar : PubMed/NCBI
26	Matsuura E, Lopez LR, Shoenfeld Y and Ames PR: β₂-glycoprotein I and oxidative inflammation in early athero-genesis: A progression from innate to adaptive immunity? Autoimmun Rev. 12:241–249. 2012. View Article : Google Scholar : PubMed/NCBI
27	Roshan MH and Tambo A: The role of TLR2, TLR4, and TLR9 in the pathogenesis of atherosclerosis. Int J Inflam. 2016:15328322016. View Article : Google Scholar : PubMed/NCBI
28	Hopkins PN: Molecular biology of atherosclerosis. Physiol Rev. 93:1317–1542. 2013. View Article : Google Scholar : PubMed/NCBI
29	Yin YW, Liao SQ, Zhang MJ, Liu Y, Li BH, Zhou Y, Chen L, Gao CY, Li JC and Zhang LL: TLR4-mediated inflammation promotes foam cell formation of vascular smooth muscle cell by upregulating ACAT1 expression. Cell Death Dis. 6:16592015. View Article : Google Scholar : PubMed/NCBI
30	Yang K, Zhang XJ, Cao LJ, Liu XH, Liu ZH, Wang XQ, Chen QJ, Lu L, Shen WF and Liu Y: Toll-like receptor 4 mediates inflammatory cytokine secretion in smooth muscle cells induced by oxidized low-density lipoprotein. PloS One. 9:e959352014. View Article : Google Scholar : PubMed/NCBI
31	Kong F, Ye B, Cao J, Cai X, Lin L, Huang S, Huang W and Huang Z: Curcumin represses NLRP3 inflammasome activation via TLR4/MyD88/NF-κB and P2X7R signaling in PMA-induced macrophages. Front Pharmacol. 7:3692016. View Article : Google Scholar
32	Jing Q, Xin SM, Cheng ZJ, Zhang WB, Zhang R, Qin YW and Pei G: Activation of p38 mitogen-activated protein kinase by oxidized LDL in vascular smooth muscle cells: Mediation via pertussis toxin-sensitive g proteins and association with oxidized LDL-induced cytotoxicity. Circ Res. 84:831–839. 1999. View Article : Google Scholar : PubMed/NCBI
33	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar
34	Newby AC: Matrix metalloproteinases regulate migration, proliferation, and death of vascular smooth muscle cells by degrading matrix and non-matrix substrates. Cardiovasc Res. 69:614–624. 2006. View Article : Google Scholar
35	Lin J, Kakkar V and Lu X: Impact of MCP-1 in atherosclerosis. Curr Pharm Des. 20:4580–4588. 2014. View Article : Google Scholar : PubMed/NCBI
36	Płóciennikowska A, Hromada-Judycka A, Borzęcka K and Kwiatkowska K: Co-operation of TLR4 and raft proteins in LPS-induced pro-inflammatory signaling. Cell Mol Life Sci. 72:557–581. 2015. View Article : Google Scholar
37	Ross R and Glomset JA: Atherosclerosis and the arterial smooth muscle cell: Proliferation of smooth muscle is a key event in the genesis of the lesions of atherosclerosis. Science. 180:1332–1339. 1973. View Article : Google Scholar : PubMed/NCBI
38	Badimon L and Vilahur G: Thrombosis formation on athero-sclerotic lesions and plaque rupture. J Intern Med. 276:618–632. 2014. View Article : Google Scholar : PubMed/NCBI
39	Sima P, Vannucci L and Vetvicka V: Atherosclerosis as autoimmune disease. Ann Transl Med. 6:1162018. View Article : Google Scholar : PubMed/NCBI
40	Tanaka N, Masuoka S, Kusunoki N, Nanki T and Kawai S: Serum resistin level and progression of atherosclerosis during glucocor-ticoid therapy for systemic autoimmune diseases. Metabolites. 6:E282016. View Article : Google Scholar
41	Rosenfeld ME and Ross R: Macrophage and smooth muscle cell proliferation in atherosclerotic lesions of WHHL and comparably hypercholesterolemic fat-fed rabbits. Arteriosclerosis. 10:680–687. 1990. View Article : Google Scholar : PubMed/NCBI
42	Suckling KE and Strange EF: Role of acyl-CoA: Cholesterol acyltransferase in cellular cholesterol metabolism. J Lipid Res. 26:647–671. 1985.PubMed/NCBI
43	Rudel LL, Lee RG and Cockman TL: Acyl coenzyme A: Cholesterol acyltransferase types 1 and 2: Structure and function in atherosclerosis. Curr Opin Lipidol. 12:121–127. 2001. View Article : Google Scholar : PubMed/NCBI
44	Sakashita N, Miyazaki A, Takeya M, Horiuchi S, Chang CC, Chang TY and Takahashi K: Localization of human acyl-coenzyme A: Cholesterol acyltransferase-1 (ACAT-1) in macrophages and in various tissues. Am J Pathol. 156:227–236. 2000. View Article : Google Scholar : PubMed/NCBI
45	Dai Y, Cao Y, Zhang Z, Vallurupalli S and Mehta JL: Xanthine oxidase induces foam cell formation through LOX-1 and NLRP3 activation. Cardiovasc Drugs Ther. 31:19–27. 2017. View Article : Google Scholar : PubMed/NCBI
46	Yang J, Jian R, Yu J, Zhi X, Liao X, Yu J and Zhou P: CD73 regulates vascular smooth muscle cell functions and facilitates atherosclerotic plaque formation. IUBMB Life. 67:853–860. 2015. View Article : Google Scholar : PubMed/NCBI
47	Kim J and Ko J: Human sLZIP promotes atherosclerosis via MMP-9 transcription and vascular smooth muscle cell migration. FASEB J. 28:5010–5021. 2014. View Article : Google Scholar : PubMed/NCBI
48	Chae IG, Yu MH, Im NK, Jung YT, Lee J, Chun KS and Lee IS: Effect of Rosemarinus officinalis L. on MMP-9, MCP-1 levels, and cell migration in RAW 264.7 and smooth muscle cells. J Med Food. 15:879–886. 2012. View Article : Google Scholar : PubMed/NCBI
49	Xie HX, Zhou H, Wang HB, Chen DD, Xia LF, Wang T and Yan JC: Anti-β(2)GPI/β(2)GPI induced TF and TNF-α expression in monocytes involving both TLR4/MyD88 and TLR4/TRIF signaling pathways. Mol Immunol. 53:246–254. 2013. View Article : Google Scholar
50	Cheng YC, Sheen JM, Hu WL and Hung YC: Polyphenols and oxidative stress in atherosclerosis-related ischemic heart disease and stroke. Oxid Med Cell Longev. 2017:85264382017. View Article : Google Scholar
51	Chaabane C, Coen M and Bochaton-Piallat ML: Smooth muscle cell phenotypic switch: implications for foam cell formation. Curr Opin Lipidol. 25:374–379. 2014. View Article : Google Scholar : PubMed/NCBI