Insulin‑receptor substrate 1 protects against injury in endothelial cell models of ox‑LDL‑induced atherosclerosis by inhibiting ER stress/oxidative stress‑mediated apoptosis and activating the Akt/FoxO1 signaling pathway

Liu,Juan; Yi,Xu; Tao,Yuan; Wang,Yanjiang; Xu,Zhiqiang

doi:10.3892/ijmm.2020.4728

Insulin‑receptor substrate 1 protects against injury in endothelial cell models of ox‑LDL‑induced atherosclerosis by inhibiting ER stress/oxidative stress‑mediated apoptosis and activating the Akt/FoxO1 signaling pathway

Authors:
- Juan Liu
- Xu Yi
- Yuan Tao
- Yanjiang Wang
- Zhiqiang Xu
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Affiliations: Department of Neurology, Daping Hospital, Army Medical University, Chongqing 400042, P.R. China
Published online on: September 15, 2020 https://doi.org/10.3892/ijmm.2020.4728
Pages: 1671-1682
Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Oxidized low‑density lipoprotein (ox‑LDL)‑induced endothelial cell (EC) injury is a risk factor for atherosclerosis. Therefore, the present study aimed to investigate the effects of insulin‑receptor substrate 1 (IRS‑1) on injury to ox‑LDL‑exposed ECs. For this purpose, thoracic aorta tissues were isolated from rats and cultured to obtain ECs, which were then identified using immunohistochemical staining. IRS‑1 overexpression plasmid (pcDNA3.1‑IRS‑1) and IRS‑1‑small interfering RNA were synthesized and transfected into ECs pre‑exposed to ox‑LDL. MTT and TUNEL assays were performed to evaluate the cell proliferative activity and apoptosis. Intracellular reactive oxygen species (ROS) production was determined by a flow cytometry assay. Reverse transcription‑quantitative PCR was conducted to measure the peroxisome proliferator‑activated receptor gamma co‑activator 1 alpha (Ppargcla), phosphoenolpyruvate carboxykinase 1 (Pck1) and glucose‑6‑phosphatase catalytic subunit (G6pc) gene transcription levels. Western blot analysis was then performed to determine the IRS‑1, forkhead box O1 (FoxO1), phosphorylated (p‑)FoxO1, 78‑kDa glucose‑regulated protein (GRP78), p‑eukaryotic translation initiation factor 2A (eIF2α), CHOP, Akt and p‑Akt expression levels. Immunofluorescence staining was used to evaluate p‑FoxO1 nuclear localization. The results indicated that IRS‑1 significantly enhanced the proliferative activity, whereas it inhibited the apoptosis of ECs in a model of ox‑LDL‑induced atherosclerosis compared with ECs without IRS‑1 treatment (P<0.05). IRS‑1 significantly decreased the p‑FoxO1/FoxO1 ratio compared with ECs without ox‑LDL treatment (P<0.05). IRS‑1 significantly downregulated the expression of ER stress biomarkers, including GRP78, CHOP and the p‑eIF2α/eIF2α ratio in ox‑LDL‑exposed ECs compared with ECs without ISR‑1 treatment (P<0.05). IRS‑1 significantly reduced the intracellular ROS levels in the EC models of ox‑LDL‑induced atherosclerosis compared with ECs without IRS‑1 treatment (P<0.05). Moreover, IRS‑1 promoted the phosphorylation of Akt in the EC models of ox‑LDL‑induced atherosclerosis. IRS‑1 also significantly suppressed the transcription of atherosclerosis‑associated genes in ox‑LDL‑exposed ECs compared with ECs without IRS‑1 treatment (P<0.05). Furthermore, IRS‑1 significantly increased the cytoplasmic localization of p‑FoxO1 in EC models of ox‑LDL‑induced atherosclerosis. On the whole, the findings of the present study demonstrate that IRS‑1 exerts protective effects in an EC model of ox‑LDL‑induced atherosclerosis by inhibiting ER stress/oxidative stress‑mediated apoptosis and activating the Akt/FoxO1 signaling pathway.

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1	Kishore SP, Blank E, Heller DJ, Patel A, Peters A, Price M, Vidula M, Fuster V, Onuma O, Huffman MD and Vedanthan R: Modernizing the world health organization list of essential medicines for preventing and controlling cardiovascular diseases. J Am Coll Cardiol. 71:564–574. 2018. View Article : Google Scholar : PubMed/NCBI
2	Wu Y, Benjamin EJ and MacMahon S: Prevention and control of cardiovascular disease in the rapidly changing economy of China. Circulation. 133:2545–2560. 2016. View Article : Google Scholar : PubMed/NCBI
3	Chen L, Yang W, Guo Y, Chen W, Zheng P, Zeng J and Tong W: Exosomal lncRNA GAS5 regulates the apoptosis of macrophages and vascular endothelial cells in atherosclerosis. PLoS One. 12:e01854062017. View Article : Google Scholar : PubMed/NCBI
4	Bano A, Chaker L, Mattace-Raso FU, van der Lugt A, Ikram MA, Franco OH, Peeters RP and Kavousi M: Thyroid function and the risk of atherosclerotic cardiovascular morbidtiy and mortality: The rotterdam study. Circ Res. 121:1392–1400. 2017. View Article : Google Scholar : PubMed/NCBI
5	Lusis AJ: Atherosclerosis. Nature. 407:233–241. 2000. View Article : Google Scholar : PubMed/NCBI
6	Novák J, Bienertová-Vašků J, Kára T and Novák M: MicroRNA involved in the lipid metabolism and their possible implications for atherosclerosis development and treatment. Mediators Inflamm. 2014:2758672014. View Article : Google Scholar
7	Takahashi Y, Zhu H and Yoshimoto T: Essential roles of lipoxygenases in LDL oxidation and development of atherosclerosis. Antioxid Redox Signal. 7:425–431. 2005. View Article : Google Scholar : PubMed/NCBI
8	Zi Y, Yi-An Y, Bing J, Yan L, Jing T, Chun-Yu G, Fan P, Hao L, Jia-Ni T, Han-Jin H, et al: Sirt6-induced autophagy restricted TREM-1 mediated pyroptosis with ox-LDL treated endothelial cells: Relevance to prognostication of patients with acute myocardial infarction. Cell Death Discov. 5:882019. View Article : Google Scholar
9	Mitchell JD, Fergestrom N, Cage BF, Paisley R, Moon P, Novak E, Cheezum M, Shaw LJ and Villines TC: Impact of Statins on cardiovascular outcomes following coronary artery calcium scoring. J Am Coll Cardiol. 72:3233–3242. 2018. View Article : Google Scholar : PubMed/NCBI
10	Elmore S: Apoptosis: A review of programmed cell death. Toxicol Pathol. 35:495–516. 2007. View Article : Google Scholar : PubMed/NCBI
11	Favaloro B, Allocati N, Graziano V, Di Ilio C and De Laurenzi V: Role of apoptosis of disease. Aging (Albany NY). 4:330–349. 2012.
12	Wei DH, Jia XY, Liu YH, Guo FX, Tang ZH, Li XH, Wang Z, Liu LS, Wang GX, Jian ZS and Ruan CG: Cathepsin L stimulates autophagy and inhibits apoptosis of ox-LDL induced endothelial cells: Potential role in atherosclerosis. Int J Mol Med. 31:400–406. 2013.
13	Hong D, Bai YP, Gao HC, Wang X, Li LF, Zhang GG and Hu CP: Ox-LDL induces endothelial cell apoptosis via the LOX-1-dependent endoplasmic reticulum stress pathway. Atherosclerosis. 235:310–317. 2014.PubMed/NCBI
14	Zhang D, Zhang X, Liu D, Liu T, Cai W, Yan C and Han Y: Association between insulin receptor substrate 1 polymorhpisms and high platelet reactivity with clopidogrel therapy in coronary artery disease patients with type 2 diabetes mellitus. Cardiovasc Diabetol. 15:502016.
15	Park K, Mima A, Li Q, Rask-Madsen C, He P, Mizutani K, Katagiri S, Maeda Y, Wu IH, Khamaisi M, et al: Insulin decreases atherosclerosis by inducing endothelin receptor B expression. JCI Insight. 1:e865742016.PubMed/NCBI
16	Galkina EV, Butcher M, Keller SR, Goff M, Bruce A, Pei H, Sarembock IJ, Sanders JM, Nagelin MH, Srinivasan S, et al: Accelerated atherosclerosis in apoe-/- mice heterozygous for the insulin receptor and the insulin receptor substrate-1. Arterioscler Thromb Vasc Biol. 32:247–256. 2012.
17	Xi G, Wai C, White MF and Clemmons DR: Down-Regulation of insulin receptor substrate 1 during hyperglycemia induces vascular smooth muscle cell dedifferentiation. J Biol Chem. 292:2009–2020. 2017.
18	Kedenko L, Lamina C, Kedenko I, Kollerits B, Kiesslich R, Iglseder B, Kronenberg F and Paulweber B: Genetic polymorphisms at SIRT1 and FoxO1 are associated with carotid atherosclerosis in the SAPHIR cohort. BMC Med Genet. 15:1122014.PubMed/NCBI
19	Luo Y, Meng X, Zhou P, Lu S, Qin M, Xu X, Sun G and Sun X: Elatoside C protects against ox-LDL induced HUVECs injury by FoxO1-mediated autophagy induction. Biochim Biophys Acta Mol Basis Dis. 1863:1654–1665. 2017.PubMed/NCBI
20	Piao X, Liu B, Sui X, Li S, Niu W, Zhang Q, Shi X, Cai S and Fan Y: Picroside II improves severe acute pancreatitis-induced intestinal barrier injury by inactivating oxidative and inflammatory TLR4-dependent PI3K/AKT/NF-κB signaling and improving gut microbiota. Oxid Med Cell Longev. 2020:35894972020.
21	Zhong X, Ma X, Zhang L, Li Y, Li Y and He R: MIAT promotes proliferation and hinders apoptosis by modulating miR-181b/STAT3 axis in ox-LDL-induced atherosclerosis cell models. Biomed Pharmacother. 97:1078–1085. 2018. View Article : Google Scholar
22	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
23	Chen J, Mehta JL, Haider N, Zhang X, Narula J and Li D: Role of caspases in ox-LDL-induced apoptotic cascade in human coronary artery endothelial cells. Circ Res. 94:370–376. 2004. View Article : Google Scholar
24	Qin M, Luo Y, Meng Xb, Wang M, Wang Hw, Song Sy, Ye Jx, Pan Rl, Yao F, Wu P, et al: Myricitrin attenuates endothelial cell apoptosis to prevent atherosclerosis: An insight into PI3K/Akt activation and STAT3 signaling pathway. Vascul Pharmacol. 70:23–34. 2015. View Article : Google Scholar : PubMed/NCBI
25	Cahill-Smith S and Li JM: Oxidative stress, redox signalling and endothelial dysfunction of ageing-related neurodegenerative diseases: A role of NADPH oxidase 2. Br J Clin Pharmacol. 78:441–453. 2014. View Article : Google Scholar : PubMed/NCBI
26	Xue Z, Yuan W, Li J, Zhou H, Xu L, Weng J, Li X, Zhang X, Wang Z and Yan J: Cyclophilin A mediates the ox-LDL induced activation and apoptosis of macrophages with autophagy. Int J Cardiol. 230:142–148. 2017. View Article : Google Scholar : PubMed/NCBI
27	Han X, Han X, Wang Z, Shen J and Dong Q: HDAC9 regulates ox-LDL-induced endothelial apoptosis by participating in inflammatory reactions. Front Biosci (Landmark Ed). 21:907–17. 2016. View Article : Google Scholar
28	Glab JA, Doerflinger M, Nedeva C, Jose I, Mbogo GW, Paton JC, Paton AW, Kueh AJ, Herold MJ, Huang DC, et al: DR5 and caspase 8 are dispensable in ER stress induced apoptosis. Cell Death Differ. 24:944–950. 2017. View Article : Google Scholar : PubMed/NCBI
29	Kishino A, Hayashi K, Hidai C, Masuda T, Nomura Y and Oshima T: XBP1-FoxO1 interaction regulates ER stress induced autophagy in auditory cells. Sci Rep. 7:44422017. View Article : Google Scholar
30	Tanaka J, Qiang L, Banks AS, Welch CL, Matsumoto M, Kitamura T, Ido-Kitamura Y, DePinho RA and Accili D: FoxO1 links hyperglycemia to LDL oxidation and endothelial nitric oxide synthase dysfunction in vascular endothelial cells. Diabetes. 58:2344–2354. 2009. View Article : Google Scholar : PubMed/NCBI
31	Li Y, Ren M, Wang X, Cui X, Zhao H, Zhao C, Zhou J, Guo Y, Hu Y, Yan C, et al: Glutaredoxin 1 mediates the protective effect of steady laminar flow on endotelial cells against oxidative stress-induced apoptosis via inhibiting bim. Sci Rep. 7:155392017. View Article : Google Scholar