Abstract
The study aims to explore the fluctuating expression of C/EBP Homologous Protein (CHOP) following rat carotid artery injury and its central role in vascular stenosis. Using in vivo rat carotid artery injury models and in vitro ischemia and hypoxia cell models employing human aortic endothelial cells (HAECs) and vascular smooth muscle cells (T/G HA-VSMCs), a comprehensive investigative framework was established. Histological analysis confirmed intimal hyperplasia in rat models. CHOP expression in vascular tissues was assessed using Western blot and immunohistochemical staining, and its presence in HAECs and T/G HA-VSMCs was determined through RT-PCR and Western blot. The study evaluated HAEC apoptosis, inflammatory cytokine secretion, cell proliferation, and T/G HA-VSMCs migration through Western blot, ELISA, CCK8, and Transwell migration assays. The rat carotid artery injury model revealed substantial fibrous plaque formation and vascular stenosis, resulting in an increased intimal area and plaque-to-lumen area ratio. Notably, CHOP is markedly elevated in vessels of the carotid artery injury model compared to normal vessels. Atorvastatin effectively mitigated vascular stenosis and suppresses CHOP protein expression. In HAECs, ischemia and hypoxia-induced CHOP upregulation, along with heightened TNFα, IL-6, caspase3, and caspase8 levels, while reducing cell proliferation. Atorvastatin demonstrated a dose-dependent suppression of CHOP expression in HAECs. Downregulation of CHOP or atorvastatin treatment led to reduced IL-6 and TNFα secretion, coupled with augmented cell proliferation. Similarly, ischemia and hypoxia conditions increased CHOP expression in T/G HA-VSMCs, which was concentration-dependently inhibited by atorvastatin. Furthermore, significantly increased MMP-9 and MMP-2 concentrations in the cell culture supernatant correlated with enhanced T/G HA-VSMCs migration. However, interventions targeting CHOP downregulation and atorvastatin usage curtailed MMP-9 and MMP-2 secretion and suppressed cell migration. In conclusion, CHOP plays a crucial role in endothelial injury, proliferation, and VSMCs migration during carotid artery injury, serving as a pivotal regulator in post-injury fibrous plaque formation and vascular remodeling. Statins emerge as protectors of endothelial cells, restraining VSMCs migration by modulating CHOP expression.
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Data Availability
The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.
References
Al-Khatib SM, Stevenson WG, Ackerman MJ, Bryant WJ, Callans DJ, Curtis AB, Deal BJ, Dickfeld T, Field ME, Fonarow GC, Gillis AM, Granger CB, Hammill SC, Hlatky MA, Joglar JA, Kay GN, Matlock DD, Myerburg RJ, Page RL (2018) 2017 AHA/ACC/HRS guideline for management of patients with ventricular arrhythmias and the prevention of sudden cardiac death: Executive summary: a report of the American College of Cardiology/American Heart Association Task Force on Clinical Practice Guidelines and the Heart Rhythm Society. Heart Rhythm 15:e190–e252. https://doi.org/10.1016/j.hrthm.2017.10.035
Aoki J, Tanabe K (2021) Mechanisms of drug-eluting stent restenosis. Cardiovasc Interv Ther 36:23–29. https://doi.org/10.1007/s12928-020-00734-7
Buccheri D, Piraino D, Andolina G, Cortese B (2016) Understanding and managing in-stent restenosis: a review of clinical data, from pathogenesis to treatment. J Thorac Dis 8:E1150–E1162. https://doi.org/10.21037/jtd.2016.10.93
Cheng WP, Wang BW, Shyu KG (2009) Regulation of GADD153 induced by mechanical stress in cardiomyocytes. Eur J Clin Invest 39:960–971. https://doi.org/10.1111/j.1365-2362.2009.02193.x
Chung IM, Gold HK, Schwartz SM, Ikari Y, Reidy MA, Wight TN (2002) Enhanced extracellular matrix accumulation in restenosis of coronary arteries after stent deployment. J Am Coll Cardiol 40:2072–2081. https://doi.org/10.1016/s0735-1097(02)02598-6
Clare J, Ganly J, Bursill CA, Sumer H, Kingshott P, de Haan JB (2022) The mechanisms of restenosis and relevance to next generation stent design. Biomolecules. https://doi.org/10.3390/biom12030430
Curcio A, Torella D, Indolfi C (2011) Mechanisms of smooth muscle cell proliferation and endothelial regeneration after vascular injury and stenting: approach to therapy. Circ J 75:1287–1296. https://doi.org/10.1253/circj.cj-11-0366
De Gregorio J, Aoki Y (2021) In-stent restenosis: burn and rebuild? Cardiovasc Revasc Med 22:50–51. https://doi.org/10.1016/j.carrev.2020.09.045
Demirtas K (2018) Inflammation and in-stent restenosis. Angiology 69:89. https://doi.org/10.1177/0003319717722103
Forstermann U, Xia N, Li H (2017) Roles of vascular oxidative stress and nitric oxide in the pathogenesis of atherosclerosis. Circ Res 120:713–735. https://doi.org/10.1161/CIRCRESAHA.116.309326
Gach O, Davin L, Lempereur M, Marechal P, Martinez C, Lancellotti P (2019) Percutaneous transluminal coronary angioplasty: from revolution to evolution. Rev Med Liege 74:S34–S38
Giustino G, Colombo A, Camaj A, Yasumura K, Mehran R, Stone GW, Kini A, Sharma SK (2022) Coronary In-stent restenosis: JACC state-of-the-art review. J Am Coll Cardiol 80:348–372. https://doi.org/10.1016/j.jacc.2022.05.017
Gori T (2022) Restenosis after coronary stent implantation: cellular mechanisms and potential of endothelial progenitor cells (a short guide for the interventional cardiologist). Cells. https://doi.org/10.3390/cells11132094
Guan G, Lei L, Lv Q, Gong Y, Yang L (2019) Curcumin attenuates palmitic acid-induced cell apoptosis by inhibiting endoplasmic reticulum stress in H9C2 cardiomyocytes. Hum Exp Toxicol 38:655–664. https://doi.org/10.1177/0960327119836222
Guo R, Wu Z, Jiang J, Liu C, Wu B, Li X, Li T, Mo H, He S, Li S, Yan H, Huang R, You Q, Wu K (2017) New mechanism of lipotoxicity in diabetic cardiomyopathy: deficiency of endogenous H(2)S production and ER stress. Mech Ageing Dev 162:46–52. https://doi.org/10.1016/j.mad.2016.11.005
Han K, Hassanzadeh S, Singh K, Menazza S, Nguyen TT, Stevens MV, Nguyen A, San H, Anderson SA, Lin Y, Zou J, Murphy E, Sack MN (2017) Parkin regulation of CHOP modulates susceptibility to cardiac endoplasmic reticulum stress. Sci Rep 7:2093. https://doi.org/10.1038/s41598-017-02339-2
Hu H, Tian M, Ding C, Yu S (2018) The C/EBP homologous Protein (CHOP) transcription factor functions in endoplasmic reticulum stress-induced apoptosis and microbial infection. Front Immunol 9:3083. https://doi.org/10.3389/fimmu.2018.03083
Juni RP, Duckers HJ, Vanhoutte PM, Virmani R, Moens AL (2013) Oxidative stress and pathological changes after coronary artery interventions. J Am Coll Cardiol 61:1471–1481. https://doi.org/10.1016/j.jacc.2012.11.068
Kawai K, Virmani R, Finn AV (2022) In-stent restenosis. Interv Cardiol Clin 11:429–443. https://doi.org/10.1016/j.iccl.2022.02.005
Kim TW, Lee SY, Kim M, Cheon C, Ko SG (2018) Kaempferol induces autophagic cell death via IRE1-JNK-CHOP pathway and inhibition of G9a in gastric cancer cells. Cell Death Dis 9:875. https://doi.org/10.1038/s41419-018-0930-1
Kumar A, Avishay DM, Jones CR, Shaikh JD, Kaur R, Aljadah M, Kichloo A, Shiwalkar N, Keshavamurthy S (2021) Sudden cardiac death: epidemiology, pathogenesis and management. Rev Cardiovasc Med 22:147–158. https://doi.org/10.31083/j.rcm.2021.01.207
Lan H, Wang Y, Yin T, Wang Y, Liu W, Zhang X, Yu Q, Wang Z, Wang G (2016) Progress and prospects of endothelial progenitor cell therapy in coronary stent implantation. J Biomed Mater Res B Appl Biomater 104:1237–1247. https://doi.org/10.1002/jbm.b.33398
Lawton JS, Tamis-Holland JE, Bangalore S, Bates ER, Beckie TM, Bischoff JM, Bittl JA, Cohen MG, DiMaio JM, Don CW, Fremes SE, Gaudino MF, Goldberger ZD, Grant MC, Jaswal JB, Kurlansky PA, Mehran R, Metkus TS Jr, Nnacheta LC, Rao SV, Sellke FW, Sharma G, Yong CM, Zwischenberger BA (2022) 2021 ACC/AHA/SCAI guideline for coronary artery revascularization: executive summary: a report of the American College of Cardiology/American Heart Association Joint Committee on Clinical Practice Guidelines. Circulation 145:e4–e17. https://doi.org/10.1161/CIR.0000000000001039
Lee MJ, Kwak YK, You KR, Lee BH, Kim DG (2009) Involvement of GADD153 and cardiac ankyrin repeat protein in cardiac ischemia-reperfusion injury. Exp Mol Med 41:243–252. https://doi.org/10.3858/emm.2009.41.4.027
Liao Y, Chen K, Dong X, Li W, Li G, Huang G, Song W, Chen L, Fang Y (2018) Berberine inhibits cardiac remodeling of heart failure after myocardial infarction by reducing myocardial cell apoptosis in rats. Exp Ther Med 16:2499–2505. https://doi.org/10.3892/etm.2018.6438
Liu W, Huang J, He S, Du R, Shi W, Wang Y, Du D, Du Y, Liu Q, Wang Y, Wang G, Yin T (2023a) Senescent endothelial cells’ response to the degradation of bioresorbable scaffold induces intimal dysfunction accelerating in-stent restenosis. Acta Biomater 166:266–277. https://doi.org/10.1016/j.actbio.2023.05.028
Liu L, Lan X, Chen X, Dai S, Wang Z, Zhao A, Lu L, Huang N, Chen J, Yang P, Liao Y (2023b) Multi-functional plant flavonoids regulate pathological microenvironments for vascular stent surface engineering. Acta Biomater 157:655–669. https://doi.org/10.1016/j.actbio.2022.11.035
Loinard C, Zouggari Y, Rueda P, Ramkhelawon B, Cochain C, Vilar J, Recalde A, Richart A, Charue D, Duriez M, Mori M, Arenzana-Seisdedos F, Levy BI, Heymes C, Silvestre JS (2012) C/EBP homologous protein-10 (CHOP-10) limits postnatal neovascularization through control of endothelial nitric oxide synthase gene expression. Circulation 125:1014–1026. https://doi.org/10.1161/CIRCULATIONAHA.111.041830
Moscarella E, Varricchio A, Stabile E, Franzone A, Granata F, Rapacciuolo A, Galasso G, Capozzolo C, Cirillo P, Monda V, Monterforte I, Borgia F, Piro O, Bonzani G, Trimarco B, Esposito G (2015) Bioabsorbable drug-eluting vascular scaffold for the treatment of coronary in-stent restenosis: a two center registry. Cardiovasc Revasc Med 16:401–405. https://doi.org/10.1016/j.carrev.2015.07.008
Oyadomari S, Mori M (2004) Roles of CHOP/GADD153 in endoplasmic reticulum stress. Cell Death Differ 11:381–389. https://doi.org/10.1038/sj.cdd.4401373
Panchatcharam M, Miriyala S, Yang F, Leitges M, Chrzanowska-Wodnicka M, Quilliam LA, Anaya P, Morris AJ, Smyth SS (2010) Enhanced proliferation and migration of vascular smooth muscle cells in response to vascular injury under hyperglycemic conditions is controlled by beta3 integrin signaling. Int J Biochem Cell Biol 42:965–974. https://doi.org/10.1016/j.biocel.2010.02.009
Ren JL, Chen Y, Zhang LS, Zhang YR, Liu SM, Yu YR, Jia MZ, Tang CS, Qi YF, Lu WW (2021) Intermedin(1–53) attenuates atherosclerotic plaque vulnerability by inhibiting CHOP-mediated apoptosis and inflammasome in macrophages. Cell Death Dis 12:436. https://doi.org/10.1038/s41419-021-03712-w
Rozpedek W, Pytel D, Mucha B, Leszczynska H, Diehl JA, Majsterek I (2016) The role of the PERK/eIF2alpha/ATF4/CHOP signaling pathway in tumor progression during endoplasmic reticulum stress. Curr Mol Med 16:533–544. https://doi.org/10.2174/1566524016666160523143937
Sallustro M, Peluso A, Turchino D, Maione I, Vita F, Martelli E, Serra R, Bracale UM (2023) Results of new dual-drug coated balloon angioplasty versus POBA for femoropopliteal lesions. Ann Vasc Surg 89:52–59. https://doi.org/10.1016/j.avsg.2022.09.047
Scull CM, Tabas I (2011) Mechanisms of ER stress-induced apoptosis in atherosclerosis. Arterioscler Thromb Vasc Biol 31:2792–2797. https://doi.org/10.1161/ATVBAHA.111.224881
Shaito A, Aramouni K, Assaf R, Parenti A, Orekhov A, Yazbi AE, Pintus G, Eid AH (2022) Oxidative stress-induced endothelial dysfunction in cardiovascular diseases. Front Biosci (landmark Ed) 27:105. https://doi.org/10.31083/j.fbl2703105
Shan D, Qu P, Zhong C, He L, Zhang Q, Zhong G, Hu W, Feng Y, Yang S, Yang XF, Yu J (2022) Anemoside B4 inhibits vascular smooth muscle cell proliferation, migration, and neointimal hyperplasia. Front Cardiovasc Med 9:907490. https://doi.org/10.3389/fcvm.2022.907490
Shlofmitz E, Iantorno M, Waksman R (2019) Restenosis of drug-eluting stents: a new classification system based on disease mechanism to guide treatment and state-of-the-art review. Circ Cardiovasc Interv 12:e007023. https://doi.org/10.1161/CIRCINTERVENTIONS.118.007023
Siontis GC, Stefanini GG, Mavridis D, Siontis KC, Alfonso F, Perez-Vizcayno MJ, Byrne RA, Kastrati A, Meier B, Salanti G, Juni P, Windecker S (2015) Percutaneous coronary interventional strategies for treatment of in-stent restenosis: a network meta-analysis. Lancet 386:655–664. https://doi.org/10.1016/S0140-6736(15)60657-2
Song JB, Shen J, Fan J, Zhang Z, Yi ZJ, Bai S, Mu XL, Xiao L (2020) Effects of a matrix metalloproteinase inhibitor-eluting stent on in-stent restenosis. Med Sci Monit 26:e922556. https://doi.org/10.12659/MSM.922556
Tang HY, Chen AQ, Zhang H, Gao XF, Kong XQ, Zhang JJ (2022) Vascular smooth muscle cells phenotypic switching in cardiovascular diseases. Cells. https://doi.org/10.3390/cells11244060
Todd DJ, Lee AH, Glimcher LH (2008) The endoplasmic reticulum stress response in immunity and autoimmunity. Nat Rev Immunol 8:663–674. https://doi.org/10.1038/nri2359
Wang S, Zhang M, Liu Z, Yang W, Shi J, Dean V, Chen D (2017) Relationship between CHOP/GADD153 and unstable human carotid atherosclerotic plaque. Br J Neurosurg 31:648–652. https://doi.org/10.1080/02688697.2017.1327016
Wang X, Wei W, Wu J, Kang L, Wu S, Li J, Shen Y, You J, Ye Y, Zhang Q, Zou Y (2022) Involvement of endoplasmic reticulum stress-mediated activation of C/EBP homologous protein in aortic regurgitation-induced cardiac remodeling in mice. J Cardiovasc Transl Res 15:340–349. https://doi.org/10.1007/s12265-021-10162-4
Wininger KL (2022) Percutaneous transluminal coronary angioplasty: history, current techniques, and future directions. Radiol Technol 94:35–45
Yang Y, Liu L, Naik I, Braunstein Z, Zhong J, Ren B (2017) Transcription factor C/EBP homologous protein in health and diseases. Front Immunol 8:1612. https://doi.org/10.3389/fimmu.2017.01612
Yang X, Yang Y, Guo J, Meng Y, Li M, Yang P, Liu X, Aung LHH, Yu T, Li Y (2021) Targeting the epigenome in in-stent restenosis: from mechanisms to therapy. Mol Ther Nucleic Acids 23:1136–1160. https://doi.org/10.1016/j.omtn.2021.01.024
Yoshida T, Sugiura H, Mitobe M, Tsuchiya K, Shirota S, Nishimura S, Shiohira S, Ito H, Nobori K, Gullans SR, Akiba T, Nitta K (2008) ATF3 protects against renal ischemia-reperfusion injury. J Am Soc Nephrol 19:217–224. https://doi.org/10.1681/ASN.2005111155
Zhang DM, Chen SL (2021) Potential Mechanisms of In-stent neointimal atherosclerotic plaque formation. J Cardiovasc Pharmacol 78:388–393. https://doi.org/10.1097/FJC.0000000000001059
Zhou X, Lu B, Fu D, Gui M, Yao L, Li J (2020) Huoxue Qianyang decoction ameliorates cardiac remodeling in obese spontaneously hypertensive rats in association with ATF6-CHOP endoplasmic reticulum stress signaling pathway regulation. Biomed Pharmacother 121:109518. https://doi.org/10.1016/j.biopha.2019.109518
Zhou Z, Fan Y, Zong R, Tan K (2022) The mitochondrial unfolded protein response: a multitasking giant in the fight against human diseases. Ageing Res Rev 81:101702. https://doi.org/10.1016/j.arr.2022.101702
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This study was supported by grants from the Hubei Provincial Department of Education Science Foundation of China (Grant No. B2022036).
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LT and JY designed the study and wrote the original draft; QQ, Z-yZ, FZ and C-yC completed experiments and CH analyzed data; J-wD and JY revised the manuscript. The final version was reviewed by all authors.
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Teng, L., Qin, Q., Zhou, Zy. et al. Role of C/EBP Homologous Protein in Vascular Stenosis After Carotid Artery Injury. Biochem Genet (2024). https://doi.org/10.1007/s10528-024-10713-9
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DOI: https://doi.org/10.1007/s10528-024-10713-9