Thrombospondin‑2 is upregulated in patients with aortic dissection and enhances angiotensin II‑induced smooth muscle cell apoptosis

Qi,Liping; Wu,Kui; Shi,Shutian; Ji,Qingwei; Miao,Huangtai; Bin,Que

doi:10.3892/etm.2020.9279

Thrombospondin‑2 is upregulated in patients with aortic dissection and enhances angiotensin II‑induced smooth muscle cell apoptosis

Authors:
- Liping Qi
- Kui Wu
- Shutian Shi
- Qingwei Ji
- Huangtai Miao
- Que Bin
View Affiliations

Affiliations: Department of Cardiology, The Second Clinical Center, Chinese People's Liberation Army General Hospital, Beijing 100853, P.R. China, Emergency and Critical Care Center, Beijing Anzhen Hospital, Capital Medical University, Beijing Institute of Heart, Lung, and Blood Vessel Diseases, Beijing Lab for Cardiovascular Precision Medicine, Beijing 100029, P.R. China
Published online on: October 6, 2020 https://doi.org/10.3892/etm.2020.9279
Article Number: 150
Copyright: © Qi 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

Thrombospondin‑2 (TSP‑2) is an important extracellular matrix protein that is involved in a variety of cardiovascular diseases, including viral myocarditis and abdominal aortic aneurysm. The present study aimed to investigate TSP‑2 expression in patients with aortic dissection (AD). Aortas were obtained from patients with AD and healthy donors, and TSP‑2 expression level in all samples was measured by western blotting and immunofluorescence assays. Blood samples were also collected from patients with AD and non‑AD (NAD) subjects. Circulating TSP‑2, tumor necrosis factor (TNF)‑α and interleukin (IL)‑6 levels in each sample were detected using ELISAs. In addition, the effect of TSP‑2 on angiotensin II (Ang II)‑induced smooth muscle cell (SMC) apoptosis was assessed in vitro. Compared with healthy donors, aortic TSP‑2 expression level was significantly increased in patients with AD. Furthermore, TSP‑2 was secreted primarily by SMCs, but also by endothelial cells. TSP‑2 plasma expression level was also elevated in patients with AD compared with non‑AD subjects. Furthermore, TSP‑2 serum expression level was positively correlated with TNF‑α and IL‑6 expression levels in patients with AD. In addition, recombinant mouse TSP‑2 treatment increased Bax mRNA expression and decreased Bcl2 mRNA expression in Ang II‑treated SMCs; however, the effects were reversed following treatment with the NF‑κB p65 signaling pathway inhibitor JSH‑23 or with the anti‑TNF‑α and anti‑IL‑6 neutralizing antibodies. The present study demonstrated that TSP‑2 expression was increased in the aortic tissues and plasma of patients with AD. These findings suggested that TSP‑2 may participate in the progression of AD by activating the NF‑κB p65 signaling pathway and amplifying the inflammatory response.

View Figures

View References

1	Hagan PG, Nienaber CA, Isselbacher EM, Bruckman D, Karavite DJ, Russman PL, Evangelista A, Fattori R, Suzuki T, Oh JK, et al: The international registry of acute aortic dissection (IRAD): New insights into an old disease. JAMA. 283:897–903. 2000.PubMed/NCBI View Article : Google Scholar
2	Chen LW, Wu XJ, Lu L, Zhang GC, Yang GF, Yang ZW, Dong Y, Cao H and Chen Q: Total arch repair for acute type A aortic dissection with 2 modified techniques: Open single-branched stent graft placement and reinforcement of the dissected arch vessel stump with stent graft. Circulation. 123:2536–2541. 2011.PubMed/NCBI View Article : Google Scholar
3	Fattori R, Cao P, De Rango P, Czerny M, Evangelista A, Nienaber C, Rousseau H and Schepens M: Interdisciplinary expert consensus document on management of type B aortic dissection. J Am Coll Cardiol. 61:1661–1678. 2013.PubMed/NCBI View Article : Google Scholar
4	Hugo C and Daniel C: Thrombospondin in renal disease. Nephron Exp Nephrol. 111:e61–e66. 2009.PubMed/NCBI View Article : Google Scholar
5	Calabro NE, Kristofik NJ and Kyriakides TR: Thrombospondin-2 and extracellular matrix assembly. Biochim Biophys Acta. 1840:2396–2402. 2014.PubMed/NCBI View Article : Google Scholar
6	Rusnati M, Borsotti P, Moroni E, Foglieni C, Chiodelli P, Carminati L, Pinessi D, Annis DS, Paiardi G, Bugatti A, et al: The calcium-binding type III repeats domain ofthrombospondin-2 binds to fibroblast growth factor 2 (FGF2). Angiogenesis. 22:133–144. 2019.PubMed/NCBI View Article : Google Scholar
7	Morris AH, Stamer DK, Kunkemoeller B, Chang J, Xing H and Kyriakides TR: Decellularized materials derived from TSP2-KO mice promote enhanced neovascularization and integration in diabetic wounds. Biomaterials. 169:61–71. 2018.PubMed/NCBI View Article : Google Scholar
8	Helkin A, Maier KG and Gahtan V: Thrombospondin-1, -2 and -5 have differential effects on vascular smooth muscle cell physiology. Biochem Biophys Res Commun. 464:1022–1027. 2015.PubMed/NCBI View Article : Google Scholar
9	Daniel C, Vogelbacher R, Stief A, Grigo C and Hugo C: Long-term gene therapy with thrombospondin 2 inhibits TGF-β activation, inflammation and angiogenesis in chronic allograft nephropathy. PLoS One. 8(e83846)2013.PubMed/NCBI View Article : Google Scholar
10	Papageorgiou AP, Swinnen M, Vanhoutte D, VandenDriessche T, Chuah M, Lindner D, Verhesen W, de Vries B, D'hooge J, Lutgens E, et al: Thrombospondin-2 prevents cardiac injury and dysfunction in viral myocarditis through the activation of regulatory T-cells. Cardiovasc Res. 94:115–124. 2012.PubMed/NCBI View Article : Google Scholar
11	Li K, Huang EP, Su J, Zhu P, Lin J, Luo SQ and Yang CL: Therapeutic role for TSP-2 antibody in a murine asthma model. Int Arch Allergy Immunol. 175:160–170. 2018.PubMed/NCBI View Article : Google Scholar
12	Kimura Y, Izumiya Y, Hanatani S, Yamamoto E, Kusaka H, Tokitsu T, Takashio S, Sakamoto K, Tsujita K, Tanaka T, et al: High serum levels of thrombospondin-2 correlate with poor prognosis of patients with heart failure with preserved ejection fraction. Heart Vessels. 31:52–59. 2016.PubMed/NCBI View Article : Google Scholar
13	Hanatani S, Izumiya Y, Takashio S, Kimura Y, Araki S, Rokutanda T, Tsujita K, Yamamoto E, Tanaka T, Yamamuro M, et al: Circulating thrombospondin-2 reflects disease severity and predicts outcome of heart failure with reduced ejection fraction. Circ J. 78:903–910. 2014.PubMed/NCBI View Article : Google Scholar
14	Golledge J, Clancy P, Hankey GJ and Norman PE: Relation between serum thrombospondin-2 and cardiovascular mortality in older men screened for abdominal aortic aneurysm aneurysm. Am J Cardiol. 111:1800–1804. 2013.PubMed/NCBI View Article : Google Scholar
15	van Almen GC, Swinnen M, Carai P, Verhesen W, Cleutjens JP, D'hooge J, Verheyen FK, Pinto YM, Schroen B, Carmeliet P and Heymans S: Absence of thrombospondin-2 increases cardiomyocyte damage and matrix disruption in doxorubicin-induced cardiomyopathy. J Mol Cell Cardiol. 51:318–328. 2011.PubMed/NCBI View Article : Google Scholar
16	Swinnen M, Vanhoutte D, Van Almen GC, Hamdani N, Schellings MW, D'hooge J, Van der Velden J, Weaver MS, Sage EH, Bornstein P, et al: Absence of thrombospondin-2 causes age-related dilated cardiomyopathy. Circulation. 120:1585–1597. 2009.PubMed/NCBI View Article : Google Scholar
17	Xiao L, Kirabo A, Wu J, Saleh MA, Zhu L, Wang F, Takahashi T, Loperena R, Foss JD, Mernaugh RL, et al: Renal denervation prevents immune cell activation and renal inflammation in angiotensin II-induced hypertension. Circ Res. 117:547–557. 2015.PubMed/NCBI View Article : Google Scholar
18	Li Y, Zhang C, Wu Y, Han Y, Cui W, Jia L, Cai L, Cheng J, Li H and Du J: Interleukin-12p35 deletion promotes CD4 T-cell-dependent macrophage differentiation and enhances angiotensin II-Induced cardiac fibrosis. Arterioscler Thromb Vasc Biol. 32:1662–1674. 2012.PubMed/NCBI View Article : Google Scholar
19	Lake FR, Noble PW, Henson PM and Riches DW: Functional switching of macrophage responses to tumor necrosis factor-alpha (TNF alpha) by interferons. Implications for the pleiotropic activities of TNF alpha. J Clin Invest. 93:1661–1669. 1994.PubMed/NCBI View Article : Google Scholar
20	Golovina VA and Blaustein MP: Preparation of primary cultured mesenteric artery smooth muscle cells for fluorescent imaging and physiological studies. Nat Protoc. 1:2681–2687. 2006.PubMed/NCBI View Article : Google Scholar
21	Jia LX, Zhang WM, Zhang HJ, Li TT, Wang YL, Qin YW, Gu H and Du J: Mechanical stretch-induced endoplasmic reticulum stress, apoptosis and inflammation contribute to thoracic aortic aneurysm and dissection. J Pathol. 236:373–383. 2015.PubMed/NCBI View Article : Google Scholar
22	Kobayashi M, Inoue K, Warabi E, Minami T and Kodama T: A simple method of isolating mouse aortic endothelial cells. J Atheroscler Thromb. 12:138–142. 2005.PubMed/NCBI View Article : Google Scholar
23	Ye J, Que B, Huang Y, Lin Y, Chen J, Liu L, Shi Y, Wang Y, Wang M, Zeng T, et al: Interleukin-12p35 knockout promotes macrophage differentiation, aggravates vascular dysfunction, and elevates blood pressure in angiotensin II-infused mice. Cardiovasc Res. 115:1102–1113. 2019.PubMed/NCBI View Article : Google Scholar
24	Bae ON, Wang JM, Baek SH, Wang Q, Yuan H and Chen AF: Oxidative stress-mediated thrombospondin-2 upregulation impairs bone marrow-derived angiogenic cell function in diabetes mellitus. Arterioscler Thromb Vasc Biol. 33:1920–1927. 2013.PubMed/NCBI View Article : Google Scholar
25	Xiao T, Zhang L, Huang Y, Shi Y, Wang J, Ji Q, Ye J, Lin Y and Liu H: Sestrin2 increases in aortas and plasma from aortic dissection patients and alleviates angiotensin II-induced smooth muscle cell apoptosis via the Nrf2 pathway. Life Sci. 218:132–138. 2019.PubMed/NCBI View Article : Google Scholar
26	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.PubMed/NCBI View Article : Google Scholar
27	Pohjolainen V, Mustonen E, Taskinen P, Näpänkangas J, Leskinen H, Ohukainen P, Peltonen T, Aro J, Juvonen T, Satta J, et al: Increased thrombospondin-2 in human fibrosclerotic and stenotic aortic valves. Atherosclerosis. 220:66–71. 2012.PubMed/NCBI View Article : Google Scholar
28	Ortega R, Collado A, Selles F, Gonzalez-Navarro H, Sanz MJ, Real JT and Piqueras J: SGLT-2 (Sodium-Glucose Cotransporter 2) inhibition reduces Ang II (Angiotensin II)-induced dissecting abdominal aortic aneurysm in ApoE (Apolipoprotein E) knockout mice. Arterioscler Thromb Vasc Biol. 39:1614–1628. 2019.PubMed/NCBI View Article : Google Scholar
29	Ye J, Wang Y, Wang Z, Ji Q, Huang Y, Zeng T, Hu H, Ye D, Wan J and Lin Y: Circulating Th1, Th2, Th9, Th17, Th22, and treg levels in aortic dissection patients. Mediators Inflamm. 2018(5697149)2018.PubMed/NCBI View Article : Google Scholar
30	Zeng T, Shi L, Ji Q, Shi Y, Huang Y, Liu Y, Gan J, Yuan J, Lu Z, Xue Y, et al: Cytokines in aortic dissection. Clin Chim Acta. 486:177–182. 2018.PubMed/NCBI View Article : Google Scholar
31	El-Hamamsy I and Yacoub MH: Cellular and molecular mechanisms of thoracic aortic aneurysms. Nat Rev Cardiol. 6:771–786. 2009.PubMed/NCBI View Article : Google Scholar
32	Suzuki T, Katoh H, Watanabe M, Kurabayashi M, Hiramori K, Hori S, Nobuyoshi M, Tanaka H, Kodama K, Sato H, et al: Novel biochemical diagnostic method for aortic dissection. Results of a prospective study using an immunoassay of smooth muscle myosin heavy chain. Circulation. 93:1244–1249. 1996.PubMed/NCBI View Article : Google Scholar
33	Michel JB, Jondeau G and Milewicz DM: From genetics to response to injury: Vascular smooth muscle cells in aneurysms and dissections of the ascending aorta. Cardiovasc Res. 114:578–589. 2018.PubMed/NCBI View Article : Google Scholar
34	Ye P, Chen W, Wu J, Huang X, Li J, Wang S, Liu Z, Wang G, Yang X, Zhang P, et al: GM-CSF contributes to aortic aneurysms resulting from SMAD3 deficiency. J Clin Invest. 123:2317–2331. 2013.PubMed/NCBI View Article : Google Scholar
35	De Stefano D, Nicolaus G, Maiuri MC, Cipolletta D, Galluzzi L, Cinelli MP, Tajana G, Iuvone T and Carnuccio R: NF-kappaB blockade upregulates Bax, TSP-1, and TSP-2 expression in rat granulation tissue. J Mol Med (Berl). 87:481–492. 2009.PubMed/NCBI View Article : Google Scholar