LincRNA‑EPS increases TGF‑β expression to inhibit the Wnt/β‑catenin pathway, VSMC osteoblastic differentiation and vascular calcification in diabetic mice

Li,Yibo; Xi,Ziwei; Yu,Zheng; Yang,Chaoyue; Tan,Chunhua

doi:10.3892/etm.2022.11352

LincRNA‑EPS increases TGF‑β expression to inhibit the Wnt/β‑catenin pathway, VSMC osteoblastic differentiation and vascular calcification in diabetic mice

Authors:
- Yibo Li
- Ziwei Xi
- Zheng Yu
- Chaoyue Yang
- Chunhua Tan
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Affiliations: Department of Orthopedics, General Hospital of Central Theater Command (Hankou District), Wuhan, Hubei 430000, P.R. China, School of Medical Sciences, Army Medical University, Chongqing 400038, P.R. China, Department of Hematoendocrinology, 32295 Army Hospital, Liaoyang, Liaoning 111000, P.R. China
Published online on: May 4, 2022 https://doi.org/10.3892/etm.2022.11352
Article Number: 425
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

In patients with diabetes, the Wnt/β‑catenin pathway in vascular smooth muscle cells (VSMCs) is continuously activated by low‑intensity inflammation, which leads to the osteoblastic differentiation of these cells and the deposition of calcium and phosphorus in blood vessels. The aim of the present study was to determine whether long intergenic non‑coding RNA‑erythroid pro‑survival (lincRNA‑EPS) was able to ameliorate vascular calcification (VC) associated with diabetes. VSMCs isolated from C57BL/6 mice were transfected with lincRNA‑EPS overexpression vector in vitro and their osteoblastic differentiation was evaluated under high‑glucose conditions. In addition, a mouse model of diabetes was established, which included a lincRNA‑EPS knockout group and a lincRNA‑EPS high expression group. Blood vessel samples from the mice were examined to determine the degree of calcification. The levels of inflammatory factors in serum were also detected. The VSMCs transfected with lincRNA‑EPS overexpression vector exhibited less osteoblastic differentiation and migration and significantly lower levels of Wnt pathway‑associated proteins than those transfected with empty control. Furthermore, the in vivo experiments revealed that the overexpression of lincRNA‑EPS significantly reduced VC in diabetic mice. Therefore, on the basis of these findings, it is suggested that lincRNA‑EPS overexpression may provide a novel and effective method for the treatment of VC in patients with diabetes.

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1	American Diabetes Association. Diagnosis and classification of diabetes mellitus. Diabetes Care. 36 (Suppl 1):S67–S74. 2013.PubMed/NCBI View Article : Google Scholar
2	Zhang C, Zhang K, Huang F, Feng W, Chen J, Zhang H, Wang J, Luo P and Huang H: Exosomes, the message transporters in vascular calcification. J Cell Mol Med. 22:4024–4033. 2018.PubMed/NCBI View Article : Google Scholar
3	Neuenschwander M, Ballon A, Weber KS, Norat T, Aune D, Schwingshackl L and Schlesinger S: Role of diet in type 2 diabetes incidence: Umbrella review of meta-analyses of prospective observational studies. BMJ. 366(l2368)2019.PubMed/NCBI View Article : Google Scholar
4	Li XY, Li QM, Fang Q, Zha XQ, Pan LH and Luo JP: Laminaria japonica polysaccharide inhibits vascular calcification via preventing osteoblastic differentiation of vascular smooth muscle cells. J Agric Food Chem. 66:1821–1827. 2018.PubMed/NCBI View Article : Google Scholar
5	Zhan JK, Wang YJ, Wang Y, Tang ZY, Tan P, Huang W and Liu YS: The protective effect of GLP-1 analogue in arterial calcification through attenuating osteoblastic differentiation of human VSMCs. Int J Cardiol. 189:188–193. 2015.PubMed/NCBI View Article : Google Scholar
6	Pedersen BK: Anti-inflammatory effects of exercise: Role in diabetes and cardiovascular disease. Eur J Clin Invest. 47:600–611. 2017.PubMed/NCBI View Article : Google Scholar
7	Nagano A, Arioka M, Takahashi-Yanaga F, Matsuzaki E and Sasaguri T: Celecoxib inhibits osteoblast maturation by suppressing the expression of Wnt target genes. J Pharmacol Sci. 133:18–24. 2017.PubMed/NCBI View Article : Google Scholar
8	Clevers H and Nusse R: Wnt/β-catenin signaling and disease. Cell. 149:1192–1205. 2012.PubMed/NCBI View Article : Google Scholar
9	Ravi R, Noonan KA, Pham V, Bedi R, Zhavoronkov A, Ozerov IV, Makarev E, V Artemov A, Wysocki PT, Mehra R, et al: Bifunctional immune checkpoint-targeted antibody-ligand traps that simultaneously disable TGFβ enhance the efficacy of cancer immunotherapy. Nat Commun. 9(741)2018.PubMed/NCBI View Article : Google Scholar
10	Bierie B and Moses HL: Transforming growth factor beta (TGF-beta) and inflammation in cancer. Cytokine Growth Factor Rev. 21:49–59. 2010.PubMed/NCBI View Article : Google Scholar
11	Atianand MK, Hu W, Satpathy AT, Shen Y, Ricci EP, Alvarez-Dominguez JR, Bhatta A, Schattgen SA, McGowan JD, Blin J, et al: A long noncoding RNA lincRNA-EPS acts as a transcriptional brake to restrain inflammation. Cell. 165:1672–1685. 2016.PubMed/NCBI View Article : Google Scholar
12	Ke Z, Lu J, Zhu J, Yang Z, Jin Z and Yuan L: Down-regulation of lincRNA-EPS regulates apoptosis and autophagy in BCG-infected RAW264.7 macrophages via JNK/MAPK signaling pathway. Infect Genet Evol. 77(104077)2020.PubMed/NCBI View Article : Google Scholar
13	Heydemann A: An overview of murine high fat diet as a model for type 2 diabetes mellitus. J Diabetes Res. 2016(2902351)2016.PubMed/NCBI View Article : Google Scholar
14	Azushima K, Gurley SB and Coffman TM: Modelling diabetic nephropathy in mice. Nat Rev Nephrol. 14:48–56. 2018.PubMed/NCBI View Article : Google Scholar
15	Schwarz H, Zhang Y, Zhan C, Malm M, Field R, Turner R, Sellick C, Varley P, Rockberg J and Chotteau V: Small-scale bioreactor supports high density HEK293 cell perfusion culture for the production of recombinant Erythropoietin. J Biotechnol. 309:44–52. 2020.PubMed/NCBI View Article : Google Scholar
16	Sun X, Li W, Zhang X, Qi M, Zhang Z, Zhang XE and Cui Z: In vivo targeting and imaging of atherosclerosis using multifunctional virus-like particles of simian virus 40. Nano Lett. 16:6164–6171. 2016.PubMed/NCBI View Article : Google Scholar
17	Grieger JC, Soltys SM and Samulski RJ: Production of recombinant adeno-associated virus vectors using suspension HEK293 cells and continuous harvest of vector from the culture media for GMP FIX and FLT1 clinical vector. Mol Ther. 24:287–297. 2016.PubMed/NCBI View Article : Google Scholar
18	Chandler RJ, LaFave MC, Varshney GK, Trivedi NS, Carrillo-Carrasco N, Senac JS, Wu W, Hoffmann V, Elkahloun AG, Burgess SM and Venditti CP: Vector design influences hepatic genotoxicity after adeno-associated virus gene therapy. J Clin Invest. 125:870–880. 2015.PubMed/NCBI View Article : Google Scholar
19	Wei J, Ran G, Wang X, Jiang N, Liang J, Lin X, Ling C and Zhao B: Gene manipulation in liver ductal organoids by optimized recombinant adeno-associated virus vectors. J Biol Chem. 294:14096–14104. 2019.PubMed/NCBI View Article : Google Scholar
20	Li Y, Wan S, Liu G, Cai W, Huo D, Li G, Yang M, Wang Y, Guan G, Ding N, et al: Netrin-1 promotes inflammation resolution to achieve endothelialization of small-diameter tissue engineering blood vessels by improving endothelial progenitor cells function in situ. Adv Sci (Weinh). 4(1700278)2017.PubMed/NCBI View Article : Google Scholar
21	Jiao L, Zhuang Y, Jiang M, Zhou JH, Wu M, Chen ZJ, Fang JH and Deng YS: Angiopoietin-like 2 has auxo-action in atherosclerosis by promoting atherosclerotic calcification. Int J Clin Exp Pathol. 10:9084–9091. 2017.PubMed/NCBI
22	Fang M, Wang CG, Zheng C, Luo J, Hou S, Liu K and Li X: Mir-29b promotes human aortic valve interstitial cell calcification via inhibiting TGF-β3 through activation of wnt3/β-catenin/Smad3 signaling. J Cell Biochem. 119:5175–5185. 2018.PubMed/NCBI View Article : Google Scholar
23	Zhang B, Li Q, Jia S, Li F, Li Q and Li J: LincRNA-EPS in biomimetic vesicles targeting cerebral infarction promotes inflammatory resolution and neurogenesis. J Transl Med. 18(110)2020.PubMed/NCBI View Article : Google Scholar
24	Zou Y, Zhang Y, Church J and Liu X: Comparison of β-catenin and LEF1 immunohistochemical stains in desmoid-type fibromatosis and its selected mimickers, with unexpected finding of LEF1 positivity in scars. Appl Immunohistochem Mol Morphol. 26:648–653. 2018.PubMed/NCBI View Article : Google Scholar
25	Wang Q, Wang G, Wang B and Yang H: Activation of TGR5 promotes osteoblastic cell differentiation and mineralization. Biomed Pharmacother. 108:1797–1803. 2018.PubMed/NCBI View Article : Google Scholar
26	He J, Zhang N, Zhang J, Jiang B and Wu F: Migration critically meditates osteoblastic differentiation of bone mesenchymal stem cells through activating canonical Wnt signal pathway. Colloids Surf B Biointerfaces. 171:205–213. 2018.PubMed/NCBI View Article : Google Scholar
27	Shioi A and Ikari Y: Plaque calcification during atherosclerosis progression and regression. J Atheroscler Thromb. 25:294–303. 2018.PubMed/NCBI View Article : Google Scholar
28	Chiarella E, Aloisio A, Scicchitano S, Lucchino V, Montalcini Y, Galasso O, Greco M, Gasparini G, Mesuraca M, Bond HM and Morrone G: ZNF521 represses osteoblastic differentiation in human adipose-derived stem cells. Int J Mol Sci. 19(4095)2018.PubMed/NCBI View Article : Google Scholar
29	Liu Y, Lin F, Fu Y, Chen W, Liu W, Chi J, Zhang X and Yin X: Cortistatin inhibits calcification of vascular smooth muscle cells by depressing osteoblastic differentiation and endoplasmic reticulum stress. Amino Acids. 48:2671–2681. 2016.PubMed/NCBI View Article : Google Scholar
30	Rong S, Zhao X, Jin X, Zhang Z, Chen L, Zhu Y and Yuan W: Vascular calcification in chronic kidney disease is induced by bone morphogenetic protein-2 via a mechanism involving the Wnt/β-catenin pathway. Cell Physiol Biochem. 34:2049–2060. 2014.PubMed/NCBI View Article : Google Scholar
31	Nagashima M, Sakai A, Uchida S, Tanaka S, Tanaka M and Nakamura T: Bisphosphonate (YM529) delays the repair of cortical bone defect after drill-hole injury by reducing terminal differentiation of osteoblasts in the mouse femur. Bone. 36:502–511. 2005.PubMed/NCBI View Article : Google Scholar
32	Rudijanto A: The role of vascular smooth muscle cells on the pathogenesis of atherosclerosis. Acta Med Indones. 39:86–93. 2007.PubMed/NCBI
33	Guzik TJ, Korbut R and Adamek-Guzik T: Nitric oxide and superoxide in inflammation and immune regulation. J Physiol Pharmacol. 54:469–487. 2003.PubMed/NCBI
34	Beck PL, Xavier R, Wong J, Ezedi I, Mashimo H, Mizoguchi A, Mizoguchi E, Bhan AK and Podolsky DK: Paradoxical roles of different nitric oxide synthase isoforms in colonic injury. Am J Physiol Gastrointest Liver Physiol. 286:G137–G147. 2004.PubMed/NCBI View Article : Google Scholar
35	Niedbala W, Besnard AG, Nascimento DC, Donate PB, Sonego F, Yip E, Guabiraba R, Chang HD, Fukada SY, Salmond RJ, et al: Nitric oxide enhances Th9 cell differentiation and airway inflammation. Nat Commun. 5(4575)2014.PubMed/NCBI View Article : Google Scholar
36	de Seigneux S and Martin PY: Phosphate and FGF23 in the renoprotective benefit of RAAS inhibition. Pharmacol Res. 106:87–91. 2016.PubMed/NCBI View Article : Google Scholar
37	Jüppner H: Phosphate and FGF-23. Kidney Int Suppl. 79:S24–S27. 2011.PubMed/NCBI View Article : Google Scholar