Overexpression of microRNA‑155 alleviates palmitate‑induced vascular endothelial cell injury in human umbilical vein endothelial cells by negatively regulating the Wnt signaling pathway

Zhao,Yanping; Rao,Wenquan; Wan,Yanna; Yang,Xinglong; Wang,Guorong; Deng,Jun; Dai,Min; Liu,Qiang

doi:10.3892/mmr.2019.10623

Overexpression of microRNA‑155 alleviates palmitate‑induced vascular endothelial cell injury in human umbilical vein endothelial cells by negatively regulating the Wnt signaling pathway

Authors:
- Yanping Zhao
- Wenquan Rao
- Yanna Wan
- Xinglong Yang
- Guorong Wang
- Jun Deng
- Min Dai
- Qiang Liu
View Affiliations

Affiliations: Department of Vascular Surgery, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China, Department of Urology Surgery, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China
Published online on: August 27, 2019 https://doi.org/10.3892/mmr.2019.10623
Pages: 3527-3534
Copyright: © Zhao 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

The present study aimed to investigate the effect of microRNA155 (miR‑155) on palmitate‑induced vascular endothelial cell injury in human umbilical vein endothelial cells (HUVECs) via the regulation of the Wnt signaling pathway. HUVECs were treated with 0.1 mM palmitate. After transfection with mimic, antagomir or the Wnt pathway inhibitor XAV939, HUVECs were divided into six treatment groups: Control, palmitate, mimic + palmitate, mimic + palmitate + XAV939, antagomir + palmitate, antagomir + palmitate + XAV939. miR‑155 expression was detected using reverse transcription‑quantitative PCR. The expression levels of the Wnt signaling pathway‑related factors β‑catenin and Cyclin D, and the inflammatory factors interleukin‑6 (IL‑6) and tumor necrosis factor‑α (TNF‑α), were detected using western blot analysis. MTT and Transwell assays were used to detect the proliferation and migration of cells, respectively. Apoptosis and reactive oxygen species (ROS) levels were determined using flow cytometry. The localization of β‑catenin in cells was determined by immunofluorescence. Palmitate reduced the expression level of miR‑155 in HUVECs. In palmitate‑induced HUVECs, overexpression of miR‑155 promoted cell proliferation, reduced the levels of apoptosis, downregulated IL‑6 and TNF‑α expression, and reduced ROS levels. Inhibition of the Wnt signaling pathway enhanced the anti‑endothelial cell injury effect caused by the overexpression of miR‑155 in palmitate‑induced HUVECs, thereby promoting proliferation, reducing apoptosis, downregulating the levels of inflammatory factors and reducing ROS levels. In summary, overexpression of miR‑155 inhibited palmitate‑induced apoptosis, ROS production and levels of inflammatory factors, and promoted the proliferation of HUVECs by negatively regulating the Wnt signaling pathway. This present study provides a theoretical basis for the prevention and treatment of cardiovascular diseases associated with endothelial cell injury.

View Figures

View References

1	Fleissner F and Thum T: Critical role of the nitric oxide/reactive oxygen species balance in endothelial progenitor dysfunction. Antioxid Redox Signal. 15:933–948. 2011. View Article : Google Scholar : PubMed/NCBI
2	Bollu LR, Katreddy RR, Blessing AM, Pham N, Zheng B, Wu X and Weihua Z: Intracellular activation of EGFR by fatty acid synthase dependent palmitoylation. Oncotarget. 6:34992–35003. 2015. View Article : Google Scholar : PubMed/NCBI
3	Zhu H, Yang Y, Wang Y, Li J, Schiller PW and Peng T: MicroRNA-195 promotes palmitate-induced apoptosis in cardiomyocytes by down-regulating Sirt1. Cardiovasc Res. 92:75–84. 2011. View Article : Google Scholar : PubMed/NCBI
4	Gao X, Zhao XL, Zhu YH, Li XM, Xu Q, Lin HD and Wang MW: Tetramethylpyrazine protects palmitate-induced oxidative damage and mitochondrial dysfunction in C2C12 myotubes. Life Sci. 88:803–809. 2011. View Article : Google Scholar : PubMed/NCBI
5	Liu K, Zhao W, Gao X, Huang F, Kou J and Liu B: Diosgenin ameliorates palmitate-induced endothelial dysfunction and insulin resistance via blocking IKKβ and IRS-1 pathways. Atherosclerosis. 223:350–358. 2012. View Article : Google Scholar : PubMed/NCBI
6	Ogawa Y, Imajo K, Honda Y, Kessoku T, Tomeno W, Kato S, Fujita K, Yoneda M, Saito S, Saigusa Y, et al: Palmitate-induced lipotoxicity is crucial for the pathogenesis of nonalcoholic fatty liver disease in cooperation with gut-derived endotoxin. Sci Rep. 8:113652018. View Article : Google Scholar : PubMed/NCBI
7	Zhang M, Wang CM, Li J, Meng ZJ, Wei SN, Li J, Bucala R, Li YL and Chen L: Berberine protects against palmitate-induced endothelial dysfunction: Involvements of upregulation of AMPK and eNOS and downregulation of NOX4. Mediators Inflamm. 2013:2604642013. View Article : Google Scholar : PubMed/NCBI
8	Artwohl M, Roden M, Waldhäusl W, Freudenthaler A and Baumgartner-Parzer SM: Free fatty acids trigger apoptosis and inhibit cell cycle progression in human vascular endothelial cells. FASEB J. 18:146–148. 2004. View Article : Google Scholar : PubMed/NCBI
9	Sonkoly E, Janson P, Majuri ML, Savinko T, Fyhrquist N, Eidsmo L, Xu N, Meisgen F, Wei T, Bradley M, et al: MiR-155 is overexpressed in patients with atopic dermatitis and modulates T-cell proliferative responses by targeting cytotoxic T lymphocyte-associated antigen 4. J Allergy Clin Immunol. 126:581–589 e1-e20. 2010. View Article : Google Scholar : PubMed/NCBI
10	Yang Q, Zhang Q, Qing Y, Zhou L, Mi Q and Zhou J: miR-155 is dispensable in monosodium urate-induced gouty inflammation in mice. Arthritis Res Ther. 20:1442018. View Article : Google Scholar : PubMed/NCBI
11	Babar IA, Cheng CJ, Booth CJ, Liang X, Weidhaas JB, Saltzman WM and Slack FJ: Nanoparticle-based therapy in an in vivo microRNA-155 (miR-155)-dependent mouse model of lymphoma. Proc Natl Acad Sci USA. 109:E1695–E1704. 2012. View Article : Google Scholar : PubMed/NCBI
12	Pulkkinen KH, Yla-Herttuala S and Levonen AL: Heme oxygenase 1 is induced by miR-155 via reduced BACH1 translation in endothelial cells. Free Radic Biol Med. 51:2124–2131. 2011. View Article : Google Scholar : PubMed/NCBI
13	Clevers H and Nusse R: Wnt/β-catenin signaling and disease. Cell. 149:1192–1205. 2012. View Article : Google Scholar : PubMed/NCBI
14	Marinou K, Christodoulides C, Antoniades C and Koutsilieris M: Wnt signaling in cardiovascular physiology. Trends Endocrinol Metab. 23:628–636. 2012. View Article : Google Scholar : PubMed/NCBI
15	Dawson K, Aflaki M and Nattel S: Role of the Wnt-Frizzled system in cardiac pathophysiology: A rapidly developing, poorly understood area with enormous potential. J Physiol. 591:1409–1432. 2013. View Article : Google Scholar : PubMed/NCBI
16	Ma S, Yao S, Tian H, Jiao P, Yang N, Zhu P and Qin S: Pigment epithelium-derived factor alleviates endothelial injury by inhibiting Wnt/β-catenin pathway. Lipids Health Dis. 16:312017. View Article : Google Scholar : PubMed/NCBI
17	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 : PubMed/NCBI
18	Ceballos G, Gutiérrez-Salmeán G and Meaney E: The vascular endothelium: A review series I. Basic aspects of the vascular endothelium. Rev Mex Cardiol. 26:95–100. 2015.
19	Zhang D, Wang W, Zhou D, Chen Y, Han L, Liu Y, Cao C, Zhao H and Liu G: Ghrelin inhibits apoptosis induced by palmitate in rat aortic endothelial cells. Med Sci Monit. 16:BR396–BR403. 2010.PubMed/NCBI
20	Yin S, Yang S, Pan X, Ma A, Ma J, Pei H, Dong Y, Li S, Li W and Bi X: MicroRNA155 promotes oxLDLinduced autophagy in human umbilical vein endothelial cells by targeting the PI3K/Akt/mTOR pathway. Mol Med Rep. 18:2798–2806. 2018.PubMed/NCBI
21	Kuhajda FP: Fatty acid synthase and cancer: New application of an old pathway. Cancer Res. 66:5977–5980. 2006. View Article : Google Scholar : PubMed/NCBI
22	Miura GI and Treisman JE: Lipid modification of secreted signaling proteins. Cell Cycle. 5:1184–1188. 2006. View Article : Google Scholar : PubMed/NCBI
23	Fiorentino M, Zadra G, Palescandolo E, Fedele G, Bailey D, Fiore C, Nguyen PL, Migita T, Zamponi R, Di Vizio D, et al: Overexpression of fatty acid synthase is associated with palmitoylation of Wnt1 and cytoplasmic stabilization of beta-catenin in prostate cancer. Lab Invest. 88:1340–1348. 2008. View Article : Google Scholar : PubMed/NCBI
24	Prossomariti A, Piazzi G, D'Angelo L, Miccoli S, Turchetti D, Alquati C, Montagna C, Bazzoli F and Ricciardiello L: miR-155 is downregulated in familial adenomatous polyposis and modulates WNT signaling by targeting AXIN1 and TCF4. Mol Cancer Res. 16:1965–1976. 2018. View Article : Google Scholar : PubMed/NCBI
25	Kong W, He L, Richards EJ, Challa S, Xu CX, Permuth-Wey J, Lancaster JM, Coppola D, Sellers TA, Djeu JY, et al: Upregulation of miRNA-155 promotes tumour angiogenesis by targeting VHL and is associated with poor prognosis and triple-negative breast cancer. Oncogene. 33:679–689. 2014. View Article : Google Scholar : PubMed/NCBI
26	Higashi Y, Noma K, Yoshizumi M and Kihara Y: Endothelial function and oxidative stress in cardiovascular diseases. Circ J. 73:411–418. 2009. View Article : Google Scholar : PubMed/NCBI
27	Miller TA, LeBrasseur NK, Cote GM, Trucillo MP, Pimentel DR, Ido Y, Ruderman NB and Sawyer DB: Oleate prevents palmitate-induced cytotoxic stress in cardiac myocytes. Biochem Biophys Res Commun. 336:309–315. 2005. View Article : Google Scholar : PubMed/NCBI
28	Namgaladze D, Lips S, Leiker TJ, Murphy RC, Ekroos K, Ferreiros N, Geisslinger G and Brune B: Inhibition of macrophage fatty acid β-oxidation exacerbates palmitate-induced inflammatory and endoplasmic reticulum stress responses. Diabetologia. 57:1067–1077. 2014. View Article : Google Scholar : PubMed/NCBI
29	Pillon NJ, Azizi PM, Li YE, Liu J, Wang C, Chan KL, Hopperton KE, Bazinet RP, Heit B, Bilan PJ, et al: Palmitate-induced inflammatory pathways in human adipose microvascular endothelial cells promote monocyte adhesion and impair insulin transcytosis. Am J Physiol Endocrinol Metab. 309:E35–E44. 2015. View Article : Google Scholar : PubMed/NCBI
30	Park K, Kim N, Nam J, Bang D and Lee ES: Association of TNFA promoter region haplotype in Behçet's disease. J Korean Med Sci. 21:596–601. 2006. View Article : Google Scholar : PubMed/NCBI
31	Kataoka H, Murakami R, Numaguchi Y, Okumura K and Murohara T: Angiotensin II type 1 receptor blockers prevent tumor necrosis factor-alpha-mediated endothelial nitric oxide synthase reduction and superoxide production in human umbilical vein endothelial cells. Eur J Pharmacol. 636:36–41. 2010. View Article : Google Scholar : PubMed/NCBI
32	Zhang H, Park Y, Wu J, Chen X, Lee S, Yang J, Dellsperger KC and Zhang C: Role of TNF-alpha in vascular dysfunction. Clin Sci (Lond). 116:219–230. 2009. View Article : Google Scholar : PubMed/NCBI
33	Zhang Y, Wei W, Cheng N, Wang K, Li B, Jiang X and Sun S: Hepatitis C virus-induced up-regulation of microRNA-155 promotes hepatocarcinogenesis by activating Wnt signaling. Hepatology. 56:1631–1640. 2012. View Article : Google Scholar : PubMed/NCBI