Inhibition of high glucose-induced apoptosis by uncoupling protein 2 in human umbilical vein endothelial cells

He,Ying; Wang,Ning; Shen,Yinchen; Zheng,Zhi; Xu,Xun

doi:10.3892/ijmm.2014.1676

Inhibition of high glucose-induced apoptosis by uncoupling protein 2 in human umbilical vein endothelial cells

Authors:
- Ying He
- Ning Wang
- Yinchen Shen
- Zhi Zheng
- Xun Xu
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Affiliations: Division of Opthalmology, Shanghai First People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200080, P.R. China
Published online on: February 27, 2014 https://doi.org/10.3892/ijmm.2014.1676
Pages: 1275-1281

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Abstract

Studies have shown that an overproduction of mitochondrial reactive oxygen species (ROS) is an initiating cause in the pathogenesis of diabetic complications. However, uncoupling protein 2 (UCP2) can protect retinal vascular endothelial cells from damage by inhibiting the overproduction of mitochondrial ROS, although the protective mechanism involved is not completely clear. This study aimed to assess the effect and mechanism of UCP2 on the apoptosis of human umbilical vein endothelial cells (HUVECs). HUVECs were cultured in normal glucose (NG, 5.5 mmol/l) or high glucose (HG, 30 mmol/l) medium in the presence or absence of UCP2+/+ lentiviral transfection. Lentivirus-mediated UCP2 overexpression inhibited the apoptosis of HUVECs induced by HG. Treatment with HG resulted in the upregulation of caspase-3 and cytochrome c and the downregulation of Bcl-2 in vitro. Furthermore, compared with the NG group, the rate of apoptosis was significantly increased in the HG group. On day two post-infection, NG cells showed significantly greater HUVEC cell proliferation than HG cells. Notably, UCP2 overexpression inhibited these processes. Taken together, these results suggest that UCP2 promotes cell proliferation and inhibits HG-induced apoptosis in HUVECs via the Bcl-2 up‑ and downregulation of caspase-3 and cytochrome c in vitro. This may provide experimental evidence for the application of UCP2 as a new protective factor for diabetic complications, such as diabetic retinopathy.

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1	Forbes JM, Coughlan MT and Cooper ME: Oxidative stress as a major culprit in kidney disease in diabetes. Diabetes. 57:1446–1454. 2008. View Article : Google Scholar : PubMed/NCBI
2	Brownlee M: Biochemistry and molecular cell biology of diabetic complications. Nature. 414:813–820. 2001. View Article : Google Scholar : PubMed/NCBI
3	Brownlee M: The pathobiology of diabetic complications: a unifying mechanism. Diabetes. 54:1615–1625. 2005. View Article : Google Scholar : PubMed/NCBI
4	Zheng Z, Chen H, Ke G, Fan Y, Zou H, et al: Protective effect of perindopril on diabetic retinopathy is associated with decreased vascular endothelial growth factor-to-pigment epithelium-derived factor ratio: involvement of a mitochondria-reactive oxygen species pathway. Diabetes. 58:954–964. 2009. View Article : Google Scholar
5	Mattson MP and Kroemer G: Mitochondria in cell death: novel targets for neuroprotection and cardioprotection. Trends Mol Med. 9:196–205. 2003. View Article : Google Scholar : PubMed/NCBI
6	Mattson MP and Liu D: Mitochondrial potassium channels and uncoupling proteins in synaptic plasticity and neuronal cell death. Biochem Biophys Res Commun. 304:539–549. 2003. View Article : Google Scholar : PubMed/NCBI
7	Cui Y, Xu X, Bi H, Wu J, et al: Expression modification of uncoupling proteins and MnSOD in retinal endothelial cells and pericytes induced by high glucose: the role of reactive oxygen species in diabetic retinopathy. Exp Eye Res. 83:807–816. 2001. View Article : Google Scholar
8	Fleury C, Neverova M, Collins S, Raimbault S, et al: Uncoupling protein-2: a novel gene linked to obesity and hyperinsulinemia. Nat Genet. 15:269–272. 1997. View Article : Google Scholar : PubMed/NCBI
9	Pecqueur C, Couplan E, Bouillaud F and Ricquier D: Genetic and physiological analysis of the role of uncoupling proteins in human energy homeostasis. J Mol Med. 79:48–56. 2001. View Article : Google Scholar
10	Pecqueur C, Alves-Guerra MC, Gelly C, Levi-Meyrueis C, Couplan E, et al: Uncoupling protein 2, in vivo distribution, induction upon oxidative stress, and evidence for translational regulation. J Biol Chem. 276:8705–8712. 2001. View Article : Google Scholar : PubMed/NCBI
11	Couplan E, del Mar Gonzalez-Barroso M, Alves-Guerra MC, Ricquier D, Goubern M and Bouillaud F: No evidence for a basal, retinoic, or superoxide-induced uncoupling activity of the uncoupling protein 2 present in spleen or lung mitochondria. J Biol Chem. 277:26268–26275. 2002. View Article : Google Scholar : PubMed/NCBI
12	Zhang CY, Baffy G, Perret P, Krauss S, Peroni O, et al: Uncoupling protein-2 negatively regulates insulin secretion and is a major link between obesity, beta cell dysfunction, and type 2 diabetes. Cell. 105:745–755. 2001. View Article : Google Scholar : PubMed/NCBI
13	Krauss S, Brand MD and Buttgereit F: Signaling takes a breath - new quantitative perspectives on bioenergetics and signal transduction. Immunity. 15:497–502. 2001. View Article : Google Scholar : PubMed/NCBI
14	Krauss S, Zhang CY, Scorrano L, Dalgaard LT, St-Pierre J, et al: Superoxide-mediated activation of uncoupling protein 2 causes pancreatic beta cell dysfunction. J Clin Invest. 112:1831–1842. 2003. View Article : Google Scholar : PubMed/NCBI
15	Turner JD, Gaspers LD, Wang G and Thomas AP: Uncoupling protein-2 modulates myocardial excitation-contraction coupling. Circ Res. 106:730–738. 2010. View Article : Google Scholar : PubMed/NCBI
16	Horvath TL, Warden CH, Hajos M, Lombardi A, Goglia F, et al: Brain uncoupling protein 2: uncoupled neuronal mitochondria predict thermal synapses in homeostatic centers. J Neurosci. 19:10417–10427. 1999.PubMed/NCBI
17	Liu Y, Chen L, Xu X, Vicaut E and Sercombe R: Both ischemic preconditioning and ghrelin administration protect hippocampus from ischemia/reperfusion and upregulate uncoupling protein-2. BMC Physiol. 9:172009. View Article : Google Scholar : PubMed/NCBI
18	Della-Morte D, Dave KR, DeFazio RA, Bao YC, Raval AP, et al: Resveratrol pretreatment protects rat brain from cerebral ischemic damage via a sirtuin 1-uncoupling protein 2 pathway. Neuroscience. 159:993–1002. 2009. View Article : Google Scholar
19	Arsenijevic D, Onuma H, Pecqueur C, Ricquier D, et al: Disruption of the uncoupling protein-2 gene in mice reveals a role in immunity and reactive oxygen species production. Nat Genet. 26:435–439. 2000. View Article : Google Scholar : PubMed/NCBI
20	Nègre-Salvayre A, Hirtz C, Carrera G, Casteilla L, et al: A role for uncoupling protein-2 as a regulator of mitochondrial hydrogen peroxide generation. FASEB J. 11:809–815. 1997.PubMed/NCBI
21	Li LX, Skorpen F, Egeberg K, Jørgensen IH and Grill V: Uncoupling protein-2 participates in cellular defense against oxidative stress in clonal beta-cells. Biochem Biophys Res Commun. 282:273–277. 2001. View Article : Google Scholar : PubMed/NCBI
22	Bai Y, Onuma H, Bai X, Medvedev AV, Collins S, et al: Persistent nuclear factor-kappa B activation in Ucp2−/− mice leads to enhanced nitric oxide and inflammatory cytokine production. J Biol Chem. 280:19062–19069. 2005.
23	Nishio K, Qiao S and Yamashita H: Characterization of the differential expression of uncoupling protein 2 and ROS production in differentiated mouse macrophage-cells (Mm1) and the progenitor cells (M1). J Mol Histol. 36:35–44. 2005. View Article : Google Scholar
24	Brand MD, Affourtit C, Esteves TC, Parker N, et al: Mitochondrial superoxide: production, biological effects, and activation of uncoupling proteins. Free Radic Biol Med. 37:755–767. 2004. View Article : Google Scholar : PubMed/NCBI
25	Conti B, Sugama S, Lucero J, Bartfai T, et al: Uncoupling protein 2 protects dopaminergic neurons from acute 1,2,3,6-methyl-phenyl-tetrahydropyridine toxicity. J Neurochem. 93:493–501. 2005. View Article : Google Scholar
26	Mattiasson G, Shamloo M, Gido G, Wieloch T, et al: Uncoupling protein-2 prevents neuronal death and diminishes brain dysfunction after stroke and brain trauma. Nat Med. 9:1062–1068. 2003. View Article : Google Scholar : PubMed/NCBI
27	Paradis E, Clavel S, Bouillaud F, Ricquier D and Richard D: Uncoupling protein 2: a novel player in neuroprotection. Trends Mol Med. 9:522–525. 2003. View Article : Google Scholar : PubMed/NCBI
28	Richard D, Clavel S, Huang Q, Sanchis D and Ricquier D: Uncoupling protein 2 in the brain: distribution and function. Biochem Soc Trans. 29:812–817. 2001. View Article : Google Scholar : PubMed/NCBI
29	Blanc J, Alves-Guerra MC, Esposito B, Rousset S, Mallat Z, et al: Protective role of uncoupling protein 2 in atherosclerosis. Circulation. 107:388–390. 2003. View Article : Google Scholar : PubMed/NCBI
30	Teshima Y, Akao M, Jones SP and Marbán E: Uncoupling protein-2 overexpression inhibits mitochondrial death pathway in cardiomyocytes. Circ Res. 93:192–200. 2003. View Article : Google Scholar : PubMed/NCBI
31	Polonsky KS and Semenkovich CF: The pancreatic beta cell heats up: UCP2 and insulin secretion in diabetes. Cell. 105:705–707. 2001. View Article : Google Scholar : PubMed/NCBI
32	Suh YH, Kim SY, Lee HY, Song DK, et al: Overexpression of short heterodimer partner recovers impaired glucose-stimulated insulin secretion of pancreatic beta-cells overexpressing UCP2. J Endocrinol. 183:133–144. 2004. View Article : Google Scholar
33	Sayeed A, Meng Z, Luciani G, Dairkee SH, et al: Negative regulation of UCP2 by TGFβ signaling characterizes low and intermediate-grade primary breast cancer. Cell Death Dis. 1:e532010.
34	Beltramo E, Berrone E, Buttiglieri S and Porta M: Thiamine and benfotiamine prevent increased apoptosis in endothelial cells and pericytes cultured in high glucose. Diabetes Metab Res Rev. 20:330–336. 2004. View Article : Google Scholar : PubMed/NCBI
35	Li W, Liu X, He Z, Yanoff M, Jian B and Ye X: Expression of apoptosis regulatory genes by retinal pericytes after rapid glucose reduction. Invest Ophthalmol Vis Sci. 39:1535–1543. 1998.PubMed/NCBI
36	Naruse K, Nakamura J, Hamada Y, Nakayama M, Chava S, Komori T, Kato K, Kasuya Y, Miwa K and Hotta N: Aldose reductase inhibition prevents glucose-induced apoptosis in cultured bovine retinal microvascular pericytes. Exp Eye Res. 71:309–315. 2000. View Article : Google Scholar
37	Romeo G, Liu WH, Asnaghi V, Kern TS and Lorenzi M: Activation of nuclear factor-kappaB induced by diabetes and high glucose regulates a proapoptotic program in retinal pericytes. Diabetes. 51:2241–2248. 2002. View Article : Google Scholar : PubMed/NCBI
38	Elorza A, Hyde B, Mikkola HK, Collins S and Shirihai OS: UCP2 modulates cell proliferation through the MAPK/ERK pathway during erythropoiesis and has no effect on heme biosynthesis. J Biol Chem. 283:30461–30470. 2008. View Article : Google Scholar : PubMed/NCBI
39	Pecqueur C, Bui T, Gelly C, Tompson CB, et al: Uncoupling protein-2 controls proliferation by promoting fatty acid oxidation and limiting glycolysis-derived pyruvate utilization. FASEB J. 22:9–18. 2008. View Article : Google Scholar : PubMed/NCBI
40	Sears RC and Nevins JR: Signaling networks that link cell proliferation and cell fate. J Biol Chem. 277:11617–11620. 2002. View Article : Google Scholar : PubMed/NCBI