TRB3 mediates advanced glycation end product-induced apoptosis of pancreatic β-cells through the protein kinase C β pathway

Wang,Meng; Zhang,Wenjian; Xu,Shiqing; Peng,Liang; Wang,Zai; Liu,Honglin; Fang,Qing; Deng,Tingting; Men,Xiuli; Lou,Jinning

doi:10.3892/ijmm.2017.2991

TRB3 mediates advanced glycation end product-induced apoptosis of pancreatic β-cells through the protein kinase C β pathway

Authors:
- Meng Wang
- Wenjian Zhang
- Shiqing Xu
- Liang Peng
- Zai Wang
- Honglin Liu
- Qing Fang
- Tingting Deng
- Xiuli Men
- Jinning Lou
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Affiliations: Graduate School of Peking Union Medical College, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100730, P.R. China, Institute of Clinical Medical Sciences, China-Japan Friendship Hospital, Beijing 100029, P.R. China, Department of Pathophysiology, North China University of Science and Technology, Tangshan, Hebei 063000, P.R. China
Published online on: May 16, 2017 https://doi.org/10.3892/ijmm.2017.2991
Pages: 130-136
Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Advanced glycation end products (AGEs), which accumulate in the body during the development of diabetes, may be one of the factors leading to pancreatic β-cell failure and reduced β-cell mass. However, the mechanisms responsible for AGE‑induced apoptosis remain unclear. This study identified the role and mechanisms of action of tribbles homolog 3 (TRB3) in AGE-induced β-cell oxidative damage and apoptosis. Rat insulinoma cells (INS-1) were treated with 200 µg/ml AGEs for 48 h, and cell apoptosis was then detected by TUNEL staining and flow cytometry. The level of intracellular reactive oxygen species (ROS) was measured by a fluorescence assay. The expression levels of receptor of AGEs (RAGE), TRB3, protein kinase C β2 (PKCβ2) and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 4 (NOX4) were evaluated by RT-qPCR and western blot analysis. siRNA was used to knockdown TRB3 expression through lipofection, followed by an analysis of the effects of TRB3 on PKCβ2 and NOX4. Furthermore, the PKCβ2-specific inhibitor, LY333531, was used to analyze the effects of PKCβ2 on ROS levels and apoptosis. We found that AGEs induced the apoptosis of INS-1 cells and upregulated RAGE and TRB3 expression. AGEs also increased ROS levels in β-cells. Following the knockdown of TRB3, the AGE-induced apoptosis and intracellular ROS levels were significantly decreased, suggesting that TRB3 mediated AGE-induced apoptosis. Further experiments demonstrated that the knockdown of TRB3 decreased the PKCβ2 and NOX4 expression levels. When TRB3 was knocked down, the cells expressed decreased levels of PKCβ2 and NOX4. The PKCβ2‑specific inhibitor, LY333531, also reduced AGE-induced apoptosis and intracellular ROS levels. Taken together, our data suggest that TRB3 mediates AGE-induced oxidative injury in β-cells through the PKCβ2 pathway.

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1	Nowotny K, Jung T, Höhn A, Weber D and Grune T: Advanced glycation end products and oxidative stress in type 2 diabetes mellitus. Biomolecules. 5:194–222. 2015. View Article : Google Scholar : PubMed/NCBI
2	Volchuk A and Ron D: The endoplasmic reticulum stress response in the pancreatic β-cell. Diabetes Obes Metab. 12(Suppl 2): 48–57. 2010. View Article : Google Scholar
3	Lim M, Park L, Shin G, Hong H, Kang I and Park Y: Induction of apoptosis of β cells of the pancreas by advanced glycation end-products, important mediators of chronic complications of diabetes mellitus. Ann N Y Acad Sci. 1150:311–315. 2008. View Article : Google Scholar
4	Lee BW, Chae HY, Kwon SJ, Park SY, Ihm J and Ihm SH: RAGE ligands induce apoptotic cell death of pancreatic β-cells via oxidative stress. Int J Mol Med. 26:813–818. 2010.PubMed/NCBI
5	Jung H, Joo J, Jeon Y, Lee J, In J, Kim D, Kang E, Kim Y, Lim Y, Kang J, et al: Advanced glycation end products downregulate glucokinase in mice. Diabetes Metab Res Rev. 27:557–563. 2011. View Article : Google Scholar : PubMed/NCBI
6	Puddu A, Storace D, Durante A, Odetti P and Viviani GL: Glucagon-like peptide-1 counteracts the detrimental effects of advanced glycation end-products in the pancreatic beta cell line HIT-T 15. Biochem Biophys Res Commun. 398:462–466. 2010. View Article : Google Scholar : PubMed/NCBI
7	Jakus V and Rietbrock N: Advanced glycation end-products and the progress of diabetic vascular complications. Physiol Res. 53:131–142. 2004.PubMed/NCBI
8	Baumann M: Role of advanced glycation end products in hypertension and cardiovascular risk: human studies. J Am Soc Hypertens. 6:427–435. 2012. View Article : Google Scholar : PubMed/NCBI
9	Chilelli NC, Burlina S and Lapolla A: AGEs, rather than hyperglycemia, are responsible for microvascular complications in diabetes: a 'glycoxidation-centric' point of view. Nutr Metab Cardiovasc Dis. 23:913–919. 2013. View Article : Google Scholar : PubMed/NCBI
10	Yamagishi S, Nakamura N, Suematsu M, Kaseda K and Matsui T: Advanced glycation end products: a molecular target for vascular complications in diabetes. Mol Med. 21(Suppl 1): S32–S40. 2015. View Article : Google Scholar : PubMed/NCBI
11	Li BY, Li XL, Gao HQ, Zhang JH, Cai Q, Cheng M and Lu M: Grape seed procyanidin B2 inhibits advanced glycation end product-induced endothelial cell apoptosis through regulating GSK3β phosphorylation. Cell Biol Int. 35:663–669. 2011. View Article : Google Scholar : PubMed/NCBI
12	Sun C, Liang C, Ren Y, Zhen Y, He Z, Wang H, Tan H, Pan X and Wu Z: Advanced glycation end products depress function of endothelial progenitor cells via p38 and ERK 1/2 mitogen-activated protein kinase pathways. Basic Res Cardiol. 104:42–49. 2009. View Article : Google Scholar
13	Adamopoulos C, Piperi C, Gargalionis AN, Dalagiorgou G, Spilioti E, Korkolopoulou P, Diamanti-Kandarakis E and Papavassiliou AG: Advanced glycation end products upregulate lysyl oxidase and endothelin-1 in human aortic endothelial cells via parallel activation of ERK1/2-NF-κB and JNK-AP-1 signaling pathways. Cell Mol Life Sci. 73:1685–1698. 2016. View Article : Google Scholar
14	Morita M, Yano S, Yamaguchi T and Sugimoto T: Advanced glycation end products-induced reactive oxygen species generation is partly through NF-kappa B activation in human aortic endothelial cells. J Diabetes Complications. 27:11–15. 2013. View Article : Google Scholar
15	Sang HQ, Gu JF, Yuan JR, Zhang MH, Jia XB and Feng L: The protective effect of Smilax glabra extract on advanced glycation end products-induced endothelial dysfunction in HUVECs via RAGE-ERK1/2-NF-κB pathway. J Ethnopharmacol. 155:785–795. 2014. View Article : Google Scholar : PubMed/NCBI
16	Lin N, Zhang H and Su Q: Advanced glycation end-products induce injury to pancreatic beta cells through oxidative stress. Diabetes Metab. 38:250–257. 2012. View Article : Google Scholar : PubMed/NCBI
17	Zhu Y, Shu T, Lin Y, Wang H, Yang J, Shi Y and Han X: Inhibition of the receptor for advanced glycation endproducts (RAGE) protects pancreatic β-cells. Biochem Biophys Res Commun. 404:159–165. 2011. View Article : Google Scholar
18	Coughlan MT, Yap FY, Tong DC, Andrikopoulos S, Gasser A, Thallas-Bonke V, Webster DE, Miyazaki J, Kay TW, Slattery RM, et al: Advanced glycation end products are direct modulators of β-cell function. Diabetes. 60:2523–2532. 2011. View Article : Google Scholar : PubMed/NCBI
19	Cheng WP, Wang BW, Lo HM and Shyu KG: Mechanical stretch induces apoptosis regulator TRB3 in cultured cardiomyocytes and volume-overloaded heart. PLoS One. 10:e01232352015. View Article : Google Scholar : PubMed/NCBI
20	Du K, Herzig S, Kulkarni RN and Montminy M: TRB3: a tribbles homolog that inhibits Akt/PKB activation by insulin in liver. Science. 300:1574–1577. 2003. View Article : Google Scholar : PubMed/NCBI
21	Qian B, Wang H, Men X, Zhang W, Cai H, Xu S, Xu Y, Ye L, Wollheim CB and Lou J: TRIB3 [corrected] is implicated in glucotoxicity- and endoplasmic reticulum-stress-induced [corrected] beta-cell apoptosis. J Endocrinol. 199:407–416. 2008. View Article : Google Scholar : PubMed/NCBI
22	Qin J, Fang N, Lou J, Zhang W, Xu S, Liu H, Fang Q, Wang Z, Liu J, Men X, et al: TRB3 is involved in free fatty acid-induced INS-1-derived cell apoptosis via the protein kinase C δ pathway. PLoS One. 9:e960892014. View Article : Google Scholar
23	Fang N, Zhang W, Xu S, Lin H, Wang Z, Liu H, Fang Q, Li C, Peng L and Lou J: TRIB3 alters endoplasmic reticulum stress-induced β-cell apoptosis via the NF-κB pathway. Metabolism. 63:822–830. 2014. View Article : Google Scholar : PubMed/NCBI
24	Scivittaro V, Ganz MB and Weiss MF: AGEs induce oxidative stress and activate protein kinase C-β(II) in neonatal mesangial cells. Am J Physiol Renal Physiol. 278:F676–F683. 2000.PubMed/NCBI
25	Li L and Renier G: Activation of nicotinamide adenine dinucleotide phosphate (reduced form) oxidase by advanced glycation end products links oxidative stress to altered retinal vascular endothelial growth factor expression. Metabolism. 55:1516–1523. 2006. View Article : Google Scholar : PubMed/NCBI
26	Wang H, Jiang YW, Zhang WJ, Xu SQ, Liu HL, Yang WY and Lou JN: Differential activations of PKC/PKA related to microvasculopathy in diabetic GK rats. Am J Physiol Endocrinol Metab. 302:E173–E182. 2012. View Article : Google Scholar
27	Zhang L, Huang D, Shen D, Zhang C, Ma Y, Babcock SA, Chen B and Ren J: Inhibition of protein kinase C βII isoform ameliorates methylglyoxal advanced glycation endproduct-induced cardiomyocyte contractile dysfunction. Life Sci. 94:83–91. 2014. View Article : Google Scholar
28	Guo B, Zhang W, Xu S, Lou J, Wang S and Men X: GSK-3β mediates dexamethasone-induced pancreatic β cell apoptosis. Life Sci. 144:1–7. 2016. View Article : Google Scholar
29	Singh VP, Bali A, Singh N and Jaggi AS: Advanced glycation end products and diabetic complications. Korean J Physiol Pharmacol. 18:1–14. 2014. View Article : Google Scholar : PubMed/NCBI
30	Bierhaus A, Humpert PM, Morcos M, Wendt T, Chavakis T, Arnold B, Stern DM and Nawroth PP: Understanding RAGE, the receptor for advanced glycation end products. J Mol Med (Berl). 83:876–886. 2005. View Article : Google Scholar
31	Neeper M, Schmidt AM, Brett J, Yan SD, Wang F, Pan YC, Elliston K, Stern D and Shaw A: Cloning and expression of a cell surface receptor for advanced glycosylation end products of proteins. J Biol Chem. 267:14998–15004. 1992.PubMed/NCBI
32	Hofmann MA, Drury S, Fu C, Qu W, Taguchi A, Lu Y, Avila C, Kambham N, Bierhaus A, Nawroth P, et al: RAGE mediates a novel proinflammatory axis: a central cell surface receptor for S100/calgranulin polypeptides. Cell. 97:889–901. 1999. View Article : Google Scholar : PubMed/NCBI
33	Hori O, Brett J, Slattery T, Cao R, Zhang J, Chen JX, Nagashima M, Lundh ER, Vijay S, Nitecki D, et al: The receptor for advanced glycation end products (RAGE) is a cellular binding site for amphoterin. Mediation of neurite outgrowth and co-expression of rage and amphoterin in the developing nervous system. J Biol Chem. 270:25752–25761. 1995. View Article : Google Scholar : PubMed/NCBI
34	Deane R, Du Yan S, Submamaryan RK, LaRue B, Jovanovic S, Hogg E, Welch D, Manness L, Lin C, Yu J, et al: RAGE mediates amyloid-β peptide transport across the blood-brain barrier and accumulation in brain. Nat Med. 9:907–913. 2003. View Article : Google Scholar : PubMed/NCBI
35	Mackic JB, Stins M, McComb JG, Calero M, Ghiso J, Kim KS, Yan SD, Stern D, Schmidt AM, Frangione B, et al: Human blood-brain barrier receptors for Alzheimer's amyloid-beta 1–;40. Asymmetrical binding, endocytosis, and transcytosis at the apical side of brain microvascular endothelial cell monolayer. J Clin Invest. 102:734–743. 1998. View Article : Google Scholar : PubMed/NCBI
36	Lo MC, Chen MH, Lee WS, Lu CI, Chang CR, Kao SH and Lee HM: Nε-(carboxymethyl) lysine-induced mitochondrial fission and mitophagy cause decreased insulin secretion from β-cells. Am J Physiol Endocrinol Metab. 309:E829–E839. 2015.PubMed/NCBI
37	Hasnain SZ, Prins JB and McGuckin MA: Oxidative and endoplasmic reticulum stress in β-cell dysfunction in diabetes. J Mol Endocrinol. 56:R33–R54. 2016. View Article : Google Scholar
38	Evans JL, Goldfine ID, Maddux BA and Grodsky GM: Are oxidative stress-activated signaling pathways mediators of insulin resistance and β-cell dysfunction? Diabetes. 52:1–8. 2003. View Article : Google Scholar
39	Nisimoto Y, Diebold BA, Cosentino-Gomes D and Lambeth JD: Nox4: a hydrogen peroxide-generating oxygen sensor. Biochemistry. 53:5111–5120. 2014. View Article : Google Scholar : PubMed/NCBI
40	Prudente S, Sesti G, Pandolfi A, Andreozzi F, Consoli A and Trischitta V: The mammalian tribbles homolog TRIB3, glucose homeostasis, and cardiovascular diseases. Endocr Rev. 33:526–546. 2012. View Article : Google Scholar : PubMed/NCBI
41	Humphrey RK, Newcomb CJ, Yu SM, Hao E, Yu D, Krajewski S, Du K and Jhala US: Mixed lineage kinase-3 stabilizes and functionally cooperates with TRIBBLES-3 to compromise mitochondrial integrity in cytokine-induced death of pancreatic beta cells. J Biol Chem. 285:22426–22436. 2010. View Article : Google Scholar : PubMed/NCBI
42	Ohoka N, Yoshii S, Hattori T, Onozaki K and Hayashi H: TRB3, a novel ER stress-inducible gene, is induced via ATF4-CHOP pathway and is involved in cell death. EMBO J. 24:1243–1255. 2005. View Article : Google Scholar : PubMed/NCBI
43	Zou CG, Cao XZ, Zhao YS, Gao SY, Li SD, Liu XY, Zhang Y and Zhang KQ: The molecular mechanism of endoplasmic reticulum stress-induced apoptosis in PC-12 neuronal cells: the protective effect of insulin-like growth factor I. Endocrinology. 150:277–285. 2009. View Article : Google Scholar
44	Schwarzer R, Dames S, Tondera D, Klippel A and Kaufmann J: TRB3 is a PI 3-kinase dependent indicator for nutrient starvation. Cell Signal. 18:899–909. 2006. View Article : Google Scholar
45	Ord D, Meerits K and Ord T: TRB3 protects cells against the growth inhibitory and cytotoxic effect of ATF4. Exp Cell Res. 313:3556–3567. 2007. View Article : Google Scholar : PubMed/NCBI
46	Gorasia DG, Dudek L, Veith PD, Shankar R, Safavi-Hemami H, Williamson NA, Reynolds EC, Hubbard MJ and Purcell AW: Pancreatic beta cells are highly susceptible to oxidative and ER stresses during the development of diabetes. J Proteome Res. 14:688–699. 2015. View Article : Google Scholar
47	Zhang K: Integration of ER stress, oxidative stress and the inflammatory response in health and disease. Int J Clin Exp Med. 3:33–40. 2010.PubMed/NCBI
48	Pérez LM, Milkiewicz P, Ahmed-Choudhury J, Elias E, Ochoa JE, Sánchez Pozzi EJ, Coleman R and Roma MG: Oxidative stress induces actin-cytoskeletal and tight-junctional alterations in hepatocytes by a Ca²⁺-dependent, PKC-mediated mechanism: protective effect of PKA. Free Radic Biol Med. 40:2005–2017. 2006. View Article : Google Scholar
49	Pérez LM, Milkiewicz P, Elias E, Coleman R, Sánchez Pozzi EJ and Roma MG: Oxidative stress induces internalization of the bile salt export pump, Bsep, and bile salt secretory failure in isolated rat hepatocyte couplets: a role for protein kinase C and prevention by protein kinase A. Toxicol Sci. 91:150–158. 2006. View Article : Google Scholar : PubMed/NCBI