Phosphorylation of signal transducer and activator of transcription 3 induced by hyperglycemia is different with 
that induced by lipopolysaccharide or erythropoietin via receptor‑coupled signaling in cardiac cells

Chiu,Yu‑Hsin; Ku,Po‑Ming; Cheng,Yung‑Ze; Li,Yingxiao; Cheng,Juei‑Tang; Niu,Ho‑Shan

doi:10.3892/mmr.2017.7973

Phosphorylation of signal transducer and activator of transcription 3 induced by hyperglycemia is different with that induced by lipopolysaccharide or erythropoietin via receptor‑coupled signaling in cardiac cells

Authors:
- Yu‑Hsin Chiu
- Po‑Ming Ku
- Yung‑Ze Cheng
- Yingxiao Li
- Juei‑Tang Cheng
- Ho‑Shan Niu
View Affiliations

Affiliations: Division of Infectious Diseases, Chi‑Mei Medical Center‑Liouying, Tainan 73601, Taiwan, R.O.C. , Cardiovascular Center, Department of Internal Medicine, Chi‑Mei Medical Center‑Liouying, Tainan 73601, Taiwan, R.O.C. , Department of Emergency Medicine, Chi‑Mei Medical Center, Tainan 71003, Taiwan, R.O.C., Department of Medical Research, Chi‑Mei Medical Center, Tainan 71003, Taiwan, R.O.C., Department of Nursing, Tzu Chi University of Science and Technology, Hualien 97005, Taiwan, R.O.C.
Published online on: November 6, 2017 https://doi.org/10.3892/mmr.2017.7973
Pages: 1311-1320

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 signal transducer and activator of transcription 3 (STAT3) is known to be involved in hypertrophy and fibrosis in cardiac dysfunction. The activation of STAT3 via the phosphorylation of STAT3 is required for the production of functional activity. It has been established that lipopolysaccharide (LPS)‑induced phosphorylation of STAT3 in cardiomyocytes primarily occurs through a direct receptor‑mediated action. This effect is demonstrated to be produced rapidly. STAT3 in cardiac fibrosis of diabetes is induced by high glucose through promotion of the STAT3‑associated signaling pathway. However, the time schedule for STAT3 activation between LPS and high glucose appears to be different. Therefore, the difference in STAT3 activation between LPS and hyperglycemia in cardiomyocytes requires elucidation. The present study investigated the phosphorylation of STAT3 induced by LPS and hyperglycemia in the rat cardiac cell line H9c2. Additionally, phosphorylation of STAT3 induced by erythropoietin (EPO) via receptor activation was compared. Then, the downstream signals for fibrosis, including the connective tissue growth factor (CTGF) and matrix metalloproteinase (MMP)‑9, were determined using western blotting, while the mRNA levels were quantified. LPS induced a rapid elevation of STAT3 phosphorylation in H9c2 cells within 30 min, similar to that produced by EPO. However, LPS or EPO failed to modify the mRNA level of STAT3, and/or the downstream signals for fibrosis. High glucose increased STAT3 phosphorylation to be stable after a long period of incubation. Glucose incubation for 24 h may augment the STAT3 expression in a dose‑dependent manner. Consequently, fibrosis‑associated signals, including CTGF and MMP‑9 protein, were raised in parallel. In the presence of tiron, an antioxidant, these changes by hyperglycemia were markedly reduced, demonstrating the mediation of oxidative stress. Therefore, LPS‑ or EPO‑induced STAT3 phosphorylation is different compared with that caused by high glucose in H9c2 cells. Sustained activation of STAT3 by hyperglycemia may promote the expression of fibrosis‑associated signals, including CTGF and MMP‑9, in H9c2 cells. Therefore, regarding the cardiac dysfunctions associated with diabetes and/or hyperglycemia, the identification of nuclear STAT3 may be more reliable compared with the assay of phosphorylated STAT3 in cardiac cells.

View Figures

View References

1	Haghikia A, Ricke-Hoch M, Stapel B, Gorst I and Hilfiker-Kleiner D: STAT3, a key regulator of cell-to-cell communication in the heart. Cardiovasc Res. 102:281–289. 2014. View Article : Google Scholar : PubMed/NCBI
2	Boengler K, Hilfiker-Kleiner D, Heusch G and Schulz R: Inhibition of permeability transition pore opening by mitochondrial STAT3 and its role in myocardial ischemia/reperfusion. Basic Res Cardiol. 105:771–785. 2010. View Article : Google Scholar : PubMed/NCBI
3	Elschami M, Scherr M, Philippens B and Gerardy-Schahn R: Reduction of STAT3 expression induces mitochondrial dysfunction and autophagy in cardiac HL-1 cells. Eur J Cell Biol. 92:21–29. 2013. View Article : Google Scholar : PubMed/NCBI
4	Heusch G, Musiolik J, Gedik N and Skyschally A: Mitochondrial STAT3 activation and cardioprotection by ischemic postconditioning in pigs with regional myocardial ischemia/reperfusion. Circ Res. 109:1302–1308. 2011. View Article : Google Scholar : PubMed/NCBI
5	Wegrzyn J, Potla R, Chwae YJ, Sepuri NB, Zhang Q, Koeck T, Derecka M, Szczepanek K, Szelag M, Gornicka A, et al: Function of mitochondrial Stat3 in cellular respiration. Science. 323:793–797. 2009. View Article : Google Scholar : PubMed/NCBI
6	Haghikia A, Stapel B, Hoch M and Hilfiker-Kleiner D: STAT3 and cardiac remodeling. Heart Fail Rev. 16:35–47. 2011. View Article : Google Scholar : PubMed/NCBI
7	Hilfiker-Kleiner D, Hilfiker A, Fuchs M, Kaminski K, Schaefer A, Schieffer B, Hillmer A, Schmiedl A, Ding Z, Podewski E, et al: Signal transducer and activator of transcription 3 is required for myocardial capillary growth, control of interstitial matrix deposition and heart protection from ischemic injury. Circ Res. 95:187–195. 2004. View Article : Google Scholar : PubMed/NCBI
8	Zouein FA, Kurdi M and Booz GW: Dancing rhinos in stilettos: The amazing saga of the genomic and nongenomic actions of STAT3 in the heart. JAKSTAT. 2:e243522013.PubMed/NCBI
9	Wang S, Li B, Li C, Cui W and Miao L: Potential renoprotective agents through inhibiting CTGF/CCN2 in diabetic nephropathy. J Diabetes Res. 2015:9623832015. View Article : Google Scholar : PubMed/NCBI
10	Miller AM, Wang H, Bertola A, Park O, Horiguchi N, Ki SH, Yin S, Lafdil F and Gao B: Inflammation-associated interleukin-6/signal transducer and activator of transcription 3 activation ameliorates alcoholic and nonalcoholic fatty liver diseases in interleukin-10-deficient mice. Hepatology. 54:846–856. 2011. View Article : Google Scholar : PubMed/NCBI
11	Jung JE, Lee HG, Cho IH, Chung DH, Yoon SH, Yang YM, Lee JW, Choi S, Park JW, Ye SK and Chung MH: STAT3 is a potential modulator of HIF-1-mediated VEGF expression in human renal carcinoma cells. FASEB J. 19:1296–1298. 2005.PubMed/NCBI
12	Chen Y, Wang JJ, Li J, Hosoya KI, Ratan R, Townes T and Zhang SX: Activating transcription factor 4 mediates hyperglycaemia-induced endothelial inflammation and retinal vascular leakage through activation of STAT3 in a mouse model of type 1 diabetes. Diabetologia. 55:2533–2545. 2012. View Article : Google Scholar : PubMed/NCBI
13	Saengboonmee C, Seubwai W, Pairojkul C and Wongkham S: High glucose enhances progression of cholangiocarcinoma cells via STAT3 activation. Sci Rep. 6:189952016. View Article : Google Scholar : PubMed/NCBI
14	Liu L, McBride KM and Reich NC: STAT3 nuclear import is independent of tyrosine phosphorylation and mediated by importin-alpha3. Proc Natl Acad Sci USA. 102:8150–8155. 2005. View Article : Google Scholar : PubMed/NCBI
15	Yang J, Chatterjee-Kishore M, Staugaitis SM, Nguyen H, Schlessinger K, Levy DE and Stark GR: Novel roles of unphosphorylated STAT3 in oncogenesis and transcriptional regulation. Cancer Res. 65:939–947. 2005.PubMed/NCBI
16	Meldrum DR: Tumor necrosis factor in the heart. Am J Physiol. 274:R577–R595. 1998.PubMed/NCBI
17	Wagner DR, McTiernan C, Sanders VJ and Feldman AM: Adenosine inhibits lipopolysaccharide-induced secretion of tumor necrosis factor-alpha in the failing human heart. Circulation. 97:521–524. 1998. View Article : Google Scholar : PubMed/NCBI
18	Cowan DB, Poutias DN, Del Nido PJ and McGowan FX Jr: CD14-independent activation of cardiomyocyte signal transduction by bacterial endotoxin. Am J Physiol Heart Circ Physiol. 279:H619–H629. 2000.PubMed/NCBI
19	Darnell JE Jr: STATs and gene regulation. Science. 277:1630–1635. 1997. View Article : Google Scholar : PubMed/NCBI
20	Ruff-Jamison S, Zhong Z, Wen Z, Chen K, Darnell JE Jr and Cohen S: Epidermal growth factor and lipopolysaccharide activate Stat3 transcription factor in mouse liver. J Biol Chem. 269:21933–21935. 1994.PubMed/NCBI
21	Chow JC, Young DW, Golenbock DT, Christ WJ and Gusovsky F: Toll-like receptor-4 mediates lipopolysaccharide-induced signal transduction. J Biol Chem. 274:10689–10692. 1999. View Article : Google Scholar : PubMed/NCBI
22	Avlas O, Fallach R, Shainberg A, Porat E and Hochhauser E: Toll-like receptor 4 stimulation initiates an inflammatory response that decreases cardiomyocyte contractility. Antioxid Redox Signal. 15:1895–1909. 2011. View Article : Google Scholar : PubMed/NCBI
23	Seavey MM and Dobrzanski P: The many faces of Janus kinase. Biochem Pharmacol. 83:1136–1145. 2012. View Article : Google Scholar : PubMed/NCBI
24	Xiao L, Li Z, Xu P, Li Z, Xu J and Yang Z: The expression of EPOR in renal cortex during postnatal development. PloS one. 7:e419932012. View Article : Google Scholar : PubMed/NCBI
25	Yu X, Shacka JJ, Eells JB, Suarez-Quian C, Przygodzki RM, Beleslin-Cokic B, Lin CS, Nikodem VM, Hempstead B, Flanders KC, et al: Erythropoietin receptor signalling is required for normal brain development. Development. 129:505–516. 2002.PubMed/NCBI
26	Qiao S, Mao X, Wang Y, Lei S, Liu Y, Wang T, Wong GT, Cheung CW, Xia Z, Irwin MG, et al: Remifentanil preconditioning reduces postischemic myocardial infarction and improves left ventricular performance via activation of the janus activated kinase-2/signal transducers and activators of transcription-3 signal pathway and subsequent inhibition of glycogen synthase kinase-3beta in rats. Crit Care Med. 44:e131–e145. 2016. View Article : Google Scholar : PubMed/NCBI
27	Fenton MJ and Golenbock DT: LPS-binding proteins and receptors. J Leukoc Biol. 64:25–32. 1998.PubMed/NCBI
28	Dai B, Cui M, Zhu M, Su WL, Qiu MC and Zhang H: STAT1/3 and ERK1/2 synergistically regulate cardiac fibrosis induced by high glucose. J Leukoc Biol. 32:960–971. 2013.
29	Fiaschi T, Magherini F, Gamberi T, Lucchese G, Faggian G, Modesti A and Modesti PA: Hyperglycemia and angiotensin II cooperate to enhance collagen I deposition by cardiac fibroblasts through a ROS-STAT3-dependent mechanism. Biochim Biophys Acta. 1843:2603–2610. 2014. View Article : Google Scholar : PubMed/NCBI
30	Wu L, Tan JL, Wang ZH, Chen YX, Gao L, Liu JL, Shi YH, Endoh M and Yang HT: ROS generated during early reperfusion contribute to intermittent hypobaric hypoxia-afforded cardioprotection against postischemia-induced Ca(2+) overload and contractile dysfunction via the JAK2/STAT3 pathway. J Mol Cell Cardiol. 81:150–161. 2015. View Article : Google Scholar : PubMed/NCBI
31	Watkins SJ, Borthwick GM and Arthur HM: The H9C2 cell line and primary neonatal cardiomyocyte cells show similar hypertrophic responses in vitro. In Vitro Cell Dev Biol Anim. 47:125–131. 2011. View Article : Google Scholar : PubMed/NCBI
32	Fan MJ, Huang-Liu R, Shen CY, Ju DT, Lin YM, Pai P, Huang PY, Ho TJ, Tsai FJ, Tsai CH and Huang CY: Reduction of TLR4 mRNA stability and protein expressions through inhibiting cytoplasmic translocation of HuR transcription factor by E₂ and/or ERα in LPS-treated H9c2 cardiomyoblast cells. Chin J Physiol. 57:8–18. 2014. View Article : Google Scholar : PubMed/NCBI
33	Parvin A, Pranap R, Shalini U, Devendran A, Baker JE and Dhanasekaran A: Erythropoietin protects cardiomyocytes from cell death during hypoxia/reperfusion injury through activation of survival signaling pathways. PLoS One. 9:e1074532014. View Article : Google Scholar : PubMed/NCBI
34	Yang L, Luo C, Chen C, Wang X, Shi W and Liu J: All-trans retinoic acid protects against doxorubicin-induced cardiotoxicity by activating the ERK2 signalling pathway. Br J Pharmacol. 173:357–371. 2016. View Article : Google Scholar : PubMed/NCBI
35	Mar GY, Ku PM, Chen LJ, Cheng KC, Li YX and Cheng JT: Increase in cardiac M2-muscarinic receptor expression is regulated by GATA binding protein 4 (GATA-4) in streptozotocin-induced diabetic rats. Int J Cardiol. 167:436–441. 2013. View Article : Google Scholar : PubMed/NCBI
36	Cheng YZ, Chen LJ, Lee WJ, Chen MF, Jung Lin H and Cheng JT: Increase of myocardial performance by rhodiola-ethanol extract in diabetic rats. J Ethnopharmacol. 144:234–239. 2012. View Article : Google Scholar : PubMed/NCBI
37	Cheng JT, Yu BC and Tong YC: Changes of M3-muscarinic receptor protein and mRNA expressions in the bladder urothelium and muscle layer of streptozotocin-induced diabetic rats. Neurosci Lett. 423:1–5. 2007. View Article : Google Scholar : PubMed/NCBI
38	Carraway MS, Suliman HB, Jones WS, Chen CW, Babiker A and Piantadosi CA: Erythropoietin activates mitochondrial biogenesis and couples red cell mass to mitochondrial mass in the heart. Circ Res. 106:1722–1730. 2010. View Article : Google Scholar : PubMed/NCBI
39	Mudalagiri NR, Mocanu MM, Di Salvo C, Kolvekar S, Hayward M, Yap J, Keogh B and Yellon DM: Erythropoietin protects the human myocardium against hypoxia/reoxygenation injury via phosphatidylinositol-3 kinase and ERK1/2 activation. Br J Pharmacol. 153:50–56. 2008. View Article : Google Scholar : PubMed/NCBI
40	Cimolai MC, Alvarez S, Bode C and Bugger H: Mitochondrial mechanisms in septic cardiomyopathy. Int J Mol Sci. 16:17763–17778. 2015. View Article : Google Scholar : PubMed/NCBI
41	Desco MC, Asensi M, Márquez R, Martínez-Valls J, Vento M, Pallardó FV, Sastre J and Viña J: Xanthine oxidase is involved in free radical production in type 1 diabetes: Protection by allopurinol. Diabetes. 51:1118–1124. 2002. View Article : Google Scholar : PubMed/NCBI
42	MacKichan ML and DeFranco AL: Role of ceramide in lipopolysaccharide (LPS)-induced signaling. LPS increases ceramide rather than acting as a structural homolog. J Biol Chem. 274:1767–1775. 1999. View Article : Google Scholar : PubMed/NCBI
43	Meng X, Ao L, Brown JM, Meldrum DR, Sheridan BC, Cain BS, Banerjee A and Harken AH: LPS induces late cardiac functional protection against ischemia independent of cardiac and circulating TNF-alpha. Am J Physiol. 273:H1894–H1902. 1997.PubMed/NCBI
44	Li L, Takemura G, Li Y, Miyata S, Esaki M, Okada H, Kanamori H, Khai NC, Maruyama R, Ogino A, et al: Preventive effect of erythropoietin on cardiac dysfunction in doxorubicin-induced cardiomyopathy. Circulation. 113:535–543. 2006. View Article : Google Scholar : PubMed/NCBI
45	Lipsic E, Westenbrink BD, van der Meer P, van der Harst P, Voors AA, van Veldhuisen DJ, Schoemaker RG and van Gilst WH: Low-dose erythropoietin improves cardiac function in experimental heart failure without increasing haematocrit. Eur J Heart Fail. 10:22–29. 2008. View Article : Google Scholar : PubMed/NCBI
46	Shravah J, Wang B, Pavlovic M, Kumar U, Chen DD, Luo H and Ansley DM: Propofol mediates signal transducer and activator of transcription 3 activation and crosstalk with phosphoinositide 3-kinase/AKT. JAKSTAT. 3:e295542014.PubMed/NCBI
47	Gross ER, Hsu AK and Gross GJ: The JAK/STAT pathway is essential for opioid-induced cardioprotection: JAK2 as a mediator of STAT3, Akt and GSK-3 beta. Am J Physiol Heart Circ Physiol. 291:H827–H834. 2006. View Article : Google Scholar : PubMed/NCBI
48	Yu H, Lee H, Herrmann A, Buettner R and Jove R: Revisiting STAT3 signalling in cancer: New and unexpected biological functions. Nat Rev Cancer. 14:736–746. 2014. View Article : Google Scholar : PubMed/NCBI
49	Gu JJ, Montealegre ZF, Robert D, Engel MS, Qiao GX and Ren D: Wing stridulation in a Jurassic katydid (Insecta, Orthoptera) produced low-pitched musical calls to attract females. Proc Natl Acad Sci USA. 109:3868–3873. 2012. View Article : Google Scholar : PubMed/NCBI
50	Asbun J, Manso AM and Villarreal FJ: Profibrotic influence of high glucose concentration on cardiac fibroblast functions: Effects of losartan and vitamin E. Am J Physiol Heart Circ Physiol. 288:H227–H234. 2005. View Article : Google Scholar : PubMed/NCBI
51	Suskin N, McKelvie RS, Burns RJ, Latini R, Pericak D, Probstfield J, Rouleau JL, Sigouin C, Solymoss CB, Tsuyuki R, et al: Glucose and insulin abnormalities relate to functional capacity in patients with congestive heart failure. Eur Heart J. 21:1368–1375. 2000. View Article : Google Scholar : PubMed/NCBI
52	Ather S, Chan W, Bozkurt B, Aguilar D, Ramasubbu K, Zachariah AA, Wehrens XH and Deswal A: Impact of noncardiac comorbidities on morbidity and mortality in a predominantly male population with heart failure and preserved versus reduced ejection fraction. J Am Coll Cardiol. 59:998–1005. 2012. View Article : Google Scholar : PubMed/NCBI
53	Paulus WJ and Tschöpe C: A novel paradigm for heart failure with preserved ejection fraction: Comorbidities drive myocardial dysfunction and remodeling through coronary microvascular endothelial inflammation. J Am Coll Cardiol. 62:263–271. 2013. View Article : Google Scholar : PubMed/NCBI
54	Ma H, Gong H, Chen Z, Liang Y, Yuan J, Zhang G, Wu J, Ye Y, Yang C, Nakai A, et al: Association of Stat3 with HSF1 plays a critical role in G-CSF-induced cardio-protection against ischemia/reperfusion injury. J Mol Cell Cardiol. 52:1282–1290. 2012. View Article : Google Scholar : PubMed/NCBI
55	Butler KL, Huffman LC, Koch SE, Hahn HS and Gwathmey JK: STAT-3 activation is necessary for ischemic preconditioning in hypertrophied myocardium. Am J Physiol Heart Circ Physiol. 291:H797–H803. 2006. View Article : Google Scholar : PubMed/NCBI
56	Roul D and Recchia FA: Metabolic alterations induce oxidative stress in diabetic and failing hearts: Different pathways, same outcome. Antioxid Redox Signal. 22:1502–1514. 2015. View Article : Google Scholar : PubMed/NCBI
57	Varga ZV, Giricz Z, Liaudet L, Hasko G, Ferdinandy P and Pacher P: Interplay of oxidative, nitrosative/nitrative stress, inflammation, cell death and autophagy in diabetic cardiomyopathy. Biochim Biophys Acta. 1852:232–242. 2015. View Article : Google Scholar : PubMed/NCBI
58	Monticone M, Taherian R, Stigliani S, Carra E, Monteghirfo S, Longo L, Daga A, Dono M, Zupo S, Giaretti W and Castagnola P: NAC, tiron and trolox impair survival of cell cultures containing glioblastoma tumorigenic initiating cells by inhibition of cell cycle progression. PLoS One. 9:e900852014. View Article : Google Scholar : PubMed/NCBI