Effects of high glucose on store operated calcium entry and related protein expressions in H9C2 and neonatal rat ventricular cardiomyocytes

doi:10.3969/j.issn.1006-7795.2020.03.017

Abstract

Abstract: Objective To investigate the effects of high glucose on store operated calcium entry (SOCE) and the endogenous stromal interaction molecule 1 (STIM1) expression and its activated structure "puncta" in rat embryonic cardiomyocytes line (H9C2) and neonatal rat ventricular myocytes (NRVMs), which may be involved in the pathogenic mechanism of diabetic cardiomyopathy (DCM). Methods H9C2 and NRVMs cardiomyocytes were divided into two groups:normal glucose group (5.5 mmol/L) and high glucose group (25 mmol/L) treated and cultured for 48 h. The alteration of Ca²⁺ influx mediated by SOCE pathway were observed by laser confocal microscope after treated with thapsigargin (TG, 2 μmol/L) in H9C2 and NRVMs cells. The expression levels of STIM1 and calcium release-activated calcium channel modulator 1 (Orai1) were detected by Western blotting. The aggregated formation of STIM1 puncta in NRVMs was detected by chemical crosslinking and immuno-fluorescence staining. Direct interaction between endogenous STIM1 and Orai1 in NRVMs was observed by co-immunoprecipitation. Results Compared with control group, TG stimulated sarcoplasmic reticulum(SR) Ca²⁺ store depletion and induced more SOCE (P<0.05), and increased STIM1 and Orai1 expressions (P<0.01) in the H9C2 and NRVMs cells cultured with high glucose. While the interaction between endogenous STIM1/Orai1 was not altered between the two groups of cells, the aggregated formation of STIM1 puncta was significantly increased after SR Ca²⁺ store depletion by TG in NRVMs cells cultured with high glucose (P<0.01). Conclusion High glucose could up-regulate the expression levels of STIM1 and Orai1 proteins and promote the formation of STIM1 puncta, leading to an increased SOCE in H9C2 and NRVMs cardiomyocytes, which implicated a potential mechanism involved in diabetic cardiomyopathy.

Key words: high glucose, store operated calcium entry (SOCE), stromal interaction molecule 1 (STIM1), calcium release-activated calcium channel modulator 1 (Orai1)

CLC Number:

Shaletanati·Talabieke, Sun Zhipeng, Wang Luqi, You Hongjie, Luo Dali. Effects of high glucose on store operated calcium entry and related protein expressions in H9C2 and neonatal rat ventricular cardiomyocytes[J]. Journal of Capital Medical University, 2020, 41(3): 411-420.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2020.03.017

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2020/V41/I3/411

References

[1] Tate M, Grieve D J, Ritchie R H, et al. Are targeted therapies for diabetic cardiomyopathy on the horizon?[J]. Clin Sci, 2017, 131(10):897-915.
[2] De Jong A M, Maass A H, Oberdorf-Maass S U, et al. Mechanisms of atrial structural changes caused by stretch occurring before and during early atrial fibrillation[J]. Cardiovasc Res, 2011, 89(4):754-765.
[3] Petersen C E,Wolf M J,Smyth J T. Drosophila suppression of store-operated calcium entry causes dilated cardiomyopathy of the heart[J].Biol Open, 2020,9(3):bio049999.
[4] Bénard L, Oh J G, Cacheux M, et al. Cardiac STIM1 silencing impairs adaptive hypertrophy and promotes heart failure through inactivation of mTORC2/Akt signaling[J]. Circulation, 2016, 133(15):1458-1471.
[5] Shaw R M, Colecraft H M. L-type calcium channel targeting and local signaling in cardiac myocytes[J]. Cardiovasc Res, 2013, 98(2):177-186.
[6] Elaib Z, Saller F, Bobe R. The calcium entry-calcium refilling coupling[J]. Adv Exp Med Biol, 2016, 898:333-352.
[7] Shen W W, Frieden M, Demaurex N, et al. Local cytosolic Ca²⁺ elevations are required for stromal interaction molecule 1(STIM1) de-oligomerization and termination of store-operated Ca²⁺ entry[J]. J Biol Chem, 2011, 286(42):36448-36459.
[8] Amcheslavsky A, Wood M L, Yeormin A V, et al. Molecular biophysics of Orai store-operated Ca²⁺ channels[J]. Biophys J, 2015, 108(2):237-246.
[9] Stathopulos P B, Schindl R, Fahrner M, et al. STIM1/Orai1 coiled-coil interplay in the regulation of store-operated calcium entry[J]. Nat Commun, 2013, 4:2963.
[10] Zhou Y, Srinivasan P, Razavi S, et al. Initial activation of STIM1, the regulator of store-operated calcium entry[J]. Nat Struct Mol Biol, 2013, 20(8):973-981.
[11] Deler I, Plenk P, Fahrner M, et al. The extended transmembrane Orai1 N-terminal (ETON) region combines binding interface and gate for Orai1 activation by STIM1[J]. J Biol Chem, 2013, 288(40):29025-29034.
[12] Thompson J L, Shuttleworth T J. A plasma membrane-targeted cytosolic domain of STIM1 selectively activates ARC channels, an arachidonate-regulated store-independent Orai channel[J]. Channels (Austin), 2012, 6(5):370-378.
[13] Ohba T, Watanabe H, Murakami M, et al. Essential role of STIM1 in the development of cardiomyocyte hypertrophy[J]. Biochem Biophys Res Commun, 2009, 389(1):172-176.
[14] Stathopulos P B, Zheng L, Li G Y, et al. Structural and mechanistic insights into STIM1-mediated initiation of store-operated calcium entry[J]. Cell, 2008, 135(1):110-122.
[15] Hirve N, Rajanikanth V, Hogan P G, et al. Coiled-coil formation conveys a STIM1 signal from ER lumen to cytoplasm[J]. Cell Rep, 2018, 22(1):72-83.
[16] Yang X, Jin H, Cai X, et al. Structural and mechanistic insights into the activation of stromal interaction molecule 1(STIM1)[J]. Proc Natl Acad Sci U S A, 2012, 109(15):5657-5662.
[17] López E, Salido G M, Rosado J A, et al. Unraveling STIM2 function[J]. J Physiol Biochem, 2012, 68(4):619-633.
[18] Bonhenry D, Schober R, Schmidt T, et al. Mechanistic insights into the Orai channel by molecular dynamics simulations[J]. Semin Cell Dev Biol, 2019, 94:50-58.
[19] Madl J, Weghuber J, Fritsch R, et al. Resting state Orai1 diffuses as homotetramer in the plasma membrane of live mammalian cells[J]. J Biol Chem, 2010, 285(52):41135-41142.
[20] Hou X, Pedil, Diver M M, et al. Crystal structure of the calcium release-activated calcium channel Orai[J]. Science, 2012, 338(6112):1308-1313.
[21] Perni S, Dynes J L, Yeromin A V, et al. Nanoscale patterning of STIM1 and Orai1 during store-operated Ca²⁺ entry[J]. Proc Natl Acad Sci U S A, 2015, 112(40):5533-5542.
[22] Palty R, Isacoff E Y, et al. Cooperative binding of stromal interaction molecule 1(STIM1) to the N and C Termini of calcium release-activated calcium modulator 1(Orai1)[J]. J Biol Chem, 2016, 291(1):334-341.
[23] Tirado-Lee L, Yamashita M, Prakriya M. Conformational changes in the Orai1 C-terminus evoked by STIM1 binding[J]. Plos One, 2015, 10(6):e0128622.
[24] Maléth J, Choi S, Muallem S, et al. Translocation between PI(4,5)P2-poor and PI(4,5)P2-rich microdomains during store depletion determines STIM1 conformation and Orai1 gating[J]. Nat Commun, 2014, 5:5843.
[25] Li X, Wu G, Yang Y, et al. Calmodulin dissociates the STIM1-Orai1 complex and STIM1 oligomers[J]. Nat Commun, 2017, 8(1):1042.
[26] Wu M M, Covington E D, Lewis R S, et al. Single-molecule analysis of diffusion and trapping of STIM1 and Orai1 at endoplasmic reticulum-plasma membrane junctions[J]. Mol Biol Cell, 2014, 25(22):3672-3685.
[27] Luo X, Hojayev B, Jiang N, et al. STIM1-dependent store-operated Ca²⁺ entry is required for pathological cardiac hypertrophy[J]. J Mol Cell Cardiol, 2012, 52(1):136-147.
[28] Eder P. Cardiac remodeling and disease:SOCE and TRPC Signaling in cardiac pathology[J]. Adv Exp Med Biol, 2017, 993:505-521.
[29] Golovina V A. Visualization of localized store-operated calcium entry in mouse astrocytes. Close proximity to the endoplasmic reticulum[J]. J Physiol, 2005, 564:737-749.
[30] Zhang B, Jiang J, Yue Z, et al. Store-operated Ca²⁺ entry (SOCE) contributes to angiotensin II-induced cardiac fibrosis in cardiac fibroblasts[J]. J Pharmacol Sci, 2016, 132(3):171-180.
[31] Hunton D L, Lucchesi P A, Pang Y, et al. Capacitative calcium entry contributes to nuclear factor of activated T-cells nuclear translocation and hypertrophy in cardiomyocytes[J]. J Biol Chem, 2002, 277(16):14266-14273..
[32] Voelkers M, Salz M, Herzog N, et al. Orai1 and STIM1 regulate normal and hypertrophic growth in cardiomyocytes[J]. J Molecular Cell Cardiol, 2010, 48(6):1329-1334.
[33] Ohba T, Watanabe H, Murakami M, et al. Essential role of STIM1 in the development of cardiomyocyte hypertrophy[J]. Biochem Biophys Res Commun, 2009, 389(1):172-176.
[34] Kumar S, Kain V, Sitasawad S L, et al. High glucose-induced Ca²⁺ overload and oxidative stress contribute to apoptosis of cardiac cells through mitochondrial dependent and independent pathways[J]. Biochim Biophys Acta, 2012, 1820(7):907-920.
[35] He F, Wu Q, Xu B, et al. Suppression of STIM1 reduced intracellular calcium concentration and attenuated hypoxia/reoxygenation induced apoptosis in H9C2 cells[J]. Biosci Rep, 2017, 37(6):BSR20171249.
[36] Pang Y, Hunton D L, Bounelis P, et al. Hyperglycemia inhibits capacitative calcium entry and hypertrophy in neonatal cardiomyocytes[J]. Diabetes, 2002, 51(12):3461-3467.
[37] Correll R N, Goonasekera S A, Van Berlo J H, et al. STIM1 elevation in the heart results in aberrant Ca²⁺ handling and cardiomyopathy[J]. J Mol Cell Cardiol, 2015, 87:38-47.
[38] Bartoli F, Bailey M A, Rode B, et al. Orai1 channel inhibition preserves left ventricular systolic function and normal ca handling after pressure overload[J]. Circulation, 2020, 141(3):199-216.
[39] Sorrentino A, Borghetti G, Zhou Y, et al. Hyperglycemia induces defective Ca²⁺ homeostasis in cardiomyocytes[J]. Am J Physiol Heart Circ Physiol, 2017, 312(1):150-161.
[40] Cesario D A, Brar R, Shivkumar K. Alterations in ion channel physiology in diabetic cardiomyopathy[J]. Endocrinol Metab Clin North Am,2006,35(3):601-610.
[41] Zhou H, Yue Y, Wang J, et al. Melatonin therapy for diabetic cardiomyopathy:A mechanism involving Syk-mitochondrial complex I-SERCA pathway[J]. Cell Signal, 2018, 47:88-100.
[42] Gui L, Zhu J, Lu X, et al. S-Nitrosylation of STIM1 by neuronal nitric oxide synthase inhibits store-operated Ca²⁺ entry[J]. J Mol Biol, 2018, 430(12):1773-1785.
[43] Zhu-Mauldin X, Marsh S A, Zou L, et al. Modification of STIM1 by O-linked N-acetylglucosamine (O-GlcNAc) attenuates store-operated calcium entry in neonatal cardiomyocytes[J]. J Biol Chem, 2012, 287(46):39094-39106.