Overexpression of steroid receptor coactivators alleviates hyperglycemia-induced endothelial cell injury in rats through activating the PI3K/Akt pathway

Abstract

Hyperglycemia is a major factor in vascular endothelial injury that finally leads to a cardiovascular event. Steroid receptor coactivators (SRCs) are a group of non-DNA binding proteins that induce structural changes in steroid receptors (nuclear receptors) critical for transcriptional activation. SRCs, namely, SRC-1, SRC-2, and SRC-3, are implicated in the regulation of vascular homeostasis. In this study we investigate the role of SRCs in hyperglycemia-induced endothelial injury. Aortic endothelial cells were prepared from normal and diabetic rats, respectively. Diabetic rats were prepared by injection of streptozotocin (50 mg/kg, i.p.). The expression levels of SRC-1 and SRC-3 were significantly decreased in endothelial cells from the diabetic rats. Similar phenomenon was also observed in aortic endothelial cells from the normal rats treated with a high glucose (25 mM) for 4 h or 8 h. The expression levels of SRC-2 were little affected by hyperglycemia. Overexpression of SRC-1 and SRC-3 in high glucose-treated endothelial cells significantly increased the cell viability, suspended cell senescence, and inhibited cell apoptosis compared with the control cells. We further showed that overexpression of SRC-1 and SRC-3 markedly suppressed endothelial injury through restoring nitric oxide production, upregulating the expression of antioxidant enzymes (SOD, GPX, and CAT), and activating the PI3K/Akt pathway. The beneficial effects of SRC-1 and SRC-3 overexpression were blocked by treatment with the PI3K inhibitor LY294002 (10 mM) or with the Akt inhibitor MK-2206 (100 nM). In conclusion, hyperglycemia decreased SRC-1 and SRC-3 expression levels in rat aortic endothelial cells. SRC-1 and SRC-3 overexpression might protect against endothelial injury via inhibition of oxidative stress and activation of PI3K/Akt pathway.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

References

  1. 1.

    Yang L, Shao J, Bian Y, Wu H, Shi L, Zeng L, et al. Prevalence of type 2 diabetes mellitus among inland residents in China (2000-2014): a meta-analysis. J Diabetes Investig. 2016;7:845–52.

  2. 2.

    Kozakova M, Palombo C. Diabetes mellitus, arterial wall, and cardiovascular risk assessment. Int J Environ Res Public Health. 2016;13:201.

  3. 3.

    Rajendran P, Rengarajan T, Thangavel J, Nishigaki Y, Sakthisekaran D, Sethi G, et al. The vascular endothelium and human diseases. Int J Biol Sci. 2013;9:1057–69.

  4. 4.

    Hoffman RP. Vascular endothelial dysfunction and nutritional compounds in early type 1 diabetes. Curr Diabetes Rev. 2014;10:201–7.

  5. 5.

    Paneni F, Beckman JA, Creager MA, Cosentino F. Diabetes and vascular disease: pathophysiology, clinical consequences, and medical therapy: part I. Eur Heart J. 2013;34:2436–43.

  6. 6.

    Du XL, Edelstein D, Dimmeler S, Ju Q, Sui C, Brownlee M. Hyperglycemia inhibits endothelial nitric oxide synthase activity by posttranslational modification at the Akt site. J Clin Invest. 2001;108:1341–8.

  7. 7.

    Arunachalam G, Samuel SM, Marei I, Ding H, Triggle CR. Metformin modulates hyperglycaemia-induced endothelial senescence and apoptosis through SIRT1. Br J Pharmacol. 2014;171:523–35.

  8. 8.

    Wang S, Peng Q, Zhang J, Liu L. Na+/H+exchanger is required for hyperglycaemia-induced endothelial dysfunction via calcium-dependent calpain. Cardiovasc Res. 2008;80:255–62.

  9. 9.

    Stashi E, York B, O’Malley BW. Steroid receptor coactivators: servants and masters for control of systems metabolism. Trends Endocrinol Metab. 2014;25:337–47.

  10. 10.

    Johnson AB, O’Malley BW. Steroid receptor coactivators 1, 2, and 3: critical regulators of nuclear receptor activity and steroid receptor modulator (SRM)-based cancer therapy. Mol Cell Endocrinol. 2012;348:430–9.

  11. 11.

    York B, O’Malley BW. Steroid receptor coactivator (SRC) family: masters of systems biology. J Biol Chem. 2010;285:38743–50.

  12. 12.

    Leo C, Chen JD. The SRC family of nuclear receptor coactivators. Gene. 2000;245:1–11.

  13. 13.

    Walsh CA, Qin L, Tien JC, Young LS, Xu J. The function of steroid receptor coactivator-1 in normal tissues and cancer. Int J Biol Sci. 2012;8:470–85.

  14. 14.

    Yuan Y, Xu J. Loss-of-function deletion of the steroid receptor coactivator-1 gene in mice reduces estrogen effect on the vascular injury response. Arterioscler Thromb Vasc Biol. 2007;27:1521–7.

  15. 15.

    Ying H, Willingham MC, Cheng SY. The steroid receptor coactivator-3 is a tumor promoter in a mouse model of thyroid cancer. Oncogene. 2008;27:823–30.

  16. 16.

    Fleet T, Zhang B, Lin F, Zhu B, Dasgupta S, Stashi E, et al. SRC-2 orchestrates polygenic inputs for fine-tuning glucose homeostasis. Proc Natl Acad Sci USA. 2015;112:E6068–6077.

  17. 17.

    Stashi E, Lanz RB, Mao J, Michailidis G, Zhu B, Kettner NM, et al. SRC-2 is an essential coactivator for orchestrating metabolism and circadian rhythm. Cell Rep. 2014;6:633–45.

  18. 18.

    Reineke EL, York B, Stashi E, Chen X, Tsimelzon A, Xu J, et al. SRC-2 coactivator deficiency decreases functional reserve in response to pressure overload of mouse heart. PLoS ONE. 2012;7:e53395.

  19. 19.

    Sherr CJ, Roberts JM. Living with or without cyclins and cyclin-dependent kinases. Genes Dev. 2004;18:2699–711.

  20. 20.

    Nie XQ, Chen HH, Zhang JY, Zhang YJ, Yang JW, Pan HJ, et al. Rutaecarpine ameliorates hyperlipidemia and hyperglycemia in fat-fed, streptozotocin-treated rats via regulating the IRS-1/PI3K/Akt and AMPK/ACC2 signaling pathways. Acta Pharmacol Sin. 2016;37:483–96.

  21. 21.

    McGuire PG, Orkin RW. Isolation of rat aortic endothelial cells by primary explant techniques and their phenotypic modulation by defined substrata. Lab Invest. 1987;57:94–105.

  22. 22.

    Chen X, Qin L, Liu Z, Liao L, Martin JF, Xu J. Knockout of SRC-1 and SRC-3 in mice decreases cardiomyocyte proliferation and causes a noncompaction cardiomyopathy phenotype. Int J Biol Sci. 2015;11:1056–72.

  23. 23.

    Gonzalez-Arenas A, Hansberg-Pastor V, Hernandez-Hernandez OT, Gonzalez-Garcia TK, Henderson-Villalpando J, Lemus-Hernandez D, et al. Estradiol increases cell growth in human astrocytoma cell lines through ERalpha activation and its interaction with SRC-1 and SRC-3 coactivators. Biochim Biophys Acta. 2012;1823:379–86.

  24. 24.

    Fiorentino TV, Prioletta A, Zuo P, Folli F. Hyperglycemia-induced oxidative stress and its role in diabetes mellitus related cardiovascular diseases. Curr Pharm Des. 2013;19:5695–703.

  25. 25.

    Chen M, Wang W, Ma J, Ye P, Wang K. High glucose induces mitochondrial dysfunction and apoptosis in human retinal pigment epithelium cells via promoting SOCS1 and Fas/FasL signaling. Cytokine. 2016;78:94–102.

  26. 26.

    Dassanayaka S, Readnower RD, Salabei JK, Long BW, Aird AL, Zheng YT, et al. High glucose induces mitochondrial dysfunction independently of protein O-GlcNAcylation. Biochem J. 2015;467:115–26.

  27. 27.

    Wu N, Shen H, Liu H, Wang Y, Bai Y, Han P. Acute blood glucose fluctuation enhances rat aorta endothelial cell apoptosis, oxidative stress and pro-inflammatory cytokine expression in vivo. Cardiovasc Diabetol. 2016;15:109.

  28. 28.

    Cifarelli V, Lee S, Kim DH, Zhang T, Kamagate A, Slusher S, et al. FOXO1 mediates the autocrine effect of endothelin-1 on endothelial cell survival. Mol Endocrinol. 2012;26:1213–24.

  29. 29.

    Cvoro A, Tzagarakis-Foster C, Tatomer D, Paruthiyil S, Fox MS, Leitman DC. Distinct roles of unliganded and liganded estrogen receptors in transcriptional repression. Mol Cell. 2006;21:555–64.

  30. 30.

    Hinton AO Jr, Yang Y, Quick AP, Xu P, Reddy CL, Yan X, et al. SRC-1 regulates blood pressure and aortic stiffness in female mice. PLoS ONE. 2016;11:e0168644.

  31. 31.

    Sahar S, Reddy MA, Wong C, Meng L, Wang M, Natarajan R. Cooperation of SRC-1 and p300 with NF-kappaB and CREB in angiotensin II-induced IL-6 expression in vascular smooth muscle cells. Arterioscler Thromb Vasc Biol. 2007;27:1528–34.

  32. 32.

    Yuan Y, Liao L, Tulis DA, Xu J. Steroid receptor coactivator-3 is required for inhibition of neointima formation by estrogen. Circulation. 2002;105:2653–9.

  33. 33.

    Wang W, Bian K, Vallabhaneni S, Zhang B, Wu RC, O’Malley BW, et al. ERK3 promotes endothelial cell functions by upregulating SRC-3/SP1-mediated VEGFR2 expression. J Cell Physiol. 2014;229:1529–37.

  34. 34.

    Carrero P, Okamoto K, Coumailleau P, O’Brien S, Tanaka H, Poellinger L. Redox-regulated recruitment of the transcriptional coactivators CREB-binding protein and SRC-1 to hypoxia-inducible factor 1alpha. Mol Cell Biol. 2000;20:402–15.

  35. 35.

    Zeng L, Xiao Q, Chen M, Margariti A, Martin D, Ivetic A, et al. Vascular endothelial cell growth-activated XBP1 splicing in endothelial cells is crucial for angiogenesis. Circulation. 2013;127:1712–22.

  36. 36.

    Li J, Xiong J, Yang B, Zhou Q, Wu Y, Luo H, et al. Endothelial cell apoptosis induces TGF-beta signaling-dependent host endothelial-mesenchymal transition to promote transplant arteriosclerosis. Am J Transplant. 2015;15:3095–111.

  37. 37.

    Karbach S, Jansen T, Horke S, Heeren T, Scholz A, Coldewey M, et al. Hyperglycemia and oxidative stress in cultured endothelial cells--a comparison of primary endothelial cells with an immortalized endothelial cell line. J Diabetes Complicat. 2012;26:155–62.

  38. 38.

    Brouwers O, Niessen PM, Haenen G, Miyata T, Brownlee M, Stehouwer CD, et al. Hyperglycaemia-induced impairment of endothelium-dependent vasorelaxation in rat mesenteric arteries is mediated by intracellular methylglyoxal levels in a pathway dependent on oxidative stress. Diabetologia. 2010;53:989–1000.

  39. 39.

    Xi G, Shen X, Wai C, Vilas CK, Clemmons DR. Hyperglycemia stimulates p62/PKCzeta interaction, which mediates NF-kappaB activation, increased Nox4 expression, and inflammatory cytokine activation in vascular smooth muscle. FASEB J. 2015;29:4772–82.

  40. 40.

    Wang W, Wang WH, Azadzoi KM, Dai P, Wang Q, Sun JB, et al. Alu RNA accumulation in hyperglycemia augments oxidative stress and impairs eNOS and SOD2 expression in endothelial cells. Mol Cell Endocrinol. 2016;426:91–100.

Download references

Acknowledgements

Financial support was provided by the Key Program for Social Development, Science and Technology of Shaanxi Province, China (2016SF-077), and the Social Development Guidance Program-Medical Research Project of Xi’an, China (SF1509).

Author contributions

XJQ and CLL designed research and wrote the paper; XJQ, CLL, MZS, LZ, and XLL performed research; and XJQ, CLL, LZ, and XLL analyzed data.

Author information

Correspondence to Xiao-juan Quan.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Quan, X., Liang, C., Sun, M. et al. Overexpression of steroid receptor coactivators alleviates hyperglycemia-induced endothelial cell injury in rats through activating the PI3K/Akt pathway. Acta Pharmacol Sin 40, 648–657 (2019). https://doi.org/10.1038/s41401-018-0109-4

Download citation

Keywords

  • diabetics
  • streptozotocin
  • hyperglycemia
  • endothelial cells
  • steroid receptor coactivators
  • cell senescence
  • cell apoptosis
  • oxidative stress
  • PI3K/Akt pathway
  • LY294002
  • MK-2206