Inhibition of endothelin A receptor protects brain microvascular endothelial cells against hypoxia‑induced injury

  • Authors:
    • Xi Liu
    • Fen Deng
    • Zhen Yu
    • Yunlan Xie
    • Changlin Hu
    • Lifen Chen
  • View Affiliations

  • Published online on: April 16, 2014     https://doi.org/10.3892/ijmm.2014.1744
  • Pages: 313-320
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Abstract

Endothelin-1 (ET-1)-induced cell damage is commonly involved in ischemia/hypoxia-associated diseases. PD155080 [sodium 2-benzo (1.3)dioxol-5-yl-3-benzyl-4-(4‑metho­xyphenyl)-4-oxobut-2-enoate] is a selective endothelin A receptor (ETAR) antagonist that inhibits ET-1‑induced cell damage. The aim of this study was to investigate the effects of PD155080 on hypoxia-induced rat brain microvascular endothelial cell (BMEC) injury. BMECs were isolated from the cerebral cortex of Wistar rats and cultured in an anoxia chamber, containing 95% N2 and 5% CO2 for 12 h. BMEC injury was assessed by determining cellular ultra-microstructural changes and cell viability by MTT assay, trypan blue (TB) staining and measuring the lactate dehydrogenase (LDH) levels. ET-1 mRNA expression was detected by in situ hybridization and reverse transcription PCR (RT-PCR); the ET-1 protein level was measured by radioimmunoassay. Following exposure to hypoxic conditions, the viability of the BMECs was markedly decreased and the ultrastructure of the BMECs was damaged, as demonstrated by chromatin margination, chromatin agglutination, plasma edema, the increased number of intracellular liposomes and vacuoles, mitochondrial swelling and the expansion of a rough surfaced endoplasmic reticulum. The levels of ET-1 and ET-1 mRNA expression in the BMECs were increased following exposure to hypoxic conditions. Of note, the administration of PD155080 greatly enhanced the viability of the BMECs and ameliorated hypoxia-induced cellular injury. PD155080 also inhibited hypoxia-induced ET-1 production by the BMECs. In conclusion, PD155080 exerts protective effects against hypoxia-induced BMEC injury.

References

1 

Shostak HDC, Lemasters JJ, Edgell CJ, et al: Role of ICE-like proteases in endothelial cell hypoxic and reperfusion injury. Biochem Biophys Res Commun. 231:844–847. 1997. View Article : Google Scholar : PubMed/NCBI

2 

Luo J, Martinez J, Yin X, et al: Hypoxia induces angiogenic factors in brain microvascular endothelial cells. Microvasc Res. 83:138–145. 2012. View Article : Google Scholar : PubMed/NCBI

3 

Faller DV: Endothelial cell responses to hypoxic stress. Clin Exp Pharmacol Physiol. 26:74–84. 1999. View Article : Google Scholar

4 

Lo AC, Chen AY, Hung VK, et al: Endothelin-1 overexpression leads to further water accumulation and brain edema after middle cerebral artery occlusion via aquaporin 4 expression in astrocytic end-feet. J Cereb Blood Flow Metab. 25:998–1011. 2005. View Article : Google Scholar

5 

Kourembanas S, Marsden PA, Mcquillan LP, et al: Hypoxia induced endothelin gene expression and secretion in cultured human endothelium. J Clin Invest. 88:1054–1060. 1991. View Article : Google Scholar : PubMed/NCBI

6 

Fernandez N, Monge L, Garcia-Villalon AL, et al: Endothelin-1-induced in vitro cerebral venoconstriction is mediated by endothelin ETA receptors. Eur-J-Pharmacol. 294:483–490. 1995. View Article : Google Scholar : PubMed/NCBI

7 

Moldes O, Sobrino T, Blanco M, et al: Neuroprotection afforded by antagonists of endothelin-1 receptors in experimental stroke. Neuropharmacology. 63:1279–1285. 2012. View Article : Google Scholar : PubMed/NCBI

8 

Maguire JJ, Kuc RE, Doherty AM, et al: Potency of 155080, an orally active ETA receptor antagonist, determined for human endothelin receptors. J Cardiovasc Pharmacol. 26(suppl 3): S362–S364. 1995. View Article : Google Scholar : PubMed/NCBI

9 

Nagy Z, Vastag M, Kolev K, et al: Human cerebral microvessel endothelial cell culture as a model system to study the blood-brain interface in ischemic/hypoxic conditions. Cell Mol Neurobiol. 25:201–210. 2005. View Article : Google Scholar : PubMed/NCBI

10 

Nagy Z, Vastag M, Skopal J, et al: Human brain microvessel endothelial cell culture as a model system to study vascular factors of ischemic brain. Keio J Med. 45:200–206. 1996. View Article : Google Scholar : PubMed/NCBI

11 

Wenbin Wu, Changlin HU and Weixue TANG: Microvascular endothelial cell culture of Wistar rat cerebral cortex. Journal of Chongqing Medical University. 27:151–152. 2002.

12 

Martinez-Orgado J, Gonzalez R, Alonso MJ, et al: Endothelial factors and autoregulation during pressure changes in isolated newborn piglet cerebral arteries. Pediatr-Res. 44:161–167. 1998. View Article : Google Scholar : PubMed/NCBI

13 

Korzeniewski C and Callewaert DM: An enzyme-release assay for natural cytotoxicity. J Immunol Methods. 64:313–320. 1983. View Article : Google Scholar : PubMed/NCBI

14 

Patel TR, Galbraith S, McAuley MA and McCulloch J: Endothelin-mediated vascular tone following focal cerebral ischaemia in the cat. J Cereb Blood Flow Metab. 16:679–687. 1996. View Article : Google Scholar : PubMed/NCBI

15 

Goodwin AT, Smolenski RT, Gray CC, Jayakumar J, Amrani M and Yacoub MH: Role of endogenous endothelin on coronary reflow after cardioplegic arrest. J Thorac Cardiovasc Surg. 122:1167–1173. 2001. View Article : Google Scholar : PubMed/NCBI

16 

Chen Y, McCarron RM, Golech S, et al: ET-1- and NO-mediated signal transduction pathway in human brain capillary endothelial cells. Am J Physiol Cell Physiol. 284:C243–C249. 2003. View Article : Google Scholar : PubMed/NCBI

17 

Chang CZ, Winardi D, Lin CL, et al: Attenuation of hemolysate-induced cerebrovascular endothelial cell injury and of production of endothelin-1 and big endothelin-1 by an endothelin-converting enzyme inhibitor. Surg Neurol. 58:181–187. 2002. View Article : Google Scholar

18 

Schaller BJ: The role of endothelin in stroke: experimental data and underlying pathophysiology. Arch Med Sci. 2:1462006.

19 

Ergul A: Endothelin-1 and diabetic complications: focus on the vasculature. Pharmacol Res. 63:477–482. 2011. View Article : Google Scholar : PubMed/NCBI

20 

Yamashita K, Discher DJ, Hu J, Bishopric NH and Webster KA: Molecular regulation of the endothelin-1 gene by hypoxia. Contributions of hypoxia-inducible factor-1, activator protein-1, GATA-2, AND p300/CBP. J Biol Chem. 276:12645–12653. 2001. View Article : Google Scholar : PubMed/NCBI

21 

Moreau P, d’Uscio LV, Shaw S, Takase H, Barton M and Lüscher TF: Angiotensin II increases tissue endothelin and induces vascular hypertrophy: reversal by ET(A)-receptor antagonist. Circulation. 96:1593–1597. 1997. View Article : Google Scholar : PubMed/NCBI

22 

Barton M, d’Uscio LV, Shaw S, Meyer P, Moreau P and Lüscher TF: ET(A) receptor blockade prevents increased tissue endothelin-1, vascular hypertrophy, and endothelial dysfunction in salt-sensitive hypertension. Hypertension. 31:499–504. 1998. View Article : Google Scholar

23 

Lüscher Thomas F and Barton Matthias: Endothelins and endothelin receptor antagonists: therapeutic considerations for a novel class of cardiovascular drugs. Circulation. 102:2434–2440. 2000.PubMed/NCBI

24 

Terese PR and Nilsson GE: Endothelin induced cerebral vasoconstriction in rainbow trout, detected in a novel in vitro preparation. Neurosci Lett. 325:195–198. 2002. View Article : Google Scholar : PubMed/NCBI

25 

Kaundal RK, Deshpande TA, Gulati A, et al: Targeting endothelin receptors for pharmacotherapy of ischemic stroke: current scenario and future perspectives. Drug Discov Today. 17:793–804. 2012. View Article : Google Scholar : PubMed/NCBI

26 

Takasu A, Matsushima S, Takino M, et al: Effect of an endothelin-1 antagonist, BQ-485, on cerebral oxygen metabolism after complete global cerebral ischemia in dogs. Resuscitation. 34:65–69. 1997. View Article : Google Scholar : PubMed/NCBI

27 

Leung JW, Chung SS and Chung SK: Endothelial endothelin-1 over-expression using receptor tyrosine kinase tie-1 promoter leads to more severe vascular permeability and blood brain barrier breakdown after transient middle cerebral artery occlusion. Brain Res. 1266:121–129. 2009. View Article : Google Scholar

28 

Legos JJ, Lenhard SC, Haimbach RE, et al: selective ET(A) receptor antagonism: perfusion/diffusion MRI used to define treatable stroke model, time to treatment and mechanism of protection. Exp Neurol. 212:53–62. 2008. View Article : Google Scholar : PubMed/NCBI

29 

Khatibi NH, Lee LK, Zhou Y, et al: Endothelin receptor-A (ETa) inhibition fails to improve neonatal hypoxic-ischemic brain injury in rats. Acta Neurochir Suppl. 111:207–212. 2011. View Article : Google Scholar : PubMed/NCBI

30 

Danielyan L, Mueller L, Proksch B, et al: Similar protective effects of BQ-123 and erythropoietin on survival of neural cells and generation of neurons upon hypoxic injury. Eur J Cell Biol. 84:907–913. 2005. View Article : Google Scholar : PubMed/NCBI

31 

Doherty AM, Patt WC, Repine J, et al: Structure-activity relationships of a novel series of orally active nonpeptide ETA and ETA/B endothelin receptor-selective antagonists. J Cardiovasc Pharmacol. 26(suppl 3): S358–S361. 1995. View Article : Google Scholar

32 

Friedrich B, Gerald W and Stephen H: Defective intracellular calcium handling in monocrotaline-induced right ventricular hypertrophy: protective effect of long-term endothelin-A receptor blockade with 2-benzo[1,3]dioxol-5-yl-3-benzyl-4-(4-methoxy-phenyl-)-4-oxobut-2-enoate-sodium (PD 155080). J Pharmacol Exp Ther. 300:442–449. 2002.PubMed/NCBI

33 

Spiers JP, Kelso EJ, McDermott BJ, Scholfield CN and Silke B: Endothelin-1 mediated inhibition of the acetylcholine-activated potassium current from rabbit isolated atrial cardiomyocytes. Br J Pharmacol. 119:1427–1437. 1996. View Article : Google Scholar

34 

Wanecek M, Oldner A, Sundin P, Alving K, Weitzberg E and Rudehill A: Effects on haemodynamics by selective endothelin ET(B) receptor and combined endothelin ET(A)/ET(B) receptor antagonism during endotoxin shock. Eur J Pharmacol. 386:235–245. 1999. View Article : Google Scholar : PubMed/NCBI

35 

Potter GS, Johnson RJ and Fink GD: Role of endothelin in hypertension of experimental chronic renal failure. Hypertension. 30:1578–1584. 1997. View Article : Google Scholar : PubMed/NCBI

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July 2014
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APA
Liu, X., Deng, F., Yu, Z., Xie, Y., Hu, C., & Chen, L. (2014). Inhibition of endothelin A receptor protects brain microvascular endothelial cells against hypoxia‑induced injury. International Journal of Molecular Medicine, 34, 313-320. https://doi.org/10.3892/ijmm.2014.1744
MLA
Liu, X., Deng, F., Yu, Z., Xie, Y., Hu, C., Chen, L."Inhibition of endothelin A receptor protects brain microvascular endothelial cells against hypoxia‑induced injury". International Journal of Molecular Medicine 34.1 (2014): 313-320.
Chicago
Liu, X., Deng, F., Yu, Z., Xie, Y., Hu, C., Chen, L."Inhibition of endothelin A receptor protects brain microvascular endothelial cells against hypoxia‑induced injury". International Journal of Molecular Medicine 34, no. 1 (2014): 313-320. https://doi.org/10.3892/ijmm.2014.1744