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Tropical Journal of Pharmaceutical Research
Pharmacotherapy Group, Faculty of Pharmacy, University of Benin, Benin City, Nigeria
ISSN: 1596-5996
EISSN: 1596-5996
Vol. 16, No. 6, 2017, pp. 1259-1266
Bioline Code: pr17161
Full paper language: English
Document type: Research Article
Document available free of charge

Tropical Journal of Pharmaceutical Research, Vol. 16, No. 6, 2017, pp. 1259-1266

 en Mitochondrial transcription factor A protects human retinal endothelial cell injury induced by hypoxia
Xie, Qilian & Cao, Xu

Abstract

Purpose: To investigate the impact of mitochondrial transcription factor A (TFAM), as a modulator of NF-κB, on proliferation of hypoxia-induced human retinal endothelial cell (HREC), and the probable mechanism.
Methods: After exposure to hypoxia (1 % O2) for 5 days, cell proliferation and cell cycle of HREC were measured by MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide) assay and flow cytometry. Cell signaling and mitochondrial DNA (mtDNA) copies were determined using real-time polymerase chain reaction and Western blot. NF-κB activity was evaluated by luciferase assay.
Results: TFAM expression decreased to 40 % in HREC under hypoxic condition (p < 0.05). MTT results revealed that TFAM facilitated HREC proliferation under hypoxia (p < 0.05). Moreover, flow cytometry demonstrated that TFAM promoted HREC proliferation by accelerating cell cycle (p < 0.05). Western blot and luciferase assay exhibited NF-κB activation in HREC after TFAM overexpression (p < 0.05). Finally, real-time PCR results showed that mtDNA and targeted genes of NF-κB were upregulated 3-fold in HREC after TFAM transfection under hypoxia (p < 0.05).
Conclusion: These results indicate that NF-κB activated by TFAM protects against hypoxia-induced HREC injury by accelerating cell cycle. The ability of TFAM to enhance NF-κB signaling may be part of the mechanism of hypoxia-induced cell injury. Thus, upregulation of TFAM may help to relieve diabetic retinopathy.

Keywords
Mitochondrial transcription factor A; NF-κB; Hypoxia; Human retinal endothelial cell; Diabetic retinopathy

 
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