International Ophthalmology

, Volume 39, Issue 12, pp 2833–2842 | Cite as

Probucol promotes high glucose-induced proliferation and inhibits apoptosis by reducing reactive oxygen species generation in Müller cells

  • Xuxia Zhou
  • ShiBei Ai
  • ZhongPing ChenEmail author
  • ChenXiang Li
Original Paper



To explore the protective effect of probucol on human retinal Müller cells cultured in high glucose.


Primary Müller cells from human retinas were cultured in complete DMEM. Third-generation Müller cells were identified using glutamine synthetase (GS) antibody and randomly divided into three groups: normoglycemia (NG, 5.5 mmol/L); hyperglycemia (HG, 30 mmol/L); and hyperglycemia (30 mmol/L) with probucol (10 μmol/L; HGPB). After a 24-h intervention, cell proliferation, apoptosis, and cellular reactive oxygen species (ROS) were measured with a CCK-8 kit, flow cytometry, and DCFH-DA probe, respectively. Kelch-like ECH-associated protein 1 (Keap1), NF-E2-related factor 2 (Nrf2), and glutamate cysteine ligase catalytic subunit (GCLC) protein expression were detected by immunofluorescence staining.


For NG, HG, and HGPB, optical density (OD) values for cell proliferation were 0.98 ± 0.23, 0.58 ± 0.11, and 0.73 ± 0.11; apoptotic rates were 2.79 ± 0.52%, 7.70 ± 0.44%, and 4.00 ± 0.95%; and intracellular ROS were 20.89 ± 5.14, 55.17 ± 14.07, and 26.28 ± 4.73, respectively. Compared to NG, OD was markedly decreased (P < 0.01), apoptosis was increased (P < 0.001), and intracellular ROS level was significantly higher than in HG (P < 0.01). Compared to HG, OD was markedly increased (P < 0.01), apoptosis was meaningfully decreased (P < 0.01), and intracellular ROS level was significantly lower than in HGPB (P < 0.01). GS, Keap1, Nrf2, and GCLC had positive expression.


Probucol could inhibit intracellular ROS generation, promote proliferation, and decrease apoptosis of human retinal Müller cells cultured in high glucose. This might also be associated with Keap1/Nrf2/ARE oxidative stress signaling pathway activation.


Diabetic retinopathy Probucol Reactive oxygen species Cell proliferation Apoptosis 



We thank the staff at the Research Institute of AIER Eye for their assistance in this research and Professor Chang-Luo for his help in writing this paper.


This study was supported by the Natural Science Foundation Project of Hunan Province (No. 2018JJ2001), China, and the Research Fund Project of AIER Eye Hospital Group (No. AF1601D5), China.

Compliance with ethical standards

Conflict of interest

All authors declare that they have no conflict of interest.


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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.AIER School of OphthalmologyCentral South UniversityChangshaChina
  2. 2.Department of Fundus OculiAIER Eye Hospital of ChangshaChangshaChina
  3. 3.Department of OphthalmologyThe Seventh Affiliated Hospital of Sun Yat-sen UniversityShenzhenChina

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