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Dissociation of HKII in retinal epithelial cells induces oxidative stress injury in the retina.

International journal of molecular medicine (2019-08-23)
Liqun Chu, Lin Xiao, Bing Xu, Jingmei Xu
ZUSAMMENFASSUNG

The retina is sensitive to injury resulting from oxidative stress (OS) due to its high oxygen consumption. Patients with retinitis pigmentosa suffer from excessive OS. N‑acetylcysteine (NAC) is used as a mucolytic agent for the clinical treatment of disorders, such as chronic bronchitis and other pulmonary diseases. The aim of the present study was to investigate the role of hexokinase 2 (HKII) in retinal OS injury. Amyloid β (Aβ)1‑40 was used to establish a cellular model of OS. Cell viability was measured with a Cell Counting Kit‑8 assay, and the apoptosis, reactive oxygen species (ROS) and mitochondrial membrane potential (MMP) of cells were analyzed via flow cytometry with corresponding kits. The mRNA and protein levels were detected by reverse transcription‑quantitative PCR and western blot analyses, respectively. It was observed that Aβ1‑40 reduced the expression of HKII in the mitochondria of retinal pigment epithelial ARPE cells and impaired mitochondrial antioxidant functions. Additionally, knockdown of HKII promoted apoptosis, and increased ROS levels and the MMP. NAC attenuated the inhibition of mitochondrial functions induced by Aβ1‑40. The knockdown of HKII was revealed to decrease the levels of Bcl‑2, manganese superoxide dismutase (SOD) and copper‑zinc‑SOD, and increase the levels of cleaved caspase‑3, Bax and cytochrome c. The present findings suggested that the dissociation of HKII induced by OS induces apoptosis and mitochondrial damage. This study provided improved understanding of the mechanisms underlying the effects of OS on retinal epithelial cells.

MATERIALIEN
Produktnummer
Marke
Produktbeschreibung

Sigma-Aldrich
Fluorometric Intracellular Ros Kit, sufficient for 200 fluorometric tests (green)
Sigma-Aldrich
(Tyr[SO3H]27)Cholecystokinin-Fragment 26-33 Amid, ≥97% (HPLC), powder
Sigma-Aldrich
2X Cell Lysis Buffer (Item J)