Ozone alleviates ischemia/reperfusion injury by inhibiting mitochondrion‐mediated apoptosis pathway in SH‐SY5Y cells

Abstract

Cerebral ischemia/reperfusion (I/R) injuries are common and often cause severe complications. Ozone has been applied for protecting I/R injury in animal models of several organs including cerebra, but the detailed mechanism remains unclear. 3‐(4,5‐Dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) assay and lactate dehydrogenase measurement were used to determine the influence of ozone on cell activity and damage of SH‐SY5Y cells. Some redox items such as catalase (CAT), malondialdehyde (MDA), glutathione peroxidase (GSH‐Px), and superoxide dismutase (SOD) were measured by enzyme‐linked immunosorbent assay. The mitochondrial membrane potential (ΔΨm) was determined by JC‐1 assay. Cytochrome‐c (cyt‐c) level in the cytoplasm and mitochondrion was measured by western blotting. Apoptosis was determined by flow cytometry, and some apoptosis‐related molecules were detected by quantitative real‐time polymerase chain reaction and western blotting. Ozone alleviated oxidative damage by increasing GSH‐Px, SOD, CAT, and decreasing MDA. Ozone decreased mitochondrial damage caused by I/R injury and inhibited the release of cyt‐c from mitochondrion to cytoplasm in SH‐SY5Y cells. The cell apoptosis caused by I/R was inhibited by ozone, and ozone could decrease apoptosis by increasing the ratio of Bcl‐2/Bax and inhibiting caspase signaling pathway in SH‐SY5Y cells. Ozone has the ability of maintaining redox homeostasis, decreasing mitochondrion damage, and inhibiting neurocytes apoptosis induced by I/R. Therefore, ozone may be a promising protective strategy against cerebral I/R injury.

RETRACTED ARTICLE: Restored microRNA-326-5p Inhibits Neuronal Apoptosis and Attenuates Mitochondrial Damage via Suppressing STAT3 in Cerebral Ischemia/Reperfusion Injury

“…CI/RI activates neuronal apoptosis and leads to hippocampal and cortical damage [3]. At present, the most common clinical application is thrombolytic for CI/RI [4]. However, reperfusion would increase the production of reactive oxygen species (ROS), leading to intracellular DNA damage, oxidative stress-related damage, protein oxidation and lipid peroxidation, thus further deteriorating bloodbrain barrier and edema [5].…”

Ozone induces tolerance against cardiomyocytes oxygen‑glucose deprivation/reperfusion through inhibition of autophagy pathway

“…The ratio of Bcl-2/BAX is the key factor that decides whether apoptosis happens to cells following injuries. In line with the results of a previous study (25), the present study demonstrated that incubation with a low concentration of ozone (20 µg/ml) significantly upregulated the ratio of Bcl-2/BAX and downregulated the expression of cleaved caspase-3 when compared with high concentration (60 µg/ml) ozone and the OGD/R group. Additionally, the results of flow cytometry further demonstrated that incubation with low concentration ozone significantly attenuated the apoptosis rate when compared with the OGD/R group.…”

A Flavonoid-Rich Extract of Mandarin Juice Counteracts 6-OHDA-Induced Oxidative Stress in SH-SY5Y Cells and Modulates Parkinson-Related Genes

“…SH-SY5Y cells were plated at density of 5 × 10 5 cells/well in 6-well plates and were treated with MJe 24 h later at a concentration 0.1 and 0.5 mg/mL for 1 h. Then, 50 µM of 6-OHDA was added for additional 24 h. Then, the assays were conducted according to the manufacturer’s protocols on cell lysates. The absorbance was recorded by an iMark™ microplate reader (Bio-Rad Laboratories) at 450 nm for SOD, 570 nm for CAT, and 405 nm for GSH [ 25 , 26 ].…”