Ms. Caiping Zhuang Profile

Caiping Zhuang

at First Affiliated Hospital Of Jinan Univ

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Proceedings Article | 14 February 2012 Paper

Hydrogen peroxide induces apoptosis via a mitochondrial pathway in chondrocytes

Cai-ping Zhuang, Qian Liang, Xiao-ping Wang, Tong-sheng Chen

Proceedings Volume 8225, 82251Q (2012) https://doi.org/10.1117/12.905786

KEYWORDS: Cell death, Luminescence, Hydrogen, Cartilage, Flow cytometry, Confocal microscopy, Life sciences, Molecular mechanisms, Rhodamine, Carbon dioxide

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The degenerative joint disease such as osteoarthritis (OA) is closely associated with the death of chondrocytes in apoptosis fashion. Hydrogen peroxide (H₂O₂), higher expression following acute damage in OA patients, has been shown to be up-regulated during apoptosis in a bulk of experimental models. This study was aimed to explore the mechanism of H2O2-induced rabbit chondrocytes apoptosis. Articular cartilage was biopsied from the joints of 6 weeks old New Zealand rabbits. Cell Counting Kit (CCK-8) assay was used to assess the inhibitory effect of H₂O₂ on cell viability. H₂O₂ treatment induced a remarkable reduction of cell viability. We used flow cytometry to assess the form of cell death with Annexin-V/PI double staining, and found that H2O2 treatment induced apoptosis in a dose-and time-dependent manner. Exposure of chondrocytes to 1.5 mM of H₂O₂ for 2 h induced a burst apoptosis that can be alleviated by N-acetyl cysteine (NAC) pretreatment, an anti-oxidant amino-acid derivative. Loss of mitochondria membrane potential (▵Ψm) was evaluated using confocal microscopy imaging and flow cytometry (FCM). H₂O₂ treatment induced a marked reduction of ▵Ψm, and the abrupt disappearance of ▵Ψm occurred within 5 minutes. These results indicate that H₂O₂ induces a rapid apoptosis via a mitochondrial pathway in rabbit chondrocytes.

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