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Updated: Jan 1, 2026

Malachite Green Assay for the Discovery of Heat-Shock Protein 90 Inhibitors
Published on: January 20, 2023
[Mechanisms of heat shock proteins inhibiting C2C12 cell apoptosis induced by hydrogen peroxide]
Wei-Min Xiao1, Bi-Mei Jiang, Yong-Zhong Shi
1Department of Pathophysiology, Xiangya School of Medicine, Central South University, Changsha 410078, China.
Objective:
To explore the mechanisms of C2C12 cell apoptosis induced by hydrogen peroxide (H2O2) which is inhibited by heat shock proteins.
Methods:
The expression of heat shock proteins in mouse embryonic myogenic cell line, C2C12 cell,was induced by heat shock response. C2C12 cell apoptosis induced by 0.5 mmol/L hydrogen peroxide (H2O2) was determined by Hoechst 33258 staining. The activities of caspase -3,8,9 were assayed by caspase colorimetric assay kit and Western-blotting. The release of cytochrome C from mitochondria was observed by Western-blotting of cell mitochondria and cytosol fractions.
Results:
The expression of HSP70 and alphaB-crystallin in C2C12 cell significantly increased at 24 hour after the heat shock response. Heat shock response could inhibit the release of cytochrome c from mitochondria to cytoplasm,the activation of caspase -3,8,9 and the subsequent apoptosis induced by H2O2 in C2C12 cell.
Conclusion:
HSPs can inhibit the C2C12 cell apoptosis through interference with the activation of both mitochondrial and death receptor pathways, which can provide new clues for the prevention of cardiovascular diseases.
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