Inhibition of caspase‐3‐like activity prevents apoptosis while retaining functionality of human chondrocytes in vitro

摘要: Apoptosis was induced in a human chondrocyte cell line, T/C 28a4, by treatment with various stimuli, including camptothecin, tumor necrosis factor-α, staurosporine, okadaic acid, and reduced serum conditions. All stimuli cytosolic DEVDase activity, coincident apoptosis. Caspase activities the lysates were characterized quantitated peptide cleavage profiles. To confirm that results not related to immortalized nature of primary chondrocytes also shown undergo apoptosis under similar conditions, which resulted increased activity. There little or no caspase-1 (interleukin-1β-converting enzyme) caspase-8-like activity apoptotic cells. In all cases, irreversible nonselective caspase inhibitor, Z-VAD-FMK, caspase-3-selective Ac-DMQD-CHO, inhibited apoptosis, whereas caspase-1-selective Ac-YVAD-CHO, had effect. Human stably transiently transfected type-II collagen gene (COL2A1) regulatory sequence driving luciferase reporter as specific marker expression. Treatment cells camptothecin factor-α plus cycloheximide significantly COL2A1 transcriptional Significantly, cotreatment Z-VAD-FMK Ac-DMQD-CHO maintained COL2A1-reporter indicating prevention caspase-3 inhibition sufficient maintain functionality assessed retention promoter