3-Monochloropropane-1,2-diol induced cell apoptosis in HK2 cells via mitochondria-dependent pathway
[Objective]This study investigated the toxic effects of the food contaminant 3-monochloropropane-1,2-diol(3-MCPD)on the human renal tubular epithelial cell line(HK2)and examined the underlying mechanisms.[Methods]Cell viability,morphology,mitochondrial function,enzyme activities,and apoptosis-associated proteins expressions were assessed using MTT assay,Hoechst 33342 staining,biochemical test kits and Western blot,respectively.[Results]Results showed a concentration-dependent decrease in cell viability when 3-MCPD was below 80 mmol/L for 24 h.Treatment with 20 and 30 mmol/L 3-MCPD over varying durations also led to a time-dependent reduction in cell viability.Additionally,3-MCPD at 20,40 and 60 mmol/L significantly increased LDH release,decrease mitochondrial membrane potential,and lowered ATP levels.Fluorescence microscopy revealed notable morphological changes,including bright blue fluorescence,nuclear shrinkage and apoptosis bodies.Furthermore,3-MCPD treatment downregulated the anti-apoptotic protein Bcl2,upregulated the pro-apoptotic protein Bax,and increased Caspase 9 and Caspase 3 activity in a dose-dependent manner.[Conclusion]3-MCPD exhibits toxic effects on HK2 cells and induces apoptosis through a mitochondria-dependent pathway.
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