Caspase3 activity during the progress of HUVEC apoptosis induced by homocysteine / 中国病理生理杂志

ABSTRACT

AIM: In this research, the apoptosis mechanism in human umbilical vein endothelial cells (HUVEC) exposed to increasingly high Hcy concentrations was investigated by examining the intracellular signaling pathways. METHODS: HUVEC were cultured and pretreated with Hcy. Twenty-four h after Hcy treatment, cell apoptosis was detected by Annexin V and DNA ladder analysis. Caspase3 and c-IAP1/2 mRNA expression were examined using reverse transcription-polymerase chain reaction (RT-PCR) and protein expression detected by Western blotting. The activation of caspase 3 was measured with the specific substrate DEVD-AMC. RESULTS: Hcy (0.3 mmol/L) induced apoptosis of HUVEC. Caspase 3 mRNA and protein expressions increased and activation enhanced with the concentration of increasing Hcy, but c-IAP2 mRNA and protein expression decreased. CONCLUSION: Hcy induces apoptosis of HUVEC in vivo by activating caspase 3 and inhibiting protein c-IAP-2 mRNA and protein expression.