Di-(2-Ethylhexyl) Phthalate Induces Apoptpsis Of Gc-2spd Cell Via TR4 Pathway

Objective This study investigated the role of the TR4 pathway in apoptosis induced by DEHP,which resulted in reproductive damage,and revealed the DEHP induced possible mechanisms of sperm cells apoptosis.Methods GC-2 spd cells were exposed to various DEHP concentrations(0,50,100,or200μmol/L)1.Reverse Transcription Polymerase Chain Reaction(RT-PCR)was used to analyze the levels of Bcl-2,Bax,caspase 9,caspase 3,Cyt-C m RNA.2.Western Blot was used to analyze the levels of TR4,Bcl-2,Bax,procaspase 9,procaspase 3,Cyt-C protein expression 3.Flow cytometry instrument detecting GC-2spd cell apoptosis.Results1.With the increase of the concentrations of DEHP,the survival rate of cell decreased gradually,high dose group was obviously higher than that in control group(P < 0.05),but low dose group compared with control group the difference is not obvious.2.The exposed group Bax,Cyt-C caspase-9 and caspase-3 m RNA expression levels were elevated.The high dose exposure group of Bax and caspase-9 and,high dose exposure group of Cyt-C,the middle dose group of caspase-3 compared with the control group,the difference has statistical significance(P < 0.05),bcl-2 m RNA expression levels were decreased,and all exposed groups of Bcl-2 compared with the control group,the difference has statistical significance(P < 0.05).3.The exposed group TR4,Bcl-2,procaspase-3,procaspase-9 protein expression levels decreased,Bax and Cyt-C protein expression level increased,which Bcl-2 in high dose group,high dose group of procaspase-3,middle dose of procaspase-9 and procaspase-3 group,high dose of procaspase-9 and procaspase-3group and high dose group of Bax,high dose group of Cyt-C compared with control group difference has statistical significance(P < 0.05).ConclusionDEHP can regulat TR4 on mitochondria,by reducing the Bcl-2 protein expression,increase the expression of cytochrome C protein,activated caspase-9caspase-3,at last cause cells apoptosis.