Study On The Protective Effect Of Ellagic Acid In IPEC-J2 Cells

Objective:The aim of this study was to observe the effects of ellagic acid(EA)on the proliferation,tight junction,cell cycle and apoptosis of IPEC-J2 cells,and to investigate the possible mechanism of action which will provide an experimental basis for the development and application of ellagic acid.Methods:The influence of EA on IPEC-J2 cell proliferation was determined by MTT assay.The effects of ellagic acid on the expression of tight junction related genes Zo-1,Claudin and Occludin was detected by RT-PCR in IPEC-J2 cells.RT-PCR and flow cytometry were uses to determinate the effects of IPEC-J2 cells on the expression of cell cycle related genes cyclinA,cyclinB,cyclinC,cyclinD,cyclinE and cell cycle.The effects of IPEC-J2 cells on apoptosis related genes caspase-3,caspase-8,caspase-9,Bax,bcl-2,and apoptosis rate were determined by the method of the RT-PCR;flow cytometry and Elisa.Results:(1)Ellagic acid can increase cell proliferation,meanwhile,can improve transcription level of tight junction related genes Zo-1,Claudin and Occluding,which showed that ellagic acid regulates IPEC-J2 cells tight junction with Zo-1,Claudin and Occludin proteins.It was indicated that ellagic acid can protect cell membrane barrier function and improve cell survival rate.(2)Ellagic acid can improve the transcription level of CyclinA,CyclinB,CyclinC,CyclinD and CyclinE,and decrease the percentage of cells in phase S and G2/M.Which indicated that ellagic acid regulates IPEC-J2 cells cycle with CyclinA、CyclinB、CyclinC、CyclinD、CyclinE and promote the synthesis of DNA and reduce the proportion of cells in phase S and G2/M,thereby promoting the renewal of cell cycle and promoting the proliferation of IPEC-J2 cells.(3)Ellagic acid can decrease the transcription level of Bax,Caspase-3,Caspase-8,Caspase-9 which indicated that ellagic acid may inhibit the expression of Bax and Caspase-9,increase the ratio of Bcl-2/Bax in mitochondrial pathway.Meanwhile,it reduced the expression of caspse-8 in death receptor pathway then reduced the expression of caspase-3 to inhibit apoptosis.It was suggested that ellagic acid may regulate the mitochondrial pathway and death receptor pathway of apoptosis,and may be the first to induce the inhibitory effect by activating the death receptor pathway to realize the role of inhibiting cell apoptosis.Conclusion:Ellagic acid inhibits apoptosis through mitochondrial pathway and death receptor pathway.It can accelerate the synthesis and repair of DNA and promote the expression of cell cycle relate genes and shortening cell cycle by promoting the expression of closely linked genes,maintaining the integrity of the intestinal mucosal barrier and improving the permeability of IPEC-J2 cells so as to achieve the proliferation and protection of the cells.