Protective Effect Of Chikusetsusaponin Ⅳa On Hydrogen Peroxide Induced Injury Of Intestinal Porcine Enterocyte Cells

Chikusetsusaponin Ⅳa is a kind of saponin compound which widely existing in Panax japonicus and other Acanthopanaceae herbs,which has a variety of pharmacological activities such as hypoglycemic and hypolipidemic.IPEC-J2 cells belong to jejunal epithelial cell line of piglets,which is very important for the study of intestinal diseases of pigs.It is also widely used in the study of intestinal inflammation and permeability of pigs.Consequently,to explore the effect of natural drug molecules on hydrogen peroxide induced intestinal cell damage in pigs,provide basis for clinical prevention and treatment of animal diseases.In this study,IPEC-J2 cells were cultured in vitro to investigate the effect of H₂O₂ on IPEC-J2 at different time and concentration,and the effect of CHS-Ⅳa on IPEC-J2 under H₂O₂.In the experiment,MTT cell proliferation test was used to measure cell activity,and cell scratch test was used to study the effect of CHS-Ⅳa on the injury and repair of IPEC-J2 cells.The activities of CAT,SOD,Trx R and GSH in the supernatant of cells were detected by colorimetry,and LDH in the supernatant of cells were detected by 2,4-dinitrophenylhydrazine;Detection of MDA in cell culture supernatant by colorimetry.The relative expression of IL-6,IL-α,TNF-α,p65 and NF-κB m RNA was detected by real time PCR.The relative expression of TNF-αprotein was detected by Western blot.The results showed that the activity of IPEC-J2 cells decreased to about 50%when treated with H₂O₂ at a concentration of 1000μM for a short period of 2 hours.Low concentration of CHS-Ⅳa（50μg/ml～100μg/ml）can promote the activity of IPEC-J2 cells in a short period of time,and also promote its damage repair.CHS-Ⅳa with high concentration of 200μg/ml has obvious side effects on cells.CHS-Ⅳa of80μg/ml and 100μg/ml can regulate the redox system of IPEC-J2 cells and improve the oxidative damage caused by H₂O₂.The results showed that 80 and 100μg/ml CHS-Ⅳa could regulate the redox system of IPEC-J2 cells damaged by H₂O₂ to a certain extent,which could improve the oxidative damage caused by H₂O₂.the LDH activity in the culture supernatant of IPEC-J2 cells damaged by 1000μM H₂O₂increased significantly,but the LDH activity,MDA content decreased through CHS-Ⅳa treatment.The activities of GSH,CAT,SOD and Trx R were opposite.At the same time,after 80 and 100μg/ml CHS-Ⅳa treated IPEC-J2 cells damaged by H₂O₂,the expression of related inflammatory factors m RNA and protein decreased significantly.