Damage Effects Of Oxidized Soybean Oil On Primary Cultured Intestinal Epithelial Cells From Ctenopharyngodon Idellus And Its Protection

In this study, primary culture method and system evaluation index of growth ofIECs from Ctenopharyngodon idellus were established. Water-soluble-material fromoxidazied soybean oil and yeast, malondialdehyde were used in primary cultured IECsfrom Ctenopharyngodon idellus, and the adding final concentration into mediumsrespectively were111.06to888.48g/L,1.23to9.89μmol/L,10to200mg/L, as well asadding both of50to200mg/L yeast culture and4.94to9.89μmol/L MDA.The results showed that: IECs of Ctenopharyngodon idellus could be primarycultuted using mechanical scrape plus collagens digestion method and cetifugal speedson400r/min, cells grew well in M199added15%serum in6%CO2incubator, while thecell seeding concentration was2000clumps per hole. The system evaluation index ofgrowth process of primary cultured IECs were the combinations of invertedfluorescence microscope method and Giemsa staining method, MTT method, enzymeactivity of AKP and the ratio of LDH and MTT OD.Compared with the control group, added oxidized soybean oil on111.06to888.48g/L into medium inhibited cell growth and cell differentiation, and affected thecell morphology on12h, its effect results were concentration-dependent andtime-dependent. In addition, oxidized soybean oil damaged the cell structure and thecell antioxidant capacity to a certain extent. Adding888.48g/L oxidized soybean oil, cellactivity and enzyme activity of AKP decreased very significantly within12h (P <0.01),the ratio of LDH enzyme activity and MTT OD in medium increased very significantly(P<0.01), and the enzyme activity of SOD and T-AOC capacity in cultivate mediumwere decreased significantly in3to9h (P <0.01).Compared with the control group, added malondialdehyde final concentration of4.94to9.89μmol/L inhibited cell growth very significantly in3to6h, and total protein content of cells decreased very significantly within12h, the ratio of LDH enzymeactivity and MTT OD in medium increased very significantly on3h, as well as GSH-PXenzyme activity in medium on6h (P<0.01), T-AOC capacity in medium decreasedsignificantly on3h and9h (P<0.05), but SOD enzyme activity in medium changed notsignificantly in3h to12h. We observed that the effects of Malondialdehyde andwater-soluble-materials of oxidized soybean oil containing the same concentration ofMDA on IECs were similar.Compared with the control group, added final concentration of50to200mg/Lyeast culture’s water-soluble-material increased cell growth within3to6h, rate of cellproliferation was increased by42.83%. While added both the yeast culture’swater-soluble-material and MDA, the results showed that added50to200mg/L yeastculture’s water-soluble-material could protect cells from the damage of4.94to9.89μmol/L MDA. Compared with adding4.94μmol/L MDA, cell activity in groupsAdded100mg/L yeast culture’s water-soluble-material increased very significantly on6h and9h (P<0.01), and total protein content of cells increased very significantly within3to12h (P<0.01), at the same time, the ratio of LDH enzyme activity and MTT OD inmedium decreased very significantly on3h and6h(P<0.01), and GSH-PX and SODenzyme activity and T-AOC capability in medium increased to a certain extent.In addition, cells in groups adding444.24g/L water-soluble-material of oxidizedsoybean oil and4.94μmol/L malondialdehyde were balloon-like, at the same time, cellswere cellplasma vacuolation, and the nuclear of cells liked pyknosis on9h. Compared toadding444.24g/L oxidized soybean oil, the cell morphology and structure in groupsadded100mg/L yeast culture and444.24g/L of oxidized soybean oil were well.