Effects Of Vitamin E Levels On Oxidative Stress In Dairy Cow Mammary Epithelial Cells

With the in-depth study of oxidative stress in humans,more and more experiments have pointed out that vitamin E,as an antioxidant,has an antioxidant effect at low doses,and vitamin E will change to a pro-oxidative direction as the dose increases.Therefore,this experiment took dairy cow mammary epithelial cells(MAC-T)as the experimental object to explore the effects of different doses of vitamin E and H2O2 on it,and to provide theoretical reference for the rational use of vitamin E in production practice.This test is mainly divided into two parts.The MTT colorimetric method is used to detect the cell viability,the oxidation indicators TAS,SOD,CAT,MDA,TOS and ALP are detected by Elisa kit,and the cell permeability is detected by transepithelial resistance meter.The TEER value of the treated cells indicates that the tight junction protein was detected using real-time PCR.Experiment 1: After the cell viability was detected by MTT colorimetry,the concentration and time of the treatment group were set as 400 μmol/L/10 h,600 μmol/L/6h,900 μmol/L/4h,and the concentration of the treatment group was 10 μmol/L and 20 μmol/L.,40μmol/L vitamin E relieved for 2,4,6,and 8 hours,and the data of cell viability and oxidation indexes were analyzed.In the second part,cells were treated with vitamin E at concentrations of 300 μmol/L,600 μmol/L,900μmol/L,1200 μmol/L,1500 μmol/L,1800 μmol/L for 2,4,6,and 8 hours,respectively,and then based on experiment one,experiment one was used.The hydrogen peroxide treatment group in the third group was combined with the vitamin E treatment group in the second experiment to explore the effect of the collision between the two stressors,and to detect the cell viability,oxidation indexes and cell membrane permeability.The results of the first part of the previous experiments showed that the concentration of 20μmol/L vitamin E effectively reduced the content of MDA and TOS in cells,restored the cell viability and the production of SOD and CAT in cells,and the oxidative stress caused by different concentrations and time required The remission time of vitamin E is different,the oxidative stress caused by high concentration of hydrogen peroxide requires longer time to relieve vitamin E,and vice versa,the time required to relieve low-level damage is shorter.High doses of vitamin E can cause oxidative damage to cells,and the contents of MDA and ALP in cells are significantly increased,which inhibits the production of SOD and CAT.The increase of intracellular ALP causes the TEER value to drop sharply,and the cell membrane is damaged,causing cell death..It can be seen from the experimental results of the cell survival rate and oxidation index of the combination of the two stressors that the combination of the two stressors is extremely significant(p<0.01)lower than the control group regardless of the order of addition,and higher than the damage to cells caused by the addition of the two stressors alone..All in all,different degrees of oxidative damage require different vitamin E mitigation times,and high doses of vitamin E can cause oxidative damage to cells,which will continue over time,and the combination of the two stressors will affect cells.cause more damage.