Protective Effects Of Clostridium Butyricum On Intestinal Epithelial Cells Against Oxidative Stress

Oxidative stress has the characteristics of high incidence,strong concealment and great harmfulness in the gastrointestinal tract of humans and animals.It is easy to pose a threat to human health and cause huge losses to the development of animal husbandry.Clostridium butyricum is a strictly anaerobic spore-producing probiotic bacteria existing in human and animal intestines.It has many excellent characteristics such as regulating intestinal microecological balance,enhancing immune function,anti-oxidation and strong tolerance,and has good application prospects.a Clostridium butyricum CB-M1 with potential beneficial effects was isolated from the intestinal tract of healthy piglets.The bacteria were isolated by anaerobic culture.The target strain was analyzed by morphological identification,physiological and biochemical methods and 16 S r DNA gene sequence homology analysis,and a phylogenetic tree was constructed.Staphylococcus aureus and Enterococcus faecium were selected to analysis the protective effect of C.butyricum CB-M1 on intestinal epithelial cell line Caco-2.The concentrations of glyoxal(GO),methylglyoxal(MGO),4-hydroxyhexenal(HHE)and 4?Hydroxynonenal(HNE)were 400 ?mol/L,400 ?mol/L,60 ?mol/L and 60 ?mol/L,respectively;multiplicity of infection(MOI)is 1:1(Number of bacteria: number of cells).The cell viability test showed that within 30 h of co-culture C.butyricum CB-M1 did not significantly promote the growth of Caco-2 cells,and S.aureus could significantly promote the growth of Caco-2 cells(p<0.05),while E.faecium could significantly reduce the cell viability of Caco-2 cells(p<0.01).Combined with the results of ATRA inhibition of Nrf2-ARE pathway,we found that C.butyricum CB-M1 did not induce a significant increase in reactive oxygen species(ROS)levels in Caco-2 cells,while S.aureus and E.faecium could induce a significant increase in ROS levels in Caco-2 cells.C.butyricum CB-M1 could significantly alleviate the toxicity and oxidative stress of GO,MGO,HHE and HNE to Caco-2 cells,and induce Caco-2 cells to express IL-10 to protect cell growth.S.aureus could aggravate the toxicity and oxidative stress of GO,MGO,HHE and HNE to Caco-2 cells,and induce the expression of IL-6 in Caco-2 cells and damage the growth of cells.E.faecium showed no significant effect on the toxicity and oxidative stress of GO,MGO,HHE and HNE to Caco-2 cells.In summary,this study successfully isolated and identified a strain of C.butyricum CB-M1 from healthy piglets,which does not induce elevated ROS levels in Caco-2 cells to cause oxidative stress,but could induce Caco-2 cells to express anti-inflammatory cytokine IL-10 and significantly alleviate the induction of oxidative stress in Caco-2 cells by GO,MGO,HHE and HNE to protect cells.It laid a foundation for further application of C.butyricum CB-M1 in microecological preparation.