Effects Of Glucose Concentration On The Arginine Absorption And Transportation In Inestinal Epithelial Cells Of Piglets

To investigate the effects of glucose concentration on the uptake and transport in intestinal epithelial cells,the piglet intestinal epithelial cells(IPEC-J2)was used to be a model in vitro.The cells were starved in DMEM(without amino)containing EBSS balanced salt solution and vitamin for 2h when the cells reached 90% confluence.To gain the 15 kinds of culture medium mixed with glucose and arginine,the 0mM/L,0.7mM/L,1.4mM/L,2.1mM/L,4.2mM/L arginine were added into three mediums containing 25mM/L,75mM/L and 225mM/L glucose,respectively.Then the cells were treated with 15 mediums,respectively.At 20,40,80 and 160 mins after treatment,the proliferations of(IPEC-J2)cells were detected.In addition,the mRNA and protein levels of absorption and transporters of the arginine were detected in this study.The results as follows:1 Effects of arginine and glucose on cell proliferation was detected.The results showed that the cell proliferation was significantly increased after added the arginine and glucose.The main effect difference is very significant(P<0.01).A tendency to increase first and then is decrease of cell proliferation with the increase of arginine concentration.And the cell proliferation was more significant after treated with the 1.4mM/L and 2.1mM/L arginine(P<0.05).Similarly,with the increase of glucose concentration,a tendency to increase first and then decrease of cell proliferation was observed.The cell proliferation was significantly increased after treated with 75mM/L glucose(P<0.05).2 Effects of arginine and glucose on mRNA expression of b0,+AT and y+LAT1 were detected.The results showed that the concentrations of glucose and arginine Significantly affect the mRNA expressions of transporters,including b0,+AT and y+LAT1(P<0.05).And a Significant interaction effects were observed in mRNA expressions of b0,+AT and y+LAT1(P<0.01).At 40 mins after treatment,75mM/L glucose and 2.1mM/L arginine significantly increased the mRNA expression of b0,+AT(P<0.05).However,at 80 mins after treatment,75mM/L glucose and 2.1mM/L arginine significantly increased the mRNA expression y+LAT1 mRNA(P<0.05).3 Effects of arginine and glucose on protein levels of b0,+AT and y+LAT1 were detected.The optimum concentration and response time of arginine and glucose were used according to the results of experiment 2.The western blot results showed that the protein expression of b0,+AT and y+LAT1 was consistent with the mRNA results.At 40 mins after treatment,2.1mM/L arginine significantly increased the b0,+AT protein levels compared with other groups(P<0.05).At 80 mins after treatment,2.1mM/L arginine significantly increased the b0,+AT protein levels compared with other groups(P<0.05).In conclusion,the concentrations of glucose and arginine had significant effects on the proliferation of piglet intestinal epithelial cells,the expression of absorption and transportation carrier gene,and a significant interaction effects were observed in mRNA expression of b0,+AT and y+LAT1.The absorption,transporter gene expression and protein synthesis of piglet intestinal epithelial cells proliferation was significantly increased after treated with 75mM/L glucose and 2.1mM/L arginine...