Effect Of BHB On Proliferation And PLD1 Expression Of Human Astroglioma U251 Cells

Objective:The effects of different concentrations of ?-hydroxybutyrate(BHB)on the proliferation of human astroglioma U251 cells and the expression of phospholipase D1(PLD1)in vitro were studied.Methods:1.The classical malignant cell line U251 cells in human astrocytes were used as experimental subjects,and the ketogenic microenvironment was constructed by BHB.The effect of BHB on the proliferation of human astrocyte U251 cells and the effect of BHB on the expression of phospholipase D1(PLD1)were investigated by using in vitro experimental systems.2.Experimental group: divided into control group and experimental group.The control group was only cultured in normal complete medium without any additional drugs.The experimental group was divided into four different concentrations of BHB treatment groups(2,4,8,10mmol/L),which was equivalent to adding four different concentrations of BHB treatment cells(2,4,8,10mmol/L)into the normal complete medium.3.The effects of different concentrations of BHB on the proliferation of glioma U251 cells were detected by MTT assay at 24 hours and 48 hours after drug administration.The effect ofdifferent concentrations of BHB on the expression of PLD1 protein in glioma U251 cells was detected by Western blot.Results:1.The MTT results show:(1)U251 cells were treated with2,4,8,10 mmol/L BHB for 24 hours,and the cell inhibition rates were(14.26±6.35)%,(19.8±7.31)%,(22.19±5.79)%,(25.07±5.32)%,respectively.Compared with the control group,the viability of cells in 2mmol/L group,4mmol/L group,8mmol/L group and 10mmol/L group decreased(p<0.05),and the cell viability decreased with the increase of drug concentration.(2)U251 cells were treated with 2,4,8,10 mmol/L BHB for 48 hours,the cell inhibition rates were(3.79±2.32)%,(6.72±1.99)%,(10.71±1.89)%,(17.06±2.54)%,respectively.Compared with the control group,the viability of cells in 2mmol/L group,4mmol/L group,8mmol/L group and 10mmol/L group decreased(p<0.05),and the cell viability decreased with the increase of drug concentration.(3)Comparison of the same concentration of BHB on U251 cells in vitro for24 hours and 48 hours,and the cell inhibition in the 24 hours group was more obvious,which was statistically significant(p<0.05),suggesting that BHB had an inhibitory effect on the proliferation of U251 cells.It was concentration dependent but not time dependent.2.The Western blot results show:(1)U251 cells were treated with 2,4,8,10 mmol/L BHB for 24 hours,and the relative expression levels of PLD1 were0.694±0.003,0.681±0.004,0.652±0.006and0.634±0.005,respectively.Com-pared with the control group,the expression of PLD1 in the 2mmol/L group,4mmol/L group,8mmol/L group and 10mmol/L group decreased(p<0.05),and the expression of PLD1 decreased with the increase of the concentration.(2)U251 cells were treated with 2,4,8,10 mmol/L BHB for 48 hours,and the relative expression levels of PLD1 were1.321±0.002,1.242±0.006,1.221±0.005and0.933±0.005,respectively.Com-pared with the control group,the expression of PLD1 in the 2mmol/L group,4mmol/L group,8mmol/L group and 10mmol/L group decreased(p<0.05),and the expression of PLD1 decreased with the increase of the concentration.(3)Comparison of the same concentration of BHB on U251 cells in vitro for 24 hours and 48 hours,and the expression of PLD1 in the 24 hours group was lower,which was statistically significant(p<0.05),suggesting that BHB has an inhibitory effect on PLD1 expression.It is concentration dependent but not time dependent.Conclusions: 1.The BHB can inhibit the proliferation of glioma U251 and down-regulate the expression of PLD1 in a concentrationdependent but not time-dependent manner.2.The BHB can inhibit the proliferation of U251 cells,which may be related to the down-regulation of PLD1 expression.