The Effects Of Gliclazide On Proliferation, Differentiation And Apoptosis In Mouse Osteoblastic-like Cells MC3T3-E1

Object:To investigate the effects of gliclazide on proliferation, differentiation and apoptosis of MC3T3-E1 in normal glucose. Method:MC3T3-E1 cells are cultured and divided into control group, gliclazide administration group with different gliclazide concentrations(10,20 and 100μM, respectively)exposing for 48 hours.We use CCK-8 assay to detect the proliferation rate.Quantitative flow cytometry is used to detected the cell apoptosis.The m RNA of collagen I(COL-1),osteopontin(OPN) and osteogenic factor runt related gene 2(Runx2) are detected using semi-quantitative RT-PCR.The expression of apoptosisassociation genes Bcl-2 and Bax are detected by Western blot. Results:(1)Compared with control group, different gliclazide concentrations stimulate osteoblast proliferation, the best concentration of gliclazide is 20μM.10μM gliclazide group,20μM gliclazide group and 100μM gliclazide group comparing with the control group,the proliferation rates increase by 16.6%,20.2%,13.7%(P <0.05).(2) The early apoptotic rate of the control group, 10μM gliclazide group,20μM gliclazide group and 100μM gliclazide group are 8.83%,5.47%,2.87%,7.33%. Compared with control group, different gliclazide concentrations decrease the early apoptotic rate(P <0.05), the best concentration of gliclazide is 20 u M.(3)Gliclazide stimulates COL-I,OPN and Runx2 m RNA expression(P<0.05). The most best concentration of gliclazide is 20μM. The expression of COL-1 m RNA and OPN m RNA have a significant positive correlation with Runx2 binding activity(r=0.899, P<0.01; r=0.854, P<0.01, respectively).(4) The value of Bcl-2/β-actin in the control group, 10μM gliclazide group,20μM gliclazide group and 100μM gliclazide group are orderly 1.22±0.025,1.48±0.035,1.81±0.066,1.30±0.015;the value of Bax/β-actin are accordingly 0.83±0.046,0.62±0.015 0.53±0.036 0.72±0.025.Compared with control group, different gliclazide concentrations increase the expression of Bcl-2 ptotein and decrease the expression of Bax ptotein(P<0.05),the best concentration of gliclazide is 20μM.The ratio of Bcl-2/Bax have a significant negative correlation with the early apoptotic rate(r=-0.935, P<0.01) Conclusion:Gliclazide improves the proliferation, differentiation and inhibit apoptosis of MC3T3-E1 in normal glucose.Gliclazide plays a protective role on osteoblasts.