Effect Of Dapagliflozin On Endoplasmic Reticulum Stress And Apoptosis In High Glucose-Stimulated Podocyte

Objective:By investigating the effects of Dapagliflozin on endoplasmic reticulu m stress and apoptosis in high glucose-stimulated podocyte(MPC-5),to provide a referen ce for the clinical application of Dapagliflozin.Method:Frozen MPC-5 in a-80℃ refrigerator was quickly thawed and cultured in RPM I 1640 medium containing 10% fetal bovine serum,5.6mmol/L D-glucose,1% Penicillin-Stre ptomycin Solution and 4ng/ml mouse gamma-interferon,and placed in the 33℃,5% CO2 cell incubator.In about 3 days,the cell cultured in RPMI 1640 medium containing 10% fetal b ovine serum,5.6 mmol/L D-glucose and 1% Penicillin-Streptomycin Solution,placed them i n the 37℃,5% CO2 cell incubator.10-14 days later when MPC-5 were differentiated and ma ture,MPC-5 were randomly assigned to the following groups to receive different treatme nts:Control group(Con),High glucose group(HG,30mmol/L,D-glucose),Mannitol group(Mn,30mmol/L D-mannitol),Dapagliflozin group(Dapa,10mmol/L Dapagliflozin),High glucose+Dapagliflozin group(HG+Dapa,30mmol/L,D-glucose+10mmol/L Dapagliflozi n),Mannitol+Dapagliflozin group(Mn,30mmol/L D-mannitol+10mmol/L Dapaglifloz in).The treatment time of each group was 48 h.The cell Viability of each group was dete rmined by the CCK-8,to make sure that the Dapagliflozin has protect effect in h igh glucose-stimulated podocyte(MPC-5)rather than in hypertonic-stimulated(by Mannit ol)podocyte(MPC-5);The apoptosis rate was detected by now cytometry;Western blotting was used to detect the expression of C/EBP homology protein(CHOP),heavy-chain binding protein(Bip),B-cell lymphoma-2(Bcl-2),BCL2-Associated X(BAX).Result:Compared with the Con group,the viability of cell in HG group and Mn group decreased,and the decreased level of the viability of cell in HG group is more serious.Compared with the Con group,the expression of CHOP and Bip increased,the expression of Bcl-2/BAX protein decreased(All P<0.05);Compared with the HG group,the viability of c ell in HG+Dapa group increased,Compared with the Mn group,the viability of cell in Mn+D apa group increased,but the increased level of the viability of cell in Mn+Dapa group i s smaller than in HG+Dapa group.Compared with the HG group,the expression of CHOP and Bip decreased,the expression of Bcl-2/BAX protein increased(All P<0.05).All have statistical significance.Conclusion:Dapagliflozin has protective effect on endoplasmic reticulum stress and apoptosis in high glucose-stimulated podocyte(MPC-5).