Effect Of PPARγ Gene On The Migration Ability Of ADCSs

Objective: Cell-directed migration is a key step in cell repair.Adipose-derived stem cells(ADSCs)have strong self-renewal and multi-directional differentiation potential.Transplantation is a potential treatment for tissue damage.Peroxisome proliferator-activated receptor γ(PPARγ)can inhibit the migration of a variety of cells.In order to verify the effect of inhibiting PPARγ gene on the migration ability of ADSCs,the PPARγ inhibitor BADGE acts on ADSCs and observes ADSCs migration and proliferation,detecting intracellular expression of matrix metalloproteinase 2(MMP2),matrix metalloproteinase 9(MMP9),and SDF/CXCR4 pathway-related proteins,and exploring the role of PPARγ on ADSCs migration,and the possible signal path.Experimental methods: First,human ADSCs were extracted for in vitro culture.Second,ADSCs were treated with BADGE containing different concentrations of PPARγinhibitors.Third,they were divided into three groups according to the purpose of the experiment: Group A: ADSCs 25μm/L BADGE common culture solution;ADSCs50μm/L BADGE common culture solution;Group C: ADSCs common culture solution;Fourth,scratch test and Transwell to observe the migration of ADSCs.PCR and Westernblot were used to detect the expression levels of MMP2,MMP9,CXCR4 genes and proteins in each group.The proliferation of ADSCs in each group was detected.Finally,the experimental data were analyzed statistically.Experimental results: 1.Migration of cells in two-dimensional space: At 72 hours,the migration rate of the ordinary culture group was about 63%,that of the 25μm/L BADGE medium group was about 43%,and that of the 50 μm/L BADGE medium group was about 24%.Compared with the liquid group,the inhibition of PPARγ gene reduced the migration capacity of ADSCs in two-dimensional space(P<0.05);2.Cell migration in three-dimensional space: 25μm/L and 50μm/L BADGE culture group ADSCs cells passing through the cell at 24 h The number was reduced by 14% and 28% respectively compared with the normal culture group;3.The expression of MMP2,MMP9,CXCR4 genes and their proteins were significantly reduced after inhibiting the PPARγ gene(P<0.05);4.The proliferation status of ADSCs in each group: after CCK-8 test,at 72 h,the number of cells in the 50μm/L BADGE culture group was reduced by 78% compared with the control group,and it was statistically significant(P<0.05);the value of 72 h cells in the 25μm/L BADGE culture group was reduced compared with the normal culture group 40%,statistically significant(P<0.05);the 72 h value in the 50μm/L BADGE medium group was reduced by 63% compared with the 25μm/L BADGE medium group,which was statistically significant(P<0.05).Conclusion: The migration ability of ADSCs decreases after the inhibition of PPARγgene;And this inhibitory effect increases with the increase of inhibitor concentration.