The Experimental Researches Of ShRNA Targeting PPARγ Gene For Prevention And Treatment Of Steroid-induced ONFH

Purpose: In this study, aiming to explore the preventive capacity of shRNA on ONFH and how the shRNA will impact on PPAR- γ.To Investigate effects of shRNA on adipogenic andosteogenic differentiation of BMSCs.Exploring shRNA directional PPARγ gene for steroid induced avascular necrosis of prevention and treatment.Methods:1 Group:The rabbit BMSCs of second filial generation were randomly divided into 6 groups, and cultivated in a concentration of 5×104 /cm2 in the six –wells plate.Normal(N): no special treatment,only normal cultivated BMSCs.Model( M): adding dexamethasone and maintaining the level of concentration at 1×10-7 mol / L.Adenoviraur(E):adding dexamethasone and adenoviral, the adenovirusDensity is 20 times thecell density.Control(I1,I2,I3): adding dexamethasoneand packed three kinds of targeting PPARγ gene adenovirus.2 Observingthe binding ofadenovirusandcells.3 Six groupsuse q PCR test mRNA expression of PPARγ and BMP-2 on the 5th,7th and10 th day.4 Automatic biochemical analyzer(ACA) were used for detecting the differences of ALP andTriglyceride content in each group.5 using Oil red O staining and cell counting methods detect adipogenic differentiation of bone marrow mesenchymal cells.Results:1 Expression of BMP-2 and PPARγ:on the 5th,7th and10 th day the expression level of BMP-2 mRNA ofgroupM and E were lower than that of group N,with statistical significa-nce(P<0.05).There was no significant difference between group N andthree groups I, the difference was not statistically significant(P >0.05).However,the expression level of PPARγ mRNA of group M and E werehigher than that of group N, with statistical significance(P<0.05),there was no significant difference between group N and three groups I,the difference was not statistically significant(P>0.05)2 Expression of triglyceride:on the 14 th day,the expression level of triglyceride of group M and E were lower than that of group N with statistical significance(P<0.05). there was no significant difference between group N and three groups I,the difference was not statistically significant(P>0.05).3 Expression of alkaline phosphatase:on the 15 th day,the expression level of alkaline phosphatase of group M and E were lower than that of group N, with statistical significance(P<0.05),there was no significant difference between group N and three groups I,the difference was not statistically significant(P>0.05).4 Expression of the number of fat cell:the number of fat cell of group M and E were more than that of group N,with statistical significance(P<0.05),there was no significant differencebetween group N and three groups I, the difference was not statistically significant(P>0.05).Conclusions:1 The shRNA can suppress the expression of the PPARγ mRNA in BMSCS, meanwhile it inhibits the differentiationofadipogenic and promotesthe differentiationof osteoblast.2 The shRNA throws new lights on prevention and treatment of steroid-induced ONFH.