Effect Of Inhibitor Of Differentiation 1 Gene Transfection On Bone Morphogenetic Protein 2 Promoting Chondrogenic Gene Expressions Of Rabbit Intervertebral Cartilage Endplate Cells

Objective: To demonstrate the effect of inhibitor of differentiation 1(Id1) gene transfection on bone morphogenetic protein 2(BMP-2) promoting the expressions of collagen type II and aggrecan in intervertebral cartilage endplate cells(EPCs).Methods:(1) EPCs were harvested from the New Zealand white rabbits, the 2nd generation EPCs were used for experiment.(2)EPCS was transfected GFP LV, Id1 over expressed LV, RNAi Id1 LV. The morphology of EPCs were observed by fluorescence microscope. The m RNA express change of Id1 in EPCs was depicted by fold change though RT-PCR. The synthesis of Id1 protein was tested by Western blot.(3)EPCS were tranfected by BMP-2 and GFP LV,BMP-2 LV and Id1 over expressed LV, BMP-2 LV and RNAi Id1 LV. EPCs tranfected by BMP-2 LV was set as control group. ACAN and COL II m RNA were tested via RT-PCR. ELISA was used to test the change of COL II and ACAN protein.Results:(1)No significant morphology change happened to EPCs after tranfection by Lentiviral. The EPCs were effectively transfected by the high expression Id1 lentivirus and RNAi Id1 lentivirus; Id1 gene m RNA in EPCs was also dramatically interfered.(2)The expressions of COL II and ACAN m RNA and synthesis of COL II and ACAN protein were significantly higher in BMP-2 lentivirus and high expression Id1 LV groups than control group(P<0.05).(3)The expression of COL II and ACAN protein were down regulated in the cartilage endplate cells while the expression of Id1 was declined(P<0.05).Conclusion: Up-regulation of Id1 gene could enhance the effects of BMP-2 on the synthesis of COL II and ACAN in EPCs.