Resveratrol Regulate The Extracellular Matrix Expression Via Canonical Wnt/?-catenin Pathway In Nucleus Pulposus Cells

Objective To investigate the regulatory effect of resveratrol(RES)on the extracellular matrix(ECM),and its relative molecular mechanism.Methods Ten patients receiving discectomy were collected,of which 5 patients were young with spinal burst fracture,classified as normal control group;the rest 5 patients were senile with lumbar disc herniation,classified as degenerative group.The in situ expression of ?-catenin was detected by immunohistochemistry,and the protein expressionos of collagen type II(COL II)and aggrecan were detected by western blot.The nucleus pulposus cells(NPCs)were isolated and cultured from degenerative tissues.RES treated the third-passage NPCs with or without IL-1?,to further determine the expressions of COL II and aggrecan.Moreover,NPCs treated with small interfering RNA(siRNA)Targeted silent SIRT1 or?-catenin were using Western blot,Fluorescent quantification PCR to determine the expressions of ?-catenin,SIRT1;In addition,NPCs treated with RES,IL-1?,SIRT1-siRNA were divided into control group,IL-1? group,IL-1?+RES group,SIRT1 siRNA transfection group,using the cell immunofluorescence staining to detect the nuclear translocation of beta-catenin.Finally,NPCs treated with RES,IL-1?,??catenin siRNA transfection,SIRT1 siRNA transfection were divided into Blank control group(I),IL-1? group(?),IL-1? + ?-catenin-siRNA transfection group(III),IL-1? + RES group(IV),IL-1?+ RES + SIRT1 siRNA transfection group(V),using Western blot to detect the expressions of COL II and aggrecan.Results IHC test showed that the expression of ?-catenin was significantly increased in degenerative NP tissues,meanwhile,the expressions of COL II and aggrecan were significantly decreased.In cytology experiments,results showed that after stimulation by RES,the expression of ?-catenin attenuated,and the degradation of ECM induced by IL-1? was significantly inhibited.SIRT1 expression and ?-catenin were silenced by siRNA transfection.Subsequently,after SIRT1 was silenced,Immunofluorescence results suggested that the attenuated nuclear translocation of P-catenin by RES treatment was aggravated,meanwhile,the expressions of COL II and aggrecan were decreased.In contrast,after?-catenin inhibition by siRNA,the degradation of COL ? and aggrecan by IL-1? was reversed.Conclusion RES regulated the ECM expression via Wnt/?-catenin signaling pathway,which provided a new idea for intervertebral disc degeneration disease treatment.