Effect of LncRNA NEAT1 on osteogenic differentiation of human bone marrow mesenchymal stem cells through miR-195-5p/TXNIP axis
Objective:To investigate the effect of LncRNA NEAT1 on osteogenic differentiation of human bone marrow mesenchymal stem cells(HBMSCs)through the miR-195-5p/TXNIP axis.Methods:The changes in expression of NEAT1,TXNIP mRNA,and miR-195-5p in HBMSCs before and af-ter osteogenic differentiation were observed.According to the treatment,HBMSCs were assigned in-to silence NEAT1(si NEAT1)group,negative control(si NC)group,miR-195-5p mimics group,negative control(mimics NC)group,si NEAT1+negative control(inhibitor NC)group,and si NEAT1+miR-195-5p inhibitor(miR-195-5p inhibitor)group,while untreated HBMSCs were used as the controls and transfected for osteogenic induction.The qRT-PCR,CCK-8 method(OD450),Alizarin Red staining,alkaline phosphatase(ALP)assay kit,and Western Blot,respectively,were used to detect the expression levels of NEAT1,miR-195-5p,TXNIP mRNA,cell viability,mineral-ization ability,ALP activity,the expression levels of TXNIP,BMP2,and OCN proteins of HBM-SCs.The dual luciferase assay was applied to verify the targeting relationship of miR-195-5p with NEAT1 and TXNIP.Results:The expression levels of NEAT1 and TXNIP mRNA significantly de-creased after osteogenic differentiation,while the expression of miR-195-5p significantly increased(P<0.05).Compared with those in the si NC group and control group,obvious nodule formation was found in the si NEAT1 group,the expression of NEAT1 and TXNIP were significantly reduced,the expression of miR-195-5p,OD450 values at 24 and 48 hours,ALP activity,the expression of BMP2,and OCN proteins were significantly increased(P<0.05).Compared with those in the mimics NC group,obvious nodule formation was found in the miR-195-5p mimics group,the expression of TXNIP was significantly reduced,the expression of miR-195-5p,OD450 values after 24 and 48 hours,ALP activity,the expression of BMP2,and OCN proteins were significantly increased(P<0.05),but no significant changes of NEAT1 expression was found(P>0.05).Compared with those in the si NEAT1+inhibitor NC group,the si NEAT1+miR-195-5p inhibitor group had few nodules and light-er color,the expression of TXNIP protein and mRNA was significantly increased,the expression of miR-195-5p,OD450 values after 24 and 48 hours,ALP activity,the expression of BMP2,and OCN proteins were significantly reduced(P<0.05).However,the changes in NEAT1 expression were not statistically significantl too(P>0.05).The miR-195-5p had targeted relationships with NEAT1 and TXNIP,respectively.Conclusion:LncRNA NEAT1 promotes osteogenic differentiation of HBM-SCs by up-regulating the miR-195-5p/TXNIP axis.
LncRNA NEAT1miR-195-5p/TXNIP AxisHuman Bone Marrow Mesenchy-mal Stem CellsOsteogenic Differentiation
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维普
