Ginsenoside Rg1 Induces the Proliferation and Differentiation of Bone Marrow Mesenchymal Stem Cells into Nucleus Pulposusk-like Cells Through the miR-138-5p/SIRT1 Axis

OBJECTIVE To explore the mechanism of ginsenoside Rg1 induced differentiation of rat bone marrow mesenchymal stem cells(BMSCs) into nucleus pulposus like cells(NPCs).METHODS Isolation and culture of BMSCs from adult SD rats. The effects of different concentrations of ginsenoside Rg1(0, 2.5, 5, 10, 20 μmol·L^-1) on the viability of BMSCs were detected by CCK-8 assay. The cells were divided into blank group, 5 μmol·L^-1 ginsenoside Rg1 intervention group and 10 μmol·L^-1 ginsenoside Rg1 intervention group. After 48 h of culture, and then the apoptosis rate was determined by flow cytometry, the expression of NPCs marker proteins collagen type 2(COL2), Aggrecan and paired box gene 1(Pax1) was detected by Western blotting. BMSCs were divided into blank group, ginsenoside Rg1 group, ginsenoside Rg1+NC mimic group, and ginsenoside Rg1+miR-138-5p mimic groups, followed by the detection of the effect of miR-138-5p on the BMSC differentiation induced by ginsenoside Rg1. The target relationship between miR-138-5p and sirtuin-1(SIRT1) was detected by dual luciferase reporter and RNA binding protein immunoprecipitation assays. pcDNA-SIRT1 was transfected into BMSCs to detect its effect on the differentiation of BMSCs induced by miR-138-5p.RESULTS The 2.5, 5 and 10 μmol·L^-1 ginsenoside Rg1 enhanced BMSCs viability in a dose-dependent manner(P < 0.05). Compared with the blank group, 5 μmol·L^-1 and 10 μmol·L^-1 ginsenoside Rg1 intervention groups inhibited cell apoptosis(P < 0.05) and promoted the expression of NPCs marker proteins in a dose-dependent manner(P < 0.05). Compared with the ginsenoside Rg1 intervention group, the apoptosis rate of BMSCs in the ginsenoside Rg1+miR-138-5p group was increased(P < 0.01) and the expression of NPC marker proteins was decreased(P < 0.01). Moreover, SIRT1 was a target gene of miR-138-5p. Compared with the miR-138-5p mimic+pcDNA-3.1 group, the apoptosis rate of BMSCs in the miR-138-5p mimic+pcDNA-SIRT1 group was decreased(P < 0.01), and the expression of NPCs marker proteins was increased(P < 0.01).CONCLUSION Ginsenoside Rg1 can induce BMSCs to proliferate and differentiate into NPCs by regulating the miR-138-5p/SIRT1 axis.