Berberine Regulates the Proliferation and Osteogenic Differentiation of HumanPeriodontal Ligament Stem Cells through PI3K/AKT Signaling Pathway

This study aims to investigate the effect and potential mechanism of berberine on the proliferation and osteogenic differentiation of hPDLSCs (human periodontal ligament stem cells). hPDLSCs werecultured in vitro, and were divided into blank Con (control) group, LY (PI3K/AKT signaling pathway inhibitor LY294002) group, Ber (berberine) group and Ber+LY (berberine+PI3K/AKT signaling pathway inhibitorLY294002) group. CCK-8 (cell counting kit-8) method was used to detect the proliferation activity of hPDLSCs.Flow cytometry was used to detect cell cycle distribution. Enzyme-linked immunosorbent assay was used to detect ALP (alkaline phosphatase) activity. qRT-PCR (real-time fluorescent quantitative PCR) was used to analysisthe expression of PCNA (proliferating cell nuclear antigen), Cyclin D1, OCN (osteocalcin), POSTN (periostin)and OPN (osteopontin). Western blot was used to detect the expression of PCNA, Cyclin D1, OCN, POSTN, OPN and PI3K/AKT signaling pathway-related proteins. Compared with the Con group, the proliferation activity of hPDLSCs in the Ber group, ALP activity, S phase and G2/M phase cell ratio, PCNA, Cyclin D1, OCN,POSTN, OPN, p-PI3K and p-AKT expression levels were significant increased, the proportion of cells in G0/G1phase was significantly reduced (P<0.05), while the above indicators in the LY group showed opposite changes(P<0.05). Compared with the Ber group, the proliferation activity of hPDLSCs in the Ber+LY group, the activityof ALP, the ratio of cells in S phase and G2/M phase, the expression levels of PCNA, Cyclin D1, OCN, POSTN,OPN, p-PI3K and p-AKT were significantly reduced, and the proportion of cells in G0/G1 phase was significantlyincreased (P<0.05). Berberine can promote the proliferation of hPDLSCs and induce the osteogenic differentiation of hPDLSCs by activating the PI3K/AKT signaling pathway.