Rosuvastatin regulates osteogenic differentiation of mesenchymal stem cells through phosphatidylinositol-3-kinase/protein kinase B

Abstract

Objective To investigate the effects of rosuvastatin on proliferation, osteogenic differentiation and phosphatidylinositol-3-kinase (PI3K)/protein kinase B (Akt) expression of mesenchymal stem cells (MSCs). Methods MSCs were isolated. Rosuvastatin at concentrations of 1×10-11, 1×10-9 and 1×10-7 mol/L were added to the experimental groups, and dimethylsurfoxide (DMSO) served as control. Cell proliferation was measured by methyl thiazol tetrazolium (MTT) assay. After 3, 7, 14 and 21 days of osteogenic induction, alkaline phosphatase (ALP) quantitative analysis, alizarin red staining and quantitative analysis, real-time quantitative polymerase chain reaction (Real-time PCR) and Western blotting were performed. T-test was used for comparison between two groups, and ANOVA for more than two groups. Results At 24 h and 48 h, the proliferation of MSCs in high concentration group was significantly higher than in the control group (24 h: 3.57±0.24 vs. 3.14±0.14, t=-3.851, P<0.05; 48 h: 4.19±0.18 vs. 3.44±0.11, t=-6.780, P<0.05). Until 72 h, the proliferation with 1×10-9 mol/L rosuvastatin was also significantly increased (5.42±0.13 vs. 4.47±0.16, t=-8.700, P<0.05). At 3rd day after induction culture, 1×10-7 mol/L rosuvastatin significantly enhanced the genes expression of BMP2, Runx 2, OPN 2, OPN and ColⅠ (BMP2: 2.1±0.2 vs. 1.0±0.2, t=-6.736, P<0.05; Runx2: 5.2±0.6 vs. 1.0±0.1, t=-11.959, P<0.05; OPN: 1.8±0.4 vs. 1.0±0.2, t=-3.098, P<0.05; ColⅠ: 1.9±0.3 vs. 1.0±0.3, t=-3.674, P<0.05); 1×10-7 mol/L rosuvastatin could increase the expression of p-Akt/PI3K. The ALP activity after adding 1×10-9 or 1×10-7 mol/L rosuvastatin was significantly increased at 14th and 21st day (14 d: 12.07±1.67, 18.32±2.26 vs. 3.43±0.36, F=7.200, P<0.05; 21 d: 10.74±1.27, 14.71±3.18 vs. 5.76±0.63, F=6.489, P<0.05). The quantitative results of alizarin red were similar to those of the staining. Meanwhile, the quantitative values of alizarin red in the 1×10-9 and 1×10-7 mol/L groups were also significantly increased (14 d: 5.6±0.8, 6.2±1.2 vs. 1.0±0.2, F=5.600, P<0.05; 21 d: 5.8±1.1, 7.1±1.8 vs. 2.4±0.3, F=5.956, P<0.05). Conclusion Rosuvastatin promotes proliferation and osteogenic differentiation of MSCs, which may be related to PI3K/Akt signaling. Key words: Rosuvastatin; Phosphatidylinositol-3-kinase/protein kinase B signaling pathway; Mesenchymal stem cells; Osteogenic differentiation