|
Objective To investigate the different concentrations of pioglitazone on proliferation and apoptosis of osteoblasts, further understanding of the mechanism of apoptosis of osteoblasts and provide some experimental basis for the clinical use of drugs. Methods (1) MC3T3-E1 mouse embryos into bone cell lines as experimental subjects, divided into six groups, groups of four parallel, respectively, to give the 5,10,20,30,40 μM pioglitazone ketone intervention control group dose of dimethyl sulfoxide, 24 hours after the MTT assay osteoblast activity (2) will be divided into 6 groups, each group of three parallel, respectively, given the 5,10,20,30,40 μM pioglitazone intervention The control group intervention after 24 hours by flow cytometry proportion of each group the rate of apoptosis and cell cycle phase (3) be divided into five groups, each group of six parallel, the same dose of dimethyl sulfoxide were given the 10,20,30,40 μM pioglitazone intervention, control group 24 hours after the same dose of dimethyl sulfoxide, osteoblasts Bcl-2 and Bax protein expression detected by immunohistochemistry method. (1) With the concentration of pioglitazone increased osteoblast activity gradually decreases in the concentration range of 0-30μM were statistically different between the groups (p lt; 0.05), in a dose-dependent decrease, 30,40 μM The two groups showed no significant difference (p gt; 0.05). (2) a) cell cycle phase ratio: compared with the control group, each the concentration group Go/G1 cells are increased, which the 5μM group and the control group was not statistically different (p gt; 0.05); G2M phase cells compared with control group increased 20, 30, 40 μM and control groups had no statistical difference (p gt; 0.05); decrease in S phase cells concentration ≤ 30μM time, compared with the control group was significantly decreased (p lt; 0.01) less than 40μM the control group (p lt; 0.05). b) osteoblast apoptosis rate lower than the control group (p lt; 0.05): 5,10 μM, 20μM slightly lower than the control group, but not statistically significant, 30, 40 μM compared with the control group was significantly higher (p lt; 0.01 ). (3) Bax expression and control group when compared 10μM significantly reduced (p lt; 0.01), 20μM close to the control group (p gt; 0.05), 30, 40 μM compared with the control group significantly increased (p lt; 0.01); of Bc1 -2 expression was significantly increased (p lt; 0.01) concentration ≤ 20μM 30μM stronger than the control group (p lt; 0.05) the 40μM expression significantly reduced compared with the control group (p lt; 0.01). (4) For the strength and rate of apoptosis Bax/Bc1-2 relative expression correlation analysis, both showed a significant positive correlation (n = 15, r = 0.796, P lt; 0.001). Conclusion Pioglitazone promote osteoblast apoptosis at low concentrations (lt; 20μM) inhibition of osteoblast apoptosis protective effect on cells, higher concentrations (30μM, 40μM), Bax/Bcl-2 involved in the mechanism of apoptosis and may play a key role. Overall can inhibit the DNA synthesis of osteoblasts, inhibits cell proliferation, resulting in cell activity decreased.
|