【Animal Modeling】-Protective effect of estrogen on oxidative stress damage of mouse osteoblast MC3T3-E1 and its mechanism

  Objective: To investigate the protective effect and mechanism of estrogen on oxidative stress injury of mouse osteoblast MC3T3-E1.

  Method: MC3T3-E1 cells cultured in vitro were used as a blank control group, H2O. Corresponding group (300 μmol/L), H2O2+estrogen 0.1 μmol/L group, H, O2+estrogen 1 μmol/L group, H, O, + estrogen 10 μmol/L group and H, O, +AC 1 mmol /L group. Incubation with drug concentration. CCK-8 was used to detect cell proliferation activity in each group, and flow cytometry was used to detect cell apoptosis rate and mitochondrial membrane potential. The kit can detect malondialdehyde (MDA) and superoxide dismutase (SOD) levels. Fluorescence probe method detects cells. Reactive oxygen species (ROS) levels; detect protein expression in WesternBlot Smad5, Runx2, Bax, Bcl-2 and cleavedCaspase-3. and

  Result: Compared with the blank control group, the proliferation activity of osteoblasts in the H, O, and groups of mice was significantly reduced (Pu003c0.05), and the apoptosis rate and the rate of JC-1 positive cells were significantly increased. (Pu003c MDA and ROS levels are significantly increased (Pu003c0.05), SOD activity is significantly reduced (Pu003c0.05), and the expression of Smad5, Runx2 and Bcl-2 proteins are significantly reduced (Pu003c0.05) u003c0.05), And Bax and cleavedCaspase-3 protein expression increased significantly (Pu003c0.05); H, 0.H, compared with the group. O0, + estrogen 1μmol/L group, H, O, + estrogen 10 cell proliferation activity μmol/L group and H, O, +NAC 1mmol/L group significantly increased (Pu003c0.05), cell apoptosis and JC The proportion of -1 positive cells was significantly reduced (Pu003c0.05), the levels of MDA and ROS were significantly reduced (Pu003c0.05), SOD activity was significantly increased (Pu003c0.05), and the expression of Smad5, Runx2 and Bcl-2 proteins were significantly increased (Pu003c0.05), while Bax and cleavedCaspase-3 protein expression was significantly reduced (Pu003c0.05). Pu003c0.05).

  Conclusion: Estrogen may reduce the level of ROS in MC3T3-E1 cells damaged by oxidative stress and improve their differentiation ability by activating the expression of Smad5/Runx2 signal axis.