Effect of LIMK2 Inhibition on the Reconstruction of Cytoskeleton

Objective: To investigate the role of LIMK2 in the cytoskeleton reconstruction of osteoblast induced by fluid shear stress. Methods: Mouse primary osteoblasts were divided into 4 groups and named group 1,2,3 and 4. Cells in group 1 and group 3 were transfected with negative control RNA which has no specific silence effect; cells in group 2 and group 4 were transfected with siRNA which can silence the expression of LIMK2. After transfection, cells were digested and cultured in the cell loading device. Cells in group 1 and group 2 were loaded by fluid shear stress while group 3 and group 4 were not. Afterward, the cytoskeleton was observed with LSCM (laser scanning confocal microscope). Furthermore, the photos of the cells were taken and the mean fluorescence density of each cell in the photos was measured and recorded. Results: Compared with group 1, the fluorescence of cytoskeleton in group 2, group 3, and group 4 was decreased to 28.9%, 45.7%, and 33.0%, respectively. Diversity between group 1 and the rest three groups is significant at the .05 level; diversity between group 3 and the rest three groups is significant at the .05 level and diversity between group 2 and group 4 is not significant at the .05 level. Conclusion: The reconstruction of cell cytoskeleton will be decreased when the LIMK2 expression of mouse primary osteoblasts were inhibited. It indicates that LIMK2 plays important role in the reconstruction of cytoskeleton induced by fluid shear stress, and without LIMK2, the pathway will be blocked.