Objective To study the effect of the icariin on apoptosis and cytoskeleton of osteoblasts in response to overload damage. Methods The four-point bending loading device was used to simulate the mechanical environment of overload damage and establish the cell overload damage model. According to whether the drugs were added before or after mechanical loading, the experiment was divided into blank control group, icariin group, damage group, damage prevention group and damage treatment group. Cell apoptosis was detected by flow cytometry. The specific fluorescent dyes were used to label the actin filament and the nucleus, and the changes of cytoskeleton were observed under laser scanning confocal microscope. Results Compared with control group, the apoptosis rate of damage group was the highest, and the icariin group was the lowest (P<0.05). Compared with damage group, the apoptosis rate of the damage prevention group was the lowest (P<0.05). The damage group showed cell shrinkage deformation, microfilaments disorganization, loosely arranged skeleton with vague outline, even broken skeleton. The morphological changes of cytoskeleton in damage prevention group were not significant, and there was no obvious change in cell nucleus. Conclusions Icariin can inhibit the apoptosis of osteoblasts after overload injury and maintain the stability of cytoskeleton to some extent.