目的研究淫羊藿苷对过载损伤后成骨细胞的凋亡和细胞骨架的影响。方法应用4点弯曲加载装置模拟过载损伤力学环境,建立过载损伤模型,依据力学加载前后是否加入药物干预,实验分为空白对照组、单纯给药组、单纯损伤组、预防给药组和损伤治疗组。应用细胞流式术进行细胞凋亡检测。应用特异性荧光染料分别标记微丝蛋白和细胞核,在激光共聚焦显微镜下观察细胞骨架的变化。结果与对照组比,过载损伤组的凋亡率最高,单纯给药组最低(P〈0.05);在过载处理组中,与单纯过载损伤组比,预防给药组的凋亡率最低(P〈0.05)。单纯过载损伤组的细胞皱缩变形,微丝排列紊乱,骨架排列疏松,轮廓模糊,骨架甚至出现破碎等现象。预防给药组细胞骨架形态变化不大,细胞核未见明显变化。结论淫羊藿苷可以在一定程度上抑制过载损伤后成骨细胞的凋亡,维持细胞骨架稳定。
Objective To study the effect of 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 the control group,the apoptosis rate of the damage group was the highest,and the icariin group was the lowest(P〈0.05).Compared with the 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.