淫羊藿苷对脂多糖诱导成骨细胞骨架F-actin损伤的保护作用

Protective effect of Icariin on Lipopolysaccharide-induced cytoskeleton F-actin injury

关键词:  淫羊藿苷  脂多糖  细胞骨架  F-肌动蛋白

Key words:  icariin  lipopolysaccharide  cytoskeleton  F-actin

摘要:

目的 考察淫羊藿苷对成骨细胞骨架F-actin损伤的保护作用及其对相关信号通路的调控。方法 采用脂多糖诱导大鼠成骨细胞损伤模型。实验分为空白对照组、模型组和0.1，1，10 μg·mL^-1淫羊藿苷组，MTT法观察淫羊藿苷对成骨细胞活性的影响，TRITC-Phalloidin荧光染色观察淫羊藿苷对细胞骨架的影响，ELISA法测定各组成骨细胞中F-actin的含量，RT-PCR法测定细胞骨架Rho A和Cofilin的mRNA表达量。结果 与空白对照组比较，100 μg·mL^-1脂多糖能显著降低成骨细胞存活率（P<0.01）；与模型组比较，1~10μg·mL^-1淫羊藿苷预处理后能显著提高细胞存活率（P<0.05或P<0.01）；而0.1 μg·mL^-1淫羊藿苷则无显著影响。与模型组比较，1~10μg·mL^-1淫羊藿苷能显著抑制成骨细胞F-actin解聚（P<0.05），抑制Rho A的mRNA表达（P<0.05或P<0.01），0.1~10 μg·mL^-1淫羊藿苷能抑制Cofilin的mRNA表达（P<0.01）。结论 淫羊藿苷对脂多糖诱导的细胞骨架F-actin损伤具有保护作用，其机制与抑制细胞骨架相关因子Rho A和Cofilin的mRNA表达有关。

Abstract:

OBJECTIVE To investigate the protective effect of icariin on F-actin damage of osteoblast cytoskeleton and the regulation of related signaling pathways. METHODS Lipopolysaccharide(LPS) was used to induce osteoblasts to establish an injury model in vitro. Experimental group:control group, model group and 0.1, 1, 10 μg·mL^-1 icariin groups. MTT method was used to observe the effect of icariin on the activity of osteocytes. TRITC-Phalloidin fluorescence staining was used to observe the effect of icariin on cytoskeleton. ELISA method was used to determine the content of F-actin in osteocytes. RT-PCR method was used to determine the expression of cytoskeleton-related factors Rho A and Cofilin mRNA. RESULTS Compared with the control group, 100 μg·mL^-1 LPS could significantly reduce the survival rate of osteoblasts(P<0.01). Compared with the model group, the survival rate could significantly improve after pretreatment with 1-10 μg·mL^-1 icariin(P<0.05 or P<0.01), while 0.1 μg·mL^-1 icariin had no significant effect. Compared with the model group, 1-10 μg·mL^-1 icariin could significantly inhibit F-actin depolymerization in osteoblasts(P<0.05), inhibit the mRNA expression of Rho A (P<0.05 or 0.01), and icariin at concentration of 0.1-10 μg·mL^-1 could inhibit the mRNA expression of Cofilin (P<0.01).CONCLUSION Icariin has protective effect on LPS-induced cytoskeleton F-actin injury model, and its mechanism may be related to inhibiting the expression of cytoskeleton-related factors Rho A and Cofilin mRNA.