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0.1,1,10μg·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RhoA和Cofilin的mRNA表达量。结果与空白对照组比较,100μg·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0.1μg·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),0.1~10μg·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RhoA和Cofilin的mRNA表达有关。
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~(-1)0 μg·mL~(-1) icariin(P<0.05 or P<0.01), while0.1 μg·mL~(-1) icariin had no significant effect. Compared with the model group, 1~(-1)0 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~(-1)0 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.
引文
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