普伐他汀对骨髓间充质干细胞迁移和黏附能力的影响

目的观察普伐他汀干预对人骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMMSCs)迁移和黏附能力的影响及其相关细胞信号传导通路。方法使用普伐他汀干预体外培养的第6代人BMMSCs,Western blotting检测作用前后ERK1/2、p38MAPK及PI3K/Akt通路蛋白的表达情况。将10μmol/L普伐他汀预处理人BMMSCs1h后,通过Transwell小室进行细胞迁移实验,并进行细胞黏附性测定,进一步用特异的细胞信号通路抑制剂或激动剂阻断或激活ERK1/2、p38MAPK及PI3K/AKT途径,观察其对普伐他汀作用的影响。结果普伐他汀可使人BMMSCs的PI3K/Akt通路磷酸化水平升高,抑制p38MAPK通路磷酸化水平,而其对人BMMSCs的ERK通路和总Akt、总p38MAPK水平无显著影响。经普伐他汀作用后迁移细胞显著增多(P<0.05),Ly294002预处理后这种作用消失,anisomycin预处理对这种作用影响不明显。普伐他汀作用后贴壁细胞显著增多(P<0.05),但Ly294002或anisomycin预处理对这种作用影响均不明显。结论普伐他汀具有增强人BMMSCs迁移和黏附能力的作用。其增强迁移能力的作用与激活人BMMSCs的PI3K/Akt通路有关。但是它对黏附能力的作用与PI3K/Akt和p38MAPK通路均无关。

Effects of pravastatin on migration and adhesion of human bone marrow mesenchymal stem cells

Objective To investigate the effects and related cell signaling pathways of pravastatin on migration and adhesion of human bone marrow mesenchymal stem cells （BMMSCs）. Methods After being cultured in vitro, passage 6 human BMMSCs were treated with pravastatin. Phosphorylation of ERK 1/2, p38MAPK and PI3K/Akt were detected by western blotting. After 24 hours of pretreatment with 10 μmol/L pravastatin, cell migration assay was performed in Transwell chambers, and adhesion assay was performed by plastic dishes. Results Pravastatin could increase phosphorylation of PI3K/Akt pathway of human BMMSCs, but reduce phosphorylation of p38MAPK. It had no obvious effect on ERK1/2 pathway and total proteins of Akt and p38MAPK. The number of cells crossing the filter was significantly increased in the pravastatin group （P 0.01）. This effect was completely inhibited by concomitant incubation with Ly294002. After treated with pravastatin, adherent cells increased significantly （P 0.01）. Neither Ly294002 nor anisomycin could inhibit this effect. Conclusions Pravastatin can activate PI3K/Akt pathway and inhibit p38 MAPK pathway of human BMMSCs. Activation of PI3K/Akt pathway was involved in the effect of pravastatin on cell migration. The effect of pravastatin on adhesion capacity had nothing to do with PI3K/Akt and p38MAPK pathway.