目的:观察辛伐他汀对骨髓源性平滑肌祖细胞(SPC )分化及增殖的影响,筛选新一代包被洗脱支架药物。方法:采用密度梯度离心法从大鼠骨髓获取单个核细胞,接种于纤维连接素包被的培养板,培养8d后,以平滑肌肌动蛋白(α-SM A )免疫荧光染色鉴定骨髓源性SPC ,并在倒置荧光显微镜下计数。收集培养8d单个核细胞、贴壁细胞,分别加入不同浓度辛伐他汀(0.01-10μmol/L )培养8d、24h。分别采用氚标记胸苷(3 H-TdR)摄入法、改良的Boyden小室和黏附能力测定实验观察SPC的分化、增殖、迁移和黏附能力。结果:与对照组(无辛伐他汀干预)相比,0.01μmol/L辛伐他汀显著抑制骨髓单个核细胞向SPC分化[(85±4)个比(79±5)个]、增殖[(4070±184)个比(3833±126)个]、迁移[(44±3)个比(39±3)个]和黏附[(59±5)个比(52±4)个], P均<0.05,且随辛伐他汀浓度增加S PC数量显著减少( P<0.01)。结论:辛伐他汀可抑制骨髓源性平滑肌祖细胞分化、增殖、迁移及黏附能力。
Objective:To observe influence of simvastatin on differentiation ,proliferation ,migration and adhesion of marrow-derived smooth muscle progenitor cells (SPCs) and screen coated eluting stent drugs of new generation . Methods :The mononuclear cells (MNCs) were isolated from rat marrow by density gradient centrifugation method , and then plated on fibronectin-coated culture dishes ,after culture 8d ,marrow-derived SPCs were identified by α-smooth muscle actin (α-SMA) immunofluorescent staining and counted under inverted fluorescence microscope .The MNCs and adhesion cells were treated with simvastatin (0.01-10 μmol/L) respectively for 8 d and 24h .SPCs pro-liferation ,migration and adhesion were observed by Tritium thymidine (3 H-TdR) intake method ,modified Boyden chamber assay and adhesion assay .Results:Compared with control group (no simvastatin intervention ) ,0.01μmol/L simvastatin significantly inhibited the MNCs differentiation towards SPCs [ (85 ± 4) vs .(79 ± 5)] ,proliferation [ (4070 ± 184) vs .(3833 ± 126)] ,migration [ (44 ± 3) vs .(39 ± 3)] and adhesion of SPCs [ (59 ± 5) vs .(52 ± 4)] , P〈0.05 all ,and number of SPCs significantly reduced along with simvastatin concentration increased (P〈0.01) . Conclusion:Simvastatin could inhibit the differentiation ,proliferation ,migration and adhesion of marrow-derived smooth muscle progenitor cells .