论文部分内容阅读
目的观察雷帕霉素对骨髓源性平滑肌祖细胞增殖及分化的影响,探讨影响动脉粥样硬化的骨髓源性平滑肌祖细胞与雷帕霉素洗脱支架抑制新内膜增生的关系。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,加入不同浓度雷帕霉素(0.01~100μg/L),培养12天后,-αSMA免疫荧光染色鉴定骨髓源性平滑肌祖细胞,并在倒置荧光显微镜下计数。收集培养8天贴壁细胞,分别加入不同浓度雷帕霉素(0.1~200μg/L)培养24 h。分别采用MTT比色法、改良的Boyden小室和粘附能力测定实验观察平滑肌祖细胞的增殖、迁移和粘附能力。结果雷帕霉素显著抑制骨髓单个核细胞分化为平滑肌祖细胞,0.1μg/L雷帕霉素作用12天,平滑肌祖细胞减少了70.9%±4.5%(P<0.01)。雷帕霉素也显著抑制平滑肌祖细胞增殖,其抑制作用随雷帕霉素浓度增加而增加,1μg/L雷帕霉素作用24 h使平滑肌祖细胞数量减少27.1%±4.5%(P<0.01)。0.1~200μg/L雷帕霉素显著抑制平滑肌祖细胞的粘附和迁移能力。结论雷帕霉素可抑制平滑肌祖细胞分化,并抑制平滑肌祖细胞增殖及迁移能力。
Objective To observe the effect of rapamycin on the proliferation and differentiation of bone marrow derived smooth muscle progenitor cells and to explore the relationship between the effect of rapamycin eluting stent and neointimal hyperplasia on atherosclerotic myeloid derived smooth muscle progenitor cells. Methods Mononuclear cells were obtained from bone marrow of rats by density gradient centrifugation. The cells were seeded onto fibronectin-coated plates and incubated with different concentrations of rapamycin (0.01-100 μg / L) for 12 days. After α-SMA immunofluorescence staining Bone marrow derived smooth muscle progenitor cells were identified and counted under an inverted fluorescence microscope. Adherent cells were harvested for 8 days and cultured in different concentrations of rapamycin (0.1-200 μg / L) for 24 h. MTT assay, modified Boyden chamber and adhesion assay were used to observe the proliferation, migration and adhesion of smooth muscle progenitor cells. Results Rapamycin significantly inhibited the differentiation of bone marrow mononuclear cells into smooth muscle progenitor cells. After treated with 0.1μg / L rapamycin for 12 days, smooth muscle progenitor cells decreased by 70.9% ± 4.5% (P <0.01). Rapamycin also significantly inhibited the proliferation of smooth muscle progenitor cells, the inhibitory effect increased with the increase of rapamycin concentration, 1μg / L rapamycin for 24 h reduced the number of smooth muscle progenitor cells by 27.1% ± 4.5% (P <0.01 ). 0.1 ~ 200μg / L rapamycin significantly inhibited the adhesion and migration of smooth muscle progenitor cells. Conclusion Rapamycin can inhibit the differentiation of smooth muscle progenitor cells and inhibit the proliferation and migration of smooth muscle progenitor cells.