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目的应用流式细胞仪检测细胞衰老的特征。方法选取3月龄健康雄性大鼠的原代系膜细胞,通过传代形成细胞衰老模型。光镜下观察细胞各代的形态结构,并用衰老相关β-半乳糖苷酶活性(SA-β-gal)染色明确细胞衰老程度;采用流式细胞仪分析细胞衰老的特征,包括自发荧光、细胞大小(前向角散射光,FSC)及细胞周期。结果随传代次数的增加,大鼠肾小球系膜细胞形态变扁平、肥大,且SA-gal活性增强(P<0.05),流式细胞仪结果也显示,细胞自发荧光有显著增加(P<0.05);FSC增大(P<0.05);细胞周期趋向停滞于G0/G1期,细胞增殖能力下降。结论系膜细胞在复制性衰老过程中,伴随着细胞肥大、细胞周期异常,增殖能力下降及细胞内自发荧光产物增加,流式细胞仪能很好地检测到这些改变,因此它是一种检测细胞衰老指标的良好方法。
Objective To detect the characteristics of cellular senescence by flow cytometry. Methods Primary mesangial cells of 3-month-old healthy male rats were selected and passaged to form a cell aging model. The morphology of the cells was observed under light microscope, and the cell senescence was determined by aging-related β-galactosidase activity (SA-β-gal) staining. Cell senescence characteristics were analyzed by flow cytometry, including autofluorescence, Size (Forward Scattered Light, FSC) and Cell Cycle. Results The mesangial cells became flattened and hypertrophy with SA-gal activity increased (P <0.05). The results of flow cytometry also showed that the autofluorescence of cells increased significantly (P < 0.05); FSC increased (P <0.05); cell cycle tended to arrest in G0 / G1 phase and cell proliferation decreased. Conclusions Mesangial cells are associated with cell hypertrophy, cell cycle abnormalities, decreased proliferative ability and increased intracellular autofluorescence during replication-induced senescence. Flow cytometry can detect these changes well, so it is a test A good way to measure cellular senescence.