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目的研究老鹳草素对小鼠骨髓基质干细胞(BMSCs)增殖及成骨分化的影响。方法分离、培养小鼠BMSCs,细胞分4组:空白组(加完全培养基),3个剂量老鹳草素实验组(老鹳草素,1×10~(-7),1×10~(-8),1×10~(-9)mol·L~(-1)),CCK-8法检测细胞增殖,流式细胞仪检测细胞凋亡和细胞周期变化,碱性磷酸酶(ALP)试剂盒检测细胞ALP活性,茜素红染色检测成骨分化过程中钙结节形成数量。结果在成骨诱导培养1,3,5,7 d,老鹳草素各剂量组明显促进BMSCs的增殖,同时细胞凋亡率显著降低,呈剂量依赖性。第3天,3个剂量老鹳草素实验组A_(450)值分别为0.41±0.06,0.37±0.04,0.33±0.05,较空白组(0.29±0.03)显著增加(P<0.05),细胞凋亡率分别为(2.53±0.40)%,(3.02±0.46)%,(3.71±0.54)%,较空白组(4.22±0.58)%显著降低(P<0.05);与空白组比较,G_0/G_1期细胞比例显著降低,S期细胞比例及细胞ALP活性显著增加(均P<0.05)。在成骨诱导培养21 d,与空白组比较,各剂量老鹳草素组的矿化结节数量显著增加(P<0.05)。结论老鹳草素通过调控BMSCs细胞周期、抑制凋亡促进BMSCs的增殖,还明显促进BMSCs向成骨方向分化。
Objective To study the effects of geranium on the proliferation and osteogenic differentiation of mouse bone marrow stromal stem cells (BMSCs). Methods BMSCs were isolated and cultured. The cells were divided into 4 groups: blank group (complete medium), three doses of geraniol experimental group (geraniol, 1 × 10 -7, 1 × 10 ~ (-8), 1 × 10 -9 mol·L -1). Cell proliferation was detected by CCK-8 assay. Apoptosis and cell cycle were detected by flow cytometry. ALP ) Kit was used to detect the cell ALP activity. Alizarin red staining was used to detect the number of calcium-forming nodules during osteogenic differentiation. Results After osteogenic induction of culture for 1, 3, 5 and 7 d, the dose of geraniol significantly promoted the proliferation of BMSCs, and the apoptosis rate was significantly decreased in a dose-dependent manner. On the third day, the values of A 450 in the three doses of gentianin experimental group were 0.41 ± 0.06, 0.37 ± 0.04 and 0.33 ± 0.05, respectively, which were significantly higher than those in the blank group (0.29 ± 0.03) (P <0.05) (2.53 ± 0.40)%, (3.02 ± 0.46)% and (3.71 ± 0.54)%, respectively, which were significantly lower than those in the blank group (4.22 ± 0.58)% (P <0.05) The proportion of senescent cells was significantly decreased, the proportion of cells in S phase and the activity of ALP increased significantly (all P <0.05). Compared with the blank group, the number of mineralized nodules in each dose of geranium significantly increased (P <0.05) at 21 days after osteogenic induction. Conclusion Geranium could promote the proliferation of BMSCs by regulating the cell cycle and inhibiting the apoptosis of BMSCs, and also promote the differentiation of BMSCs into osteoblasts.