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目的探讨不同浓度绞股蓝皂苷(GPs)对体外培养的神经前体细胞(NPCs)增殖能力的影响。方法从孕14 d大鼠胚胎端脑分离NPCs,体外贴壁培养7 d传代。传代第1代细胞培养3 d,免疫荧光法鉴定NPCs纯度后进行分组实验。加不同浓度GPs(0、25、50、100、200、400μg/mL)作用48h后再次鉴定NPCs纯度,采用MTT法检测细胞活力、细胞计数绘制生长曲线、5-溴脱氧尿嘧啶核苷(BrdU)掺入法检测细胞增殖能力、Western blot法检测细胞内增殖细胞核抗原(PCNA)表达情况。结果传代第1代培养3 d的NPCs纯度达97%,GPs作用后不影响NPCs纯度,可使细胞活性增强,生长速度加快,BrdU阳性率增高,PCNA表达水平上调。结论GPs可通过提高PCNA的表达量,促进体外培养的NPCs增殖,100μg/mL为GPs最佳作用浓度。
Objective To investigate the effects of different concentrations of gypenosides (GPs) on the proliferation of neural progenitor cells (NPCs) cultured in vitro. Methods NPCs were isolated from embryonic telencephalon of rats on the 14th day of pregnancy and cultured in vitro for 7 days. Passage 1 cells were cultured for 3 days. Grouped experiments were performed after the purity of NPCs was determined by immunofluorescence. The purity of NPCs was determined again after 48 h of addition of different concentrations of GPs (0, 25, 50, 100, 200, 400 μg/mL). Cell viability was measured by MTT assay, growth curves of cell counts, and 5-bromodeoxyuridine (BrdU) were determined. The cell proliferation ability was detected by the incorporation method, and the expression of proliferating cell nuclear antigen (PCNA) in the cells was detected by Western blot. Results The purity of NPCs reached 97% after 3 days of culture in passage 1, GPs did not affect the purity of NPCs, and the activity of cells increased, the growth rate increased, the BrdU positive rate increased, and the expression of PCNA increased. Conclusion GPs can increase the expression of PCNA and promote the proliferation of NPCs cultured in vitro. 100 μg/mL is the best concentration of GPs.