人肝细胞生长因子对体外培养的黑素细胞生物学活性的影响

Effects of recombinant human hepatocyte growth factor on cultured human melanocytes in vitro

目的 探讨人肝细胞生长因子（rhHGF）对体外培养正常人黑素细胞生物学活性的影响。方法分别用不同浓度的rhHGF（0．1，1，10，20，60，80，100μg／L）作用于体外培养的正常人黑素细胞，采用噻唑蓝（MTT）法检测细胞增殖及酪氨酸酶活性，用倒置显微镜对比观察不同时期rhHGF（60μg／L）处理组黑素细胞形态的改变，并用流式细胞仪分析黑素细胞周期及凋亡情况。结果0．1～100μg／L rhHGF有促进黑素细胞增殖作用，在20，60，80μg／L浓度时促增殖比较显著（P〈O．01），rhHGF各浓度组均增加黑素细胞酪氨酸酶活性（P〈0．01或P〈0．05）；浓度为60μg／L的rhHGF能使黑素细胞数目增多、胞体增大、树突增多，并有促进黑素细胞由GdG。期进入S期及抑制细胞凋亡的作用。结论 rhHGF可增强体外培养的人黑素细胞的生物学活性。

Objective To investigate the effects of recombinant human hepatocyte growth factor （rhHGF） on biologic activity of human melanocytes/n vitro. Methods Human melanocytes were primarily cultured. After certain passages, the cells were treated with various concentrations （0.1, 1, 10, 20, 60, 80, 100 μg/L） of rhHGF. Then, MTT method was used to detect the proliferation of treated melanocytes; also, tyrosinase activity was measured. Morphological changes of melanocytes treated with rhHGF at the concentration of 60 μg/L were observed by an inverted microscope, cell apoptosis and cycle by flow cytometry. Results The proliferation of melanocytes was significantly enhanced by the Ireatment with rhHGF at the concentrations of 20, 60 and 80 μg/L （P 〈 0.01 ）, and the activity of tyrosinase by the treatment with rhHGF at the concentrations from 0.1 to 100 μg/L （P〈 0.01 or 0.05）. The treatment with rhHGF at the concentration of 60μg/L for 48 hours resulted in the increase of the cell number, size and dendrite amount of melanocytes, acceleration of the transformation of these cells from G0/G1 phase to S phase, and inhibition of apoptosis of the cells. Conclusion rhI-IGF can enhance the biologic activity of cultured human melanocytes/n vitro.