低分子量可溶性壳寡糖对MC3T3-E1成骨细胞增殖和分化的影响

Effects of water soluble chitosan on the proliferation and differentiation of osteoblast-MC3T3-E1

目的探讨低分子可溶性壳寡糖对小鼠成骨细胞MC3T3-E1增殖和分化作用的影响及机制。方法培养小鼠成骨细胞MC3T3-E1,将细胞分为高、中、低浓度壳寡糖组(1 000、500和250μg.mL-1)、过氧化氢(H2O2)模型组、壳寡糖预处理过氧化氢组和对照组(只加同体积的培养液);采用MTT法、流式细胞术和碱性磷酸酶(ALP)的测定观察不同浓度的壳寡糖对成骨细胞的增殖和分化作用,观察壳寡糖对氧化损伤成骨细胞模型的作用.结果与结论低分子可溶性壳寡糖能显著提高小鼠成骨细胞MC3T3-E1的增殖和分化;降低细胞的凋亡率,使细胞ALP的活性提高;能减轻过氧化氢对细胞的损伤,提高细胞的增殖和分化。其作用机制可能与壳寡糖的抗氧化作用有关。

Objective To study the effects of water-soluble chitosan on the proliferation and differentiation of osteoblast-MC3T3-E1.Methods MC3T3-E1 osteoblasts were randomly divided in six groups：three test groups including high,medium and low-dose water soluble chitosan group（1 000,500,250 μg·mL-1）,H2O2 model group,H2O2 model group pretreatment with chitosan and control group;The cell viability was determined by MTT method and the indiex ofALP was tested by biochemical method;The rate of apoptosis of MC3T3-E1 was tested using flow cytometry（FCM）.Results and Conclusion The results showed that water soluble chitosan could significantly increase the viability of MC3T3-E1 and decreased the rate of apoptosis.The ALP concentrations in serum of the water soluble chitosan groups were higher signifcantly than that of the control group.Compared with the model group,the cell proliferation and differentiation of chitosan pretreatment groups increased.These results indicated that water soluble chitosan could promote the proliferation and differentiation of osteoblast-MC3T3-E1.The mechanisms of the effects might be related to the elimination of oxygen free radical in cells.