灵芝多糖对IEC-6细胞促增殖、迁移与分化作用

 目的通过灵芝多糖作用于体外培养的大鼠小肠隐窝细胞株IEC-6细胞,观察灵芝多糖对IEC-6细胞增殖、迁移与分化的影响。方法不同浓度的灵芝多糖作用于正常IEC-6细胞44h后,用噻唑蓝(MTT)法测细胞的增殖。灵芝多糖预作用IEC-6细胞46h后,用500μmol·L^{-12O₂刺激IEC-6细胞40min,以MTT法测细胞的增殖变化。采用IEC-6单层肠上皮细胞损伤重建模型,检测灵芝多糖对IEC-6细胞迁移的影响,并用ELISA方法检测培养细胞上清中TGF-β的含量以探讨迁移途径。对细胞进行HE染色来观察IEC-6细胞的分化情况。结果灵芝多糖对正常及受H₂O₂损伤后的IEC-6细胞均有促增殖作用,并呈量效关系;10mg·L-1灵芝多糖可促进受损IEC-6细胞的迁移,但对细胞上清中TGFβ含量无影响。在10mg·L-1灵芝多糖作用下,IEC-6细胞分化程度增高。结论灵芝多糖可以促进IEC-6细胞增殖、迁移与分化。其促迁移作用是非依赖TGFβ途径的。}

Ganoderma Lucidum Polysaccharides Accelerated IEC-6 Cells Proliferation,Migration and Differentiation

OBJECTIVE To observe the effects of Ganoderma lucidum polysaccharides (Gl-PS) in different concentrations on the proliferation,wound restitution and differentiation of rat intestinal epithelial cell line IEC-6. METHODS IEC-6 cells were treated with different concentrations of Gl-PS for 44 h. The cells proliferation was estimated by MTT assay. IEC-6 cells were previously treated with different concentrations of Gl-PS for 46 h,then the cells were subjected to 500 μmol·L^-1 H₂O₂ stress for 40 min. The cells proliferation was also estimated by MTT assay. IEC-6 cells restitution was assessed using an in vitro monolayer wounding model. Transforming growth factor beta (TGF β) level of the cells culture supernatants were determined by ELISA. The cells differentiation was observed under microscope after HE stainning. RESULTS Compared with normal control,Gl-PS promoted cells proliferation in a dose-dependence manner. Pretreated with Gl-PS,IEC-6 cells growth inhibition was ameliorated under H₂O₂ stress. Gl-PS caused a significant dose-dependent acceleration of intestinal epithelial cell migration but had no effect on TGF β expression. Gl-PS induced the cell differentiation.CONCLUSION Gl-PS accelerated IEC-6 cells proliferation,migration and differentiation. The effect on IEC-6 cells migration was independent on TGF β route.