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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-1H2O2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。
Objective To investigate the effect of Ganoderma lucidum polysaccharides on the proliferation, migration and differentiation of IEC-6 cells in vitro in rat intestinal crypt cells IEC-6 cells cultured in vitro. Methods After different concentrations of Ganoderma lucidum polysaccharides were exposed to normal IEC-6 cells for 44 h, the proliferation of cells was measured by thiazolyl blue (MTT) method. Ganoderma lucidum polysaccharides were pretreated with IEC-6 cells for 46 h, and IEC-6 cells were stimulated with 500 μmol·L-1H2O2 for 40 min. The proliferation of cells was measured by MTT assay. The IEC-6 monolayer model of intestinal epithelial cell damage was used to detect the effect of Ganoderma lucidum polysaccharides on the migration of IEC-6 cells, and the content of TGF-β in culture supernatant was detected by ELISA to explore the migration pathway. The cells were subjected to HE staining to observe the differentiation of IEC-6 cells. RESULTS: Ganoderma lucidum polysaccharides promoted proliferation of IEC-6 cells after normal and H2O2 injury, and showed a dose-effect relationship; 10 mg·L-1 Ganoderma lucidum polysaccharides could promote the migration of damaged IEC-6 cells, but the cell supernatant No effect on the content of TGFβ. Under the action of 10 mg·L-1 Ganoderma lucidum polysaccharide, the degree of differentiation of IEC-6 cells increased. Conclusion Ganoderma lucidum polysaccharide can promote the proliferation, migration and differentiation of IEC-6 cells. Its role in promoting migration is independent of the TGFβ pathway.