灰树花多糖对体外胃粘膜创伤的修复作用

In vivo protective and repaired effects of polysaccharide from Grifola frondosa on gastric mucosa trauma

本文采用MTS[3-（4,5-dimethylthiazol-2-yl）-5-（3-carboxymethoxyphenyl）-2（4-sulfophenyl）-2H-tetrazolium]法检测灰树花多糖（GFP）对人胃正常粘膜上皮细胞（human gastric epithelial cell,GES-1）细胞增殖影响;使用毛细管对单层融合的GES-1细胞进行划痕,模拟胃粘膜创伤,观察并计算GFP作用后GES-1细胞创伤区域迁移及修复损伤的面积;ELISA（enzyme-linked immunosorbent assay）方法测定培养基上清液中表皮生长因子（epidermal growth factor,EGF）、转移生长因子（transforming growth factor-β1,TGF-β1）和三叶因子2（trefoil factors 2,TFF2）的含量变化;荧光定量PCR（RT-PCR）检测其m RNA的变化。实验结果表明：GFP通过上调EGF、TFF2,下调TGF-β1表达而促进GES-1细胞增殖,促进其向创伤区域迁移,可以完全覆盖创伤区域达到修复胃粘膜作用。

The effect of polysaccharide of Grifola frondosa（GFP） on the proliferation of gastric epithelial cell GES-1 was detected by 3-（4,5-dimethylthiazol-2-yl）-5-（3-carboxymethoxyphenyl）-2-（4-sulfophenyl）-2H-tetrazolium（MTS）. The monolayer of the confluent GES-1 cells was cut by capillary to simulate the gastric mucosa injury, then the migration of the GES-1 cells and the repair of the wounded area were evaluated. The contents of epidermal growth factor（EGF）, transforming growth factor-β1（TGF-β1） and trefoil factors 2（TFF2） in the supernatant of culture medium were determined by enzyme-linked immunosorbentassay（ELISA）, and the change of expression level of rn RNA was detected by RT-PCR. The results indicated that GFP promotes the GES-1 cell proliferation and migration to the wounded area to repair the gastric mucosa membrane through up-regulating the expression of EGF and TFF2, and down-regulating the expression of TGF-β1.