灰树花多糖对体外胃粘膜创伤的修复作用

本文采用MTS3-(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mRNA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.

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

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 immunosorbent assay (ELISA), and the change of expression level of rnRNA 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.