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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-β_1,TGF-β_1)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-β_1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 this paper, MTS [3- (4,5-dimethylthiazol-2-yl) -5- (3-carboxymethoxyphenyl) -2 (4-sulfophenyl) -2H-tetrazolium] The proliferation of GES-1 cells was evaluated by capillary electrophoresis. The monolayer fused GES-1 cells were scratched with capillary to simulate gastric mucosal injury. The migration of GES-1 cells in wound area was observed and calculated after GFP treatment And repair the damaged area. The levels of epidermal growth factor (EGF), transforming growth factor-β (TGF-β_1, TGF-β_1) and clover leaf were determined by enzyme-linked immunosorbent assay (ELISA) (TFF2), and the changes of m RNA were detected by real-time quantitative PCR (RT-PCR). The experimental results show that GFP can promote the proliferation of GES-1 cells by up-regulating EGF and TFF2 and down-regulating the expression of TGF-β 1, and promote the migration of GES-1 cells to the wound area, which can completely cover the wound area and repair the gastric mucosa.