蒲公英赛醇通过调节AMPK通路保护胃黏膜细胞的作用和机制

The role and mechanism of taraxerol in protecting gastric epithelial cells by regulating AMPK pathway

目的通过体外实验研究,探讨AMPK信号通路在蒲公英赛醇保护乙醇损伤的胃黏膜细胞的作用。方法以乙醇刺激胃黏膜细胞为模型,蒲公英赛醇进行干预。采用CCK-8法检测乙醇刺激胃黏膜细胞状态下蒲公英赛醇对胃黏膜细胞存活率的影响,生化法检测各组胃黏膜细胞GSH-Px、SOD和MDA含量变化。流式细胞术检测各组胃黏膜细胞周期变化。Western blotting检测各组胃黏膜细胞AMPK、CaMKK和LKB1蛋白表达水平。结果蒲公英赛醇可浓度依赖性地改善乙醇刺激胃黏膜细胞的活性,胃黏膜细胞被乙醇损伤后的SOD和GSH-Px活性显著下降,MDA含量的明显升高;细胞增殖明显降低(P<0.05),G 1期细胞下降,S期和G 2期细胞上升;同时下调胃黏膜损伤细胞AMPK、CaMKK、LKB1蛋白表达水平。而蒲公英赛醇治疗后胃黏膜细胞内SOD、GSH-Px活性明显升高(P<0.01),MDA含量显著下降(P<0.01);细胞增殖明显增强(P<0.05),G 1期细胞上升,S期和G 2细胞下降(P<0.05)。并浓度依赖性地上调乙醇刺激下胃黏膜细胞中AMPK、CaMKK、LKB1蛋白表达水平。结论蒲公英赛醇可有效活化AMPK通路,从而保护受损的胃黏膜细胞。

Objective To investigate the protective effect of taraxerol on ethanol-injured gastric mucosal cells by AMPK signaling pathway by in vitro experimental study.Methods With ethanol to stimulate gastric mucosal cells as a model,taraxerol decoction to intervenet.The effect of taraxerol on the survival rate of gastric mucosa cells under the condition of ethanol stimulation was detected by CCK-8.The content of GSH-Px,SOD and MDA in gastric mucosa cells were measured by biochemical method.The cell cycle of gastric mucosa was detected by flow cytometry.Western blotting was used to detect the expression of AMPK,CaMKK and LKB1 protein.Results The activities of SOD and GSH-Px in gastric mucosal cells were significantly decreased and the content of MDA was significantly increased after the gastric mucosal cells were injured by ethanol.Cell proliferation was significantly decreased(P<0.05),cells in G 1 phase decreased,and cells in S phase and G 2 phase increased.The expression of AMPK,CaMKK and LKB1 in gastric mucosal injury cells was also down-regulated.The activities of SOD and GSH-Px were significantly increased(P<0.01),while the content of MDA was significantly decreased(P<0.01).The cell proliferation was significantly enhanced(P<0.05),the cells in G 1 phase were increased,and the cells in S phase and G 2 phase were decreased(P<0.05).The expression levels of AMPK,CaMKK and LKB1 in gastric mucosal cells were up-regulated by ethanol in a concentration-dependent manner.Conclusion Taraxerol can effectively activate the AMPK pathway,thereby protecting the damaged gastric mucosa cells.