The Effect of Sanwubai Powder(三物白散)on the Expression of Autophagy Related Proteins Beclin1,LC3 and p62 in Gastric Carcinoma Cells
Objective:By studying the effect of Sanwubai Powder on the expression of autophagy related proteins Beclin1,LC3,and p62 in gastric carcinoma cells,we aim to explore the partial mechanism of Sanwubai Powder's anti gastric carcinoma effect.Methods:Totally 28 SPF grade male rats were randomly divided into four groups,seven rats in each group,and the four groups were given 0.03125,0.0625,0.125 g/(kg·d)Sanwubai Powder and the same volume of physiological saline by gavage.Blood was collected from the abdominal aorta to obtain drug-containing serum.In vitro cultivation of AGS cells,four groups were established,including blank control group(blank group),low dose medicated serum group(low dose group),medium dose medicated serum group(medium dose group)and high dose medicated serum group(high dose group).The CCK-8 method was used to detect the effect of medicated serum containing Sanwubai Powder at different concentrations(5％,10％,15％and 20％)on the activity of AGS cells.Inverted microscope was used to observe the effect of Sanwubai Powder medicated serum on the morphology of AGS cells.RT qPCR was used to detect the expres-sion of Beclin1 mRNA,LC3 mRNA and p62 mRNA.Western blotting was used to detect the expression of Beclin1,LC3 and p62 proteins.Results:Compared with the blank group,high-dose serum containing 5％,10％,15％,and 20％Sanwubai Powder can inhibit AGS cell activity in a concentration time dependent manner.As the concentration of drug containing serum increases,the cell morphology gradually becomes irregular and the number of cells gradually decreases.The medium and high dose groups showed higher relative expression levels of Beclin1 mRNA,LC3-Ⅰ mRNA and LC3-Ⅱ mRNA than blank group,while lower relative expression levels of p62 mRNA than blank group(P＜0.01).The low dose group showed higher relative expression levels of LC3-Ⅰ mRNA and LC3-Ⅱ mRNA than blank group,while lower relative expression levels of p62 mRNA than blank group(P＜0.01).The highdose group showed higher relative expression level of Beclin1 protein and LC3-Ⅱ/LC3-Ⅰ than blank group,while lower relative expression level of p62 protein than blank group(P＜0.01).The medium dose group showed higher relative expression level of Beclin1 protein and LC3-Ⅱ/LC3-Ⅰthan blank group(P＜0.05 or P＜0.01),while lower relative expression level of p62 protein than blank group(P＜0.01).Conclusion:Sanwubai Powder can inhibit proliferation of gastric carcinoma cells,up-regulate the expression of Beclin1 and LC3,down-regulate the expression of p62,and induce autophagy in gastric carcinoma cells,which may be one of the mechanisms of its antigastric carcinoma effect.
万方数据
维普
