三物白散对胃癌细胞自噬相关蛋白Beclin1、LC3、p62表达的影响  

The Effect of Sanwubai Powder(三物白散)on the Expression of Autophagy Related Proteins Beclin1,LC3 and p62 in Gastric Carcinoma Cells

作　　者：袁宇晴 龙丹 谢超群 魏堰翀 刘旭东 朱莹 邹君君[3] YUAN Yuqing;LONG Dan;XIE Chaoqun;WEI Yanchong;LIU Xudong;ZHUYing;ZOU Junjun(Hunan University of Chinese Medicine,Changsha Hunan 410208,China;The First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha Hunan 410007,China;The Second Affiliated Hospital of Hunan University of Chinese Medicine,Changsha Hunan 410005,China)

机构地区：[1]湖南中医药大学,湖南长沙410208 [2]湖南中医药大学第一附属医院,湖南长沙410007 [3]湖南中医药大学第二附属医院,湖南长沙410005

出　　处：《中医药导报》2024年第3期30-36,共7页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：湖南省科技厅青年基金项目(2021JJ40415)。

摘　　要：目的:探讨三物白散对胃癌细胞自噬相关蛋白Beclin1、LC3、p62表达的影响,探究三物白散抗胃癌的作用机制。方法:将28只SPF级雄性大鼠随机分为4组,每组7只,分别予以0.03125、0.0625、0.1250 g/(kg·d)三物白散及相同体积生理盐水灌胃,经腹主动脉采血制备含药血清。体外培养AGS细胞,设置空白对照组(空白组)、三物白散低剂量含药血清组(低剂量组)、三物白散中剂量含药血清组(中剂量组)、三物白散高剂量含药血清组(高剂量组)。采用CCK-8法检测不同浓度(5%、10%、15%、20%)三物白散含药血清对AGS细胞活性的影响;倒置显微镜下观察三物白散含药血清对AGS细胞形态的影响;RT-qPCR法检测Beclin1 mRNA、LC3 mRNA、p62 mRNA表达;Western blotting法检测Beclin1、LC3、p62蛋白表达。结果:与空白组比较,5%、10%、15%、20%三物白散高剂量含药血清可呈浓度-时间依赖性抑制AGS细胞活性;随着含药血清浓度升高,细胞形态逐渐不规则,细胞数量逐渐减少;中、高剂量组Beclin1 mRNA、LC3-ⅠmRNA、LC3-ⅡmRNA相对表达量均高于空白组,p62 mRNA相对表达量低于空白组(P<0.01);低剂量组LC3-ⅠmRNA、LC3-ⅡmRNA相对表达量高于空白组,p62 mRNA相对表达量低于空白组(P<0.01)。高剂量组Beclin1蛋白相对表达量及LC3-Ⅱ/LC3-Ⅰ均高于空白组,p62蛋白相对表达量低于空白组(P<0.01);中剂量组Beclin1蛋白相对表达量及LC3-Ⅱ/LC3-Ⅰ高于空白组(P<0.05或P<0.01),p62蛋白相对表达量低于空白组(P<0.01)。结论:三物白散对胃癌细胞的增殖有抑制作用,上调Beclin1、LC3表达、下调p62的表达、诱导胃癌细胞自噬可能是其抗胃癌作用机制之一。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 expression 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-ⅠmRNA and LC3-ⅡmRNA 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-ⅠmRNA and LC3-ⅡmRNA 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

关 键 词：胃癌 三物白散 自噬 BECLIN1 LC3 P62 

分 类 号：R285.5[医药卫生—中药学]