淫羊藿苷激活内质网应激对人结肠癌细胞的凋亡作用  

Effect of icariin activating endoplasmic reticulum stress on apoptosis of human colon cancer cells

作　　者：王璐 王杰[2,3] 陈腾[2,3] 殷佩浩[2,3] WANG Lu;WANG Jie;CHEN Teng;YIN Pei-hao(School of Medicine and Life Sciences,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,Sichuan Province,China;Department of General Surgery,Putuo Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 200062,China;Shanghai Putuo Center Clinical College,Anhui Medical University,Shanghai 200062,China)

机构地区：[1]成都中医药大学医学与生命科学学院,四川成都611137 [2]上海中医药大学附属普陀医院普外科,上海200062 [3]安徽医科大学上海普陀中心临床学院,上海200062

出　　处：《中国临床药理学杂志》2023年第21期3106-3110,共5页The Chinese Journal of Clinical Pharmacology

基　　金：国家科技部重点研发计划基金资助项目(2019YFC1316003);国家自然科学基金面上基金资助项目(81973625);上海市普陀区卫生健康系统临床特色专病建设基金资助项目(2019tszb01)。

摘　　要：目的基于内质网应激探讨淫羊藿苷(ICA)对人结肠癌细胞HCT116、SW620增殖、凋亡的影响。方法选取不同浓度的ICA处理HCT116、SW620细胞48 h,分别用平板细胞克隆实验、Annexin-V/PI流式凋亡检测法和蛋白质印迹(Western Blot)法检测ICA对HCT116、SW620细胞的增殖抑制作用、凋亡的影响和内质网应激相关蛋白表达的影响。将HCT116、SW620细胞分别分为对照组、ICA组、ICA+4-PBA组。对照组均用完全培养基培养48 h;ICA组分别用80、100μmol·L^(-1)ICA培养HCT116、SW62048 h;ICA+4-PBA组分别用80μmol·L^(-1)ICA+2 mmol·L^(-1)4-PBA、100μmol·L^(-1)ICA+2 mmol·L^(-1)4-PBA培养HCT116、SW62048 h;用蛋白质印迹法检测各组细胞中内质网应激相关蛋白的表达。结果ICA对HCT116细胞48 h IC50值为111.10μmol·L^(-1),对SW620细胞48 h IC50值为159.40μmol·L^(-1)。平板细胞克隆实验结果显示,随着ICA浓度的增加,HCT116、SW620细胞增殖减少(P<0.05)。流式细胞术显示,HCT116细胞48 h的0、55、110、220μmol·L^(-1)ICA组凋亡率分别为(6.70±3.19)%、(12.73±1.90)%、(31.71±3.30)%、(56.85±3.66)%,SW620细胞48 h的0、80、160、320μmol·L^(-1)ICA组凋亡率分别为(3.28±0.12)%、(10.27±0.77)%、(22.34±2.16)%、(50.35±2.17)%。Western Blot结果显示,随着ICA浓度的增加,HCT116、SW620细胞凋亡蛋白c-cas3、Bax表达增加,抗凋亡蛋白Bcl-2表达下调,内质网应激相关蛋白表达上调。结论淫羊藿苷对HCT116、SW620细胞具有抑制增殖、促凋亡的作用,其机制可能与上调内质网应激信号通路有关。Objective To investigate the effect of icariin(ICA)on proliferation and apoptosis of human colon cancer cells HCT116 and SW620 based on endoplasmic reticulum stress.Methods HCT116 and SW620 cells were treated with different concentrations of ICA for 48 h.The inhibitory effect of ICA on the proliferation and apoptosis of HCT116 and SW620 cells and the expression of endoplasmic reticulum stress-related proteins p-perk,perk,Bip,p-eif2αand eif2αwere detected by plate cell clone test,Annexin-V/PI flow cytometry and Western Blot,respectively.HCT116 and SW620 cells were divided into control group,ICA group and ICA+4-PBA group.The control group was cultured in complete medium for 48 h.In ICA group,HCT116 and SW 620 were cultured with 80 and 100μmol·L^(-1) ICA,and HCT116 and SW620 were cultured with 80μmol·L^(-1) ICA+2 mmol·L^(-1)4-PBA and 100μmol·L^(-1) ICA+2 mmol·L^(-1)4-PBA for 48 h,respectively.Western Blot was used to detect the expression of endoplasmic reticulum stress related proteins in each group.Results The 48 h IC50 values of ICA on HCT116 cells and SW620 cells were 111.10 and 159.40μmol·L^(-1),respectively.The results of plate cell clone assay showed that the proliferation of HCT116 and SW620 cells decreased with the increase of ICA concentration(P<0.05).Flow cytometry showed that the apoptosis rates of HCT116 cells in 0,55,110,220μmol·L^(-1) ICA groups were(6.70±3.19)%,(12.73±1.90)%,(31.71±3.30)%,(56.85±3.66)%and in SW620 cells of 0,80,160,320μmol·L^(-1) ICA groups were(3.28±0.12)%,(10.27±0.77)%,(22.34±2.16)%,(50.35±2.17)%,respectively.Western Blot results showed that with the increase of ICA concentration,the expression of apoptotic protein c-cas3 and Bax in HCT116 and SW620 cells increased,the expression of anti-apoptosis protein Bcl-2 decreased,and the expression of endoplasmic reticulum stress related proteins were up-regulated.Conclusion Icariin can inhibit proliferation and promote apoptosis of HCT116 and SW620 cells,and its mechanism may be related to the up-regulation of endop

关 键 词：淫羊藿苷 人结肠癌细胞 细胞凋亡 内质网应激 

分 类 号：R28[医药卫生—中药学]