小檗碱对喉癌细胞克隆、转移及Toll样受体4蛋白的作用  

Effects of Berberine on the Cloning,Metastasis,and TLR4 Protein of Laryngeal Carcinoma Cells

作　　者：李娜[1] 张颖 李芳园 焦培英 张义[1] 王文娟 LI Na;ZHANG Ying;LI Fangyuan;JIAO Peiying;ZHANG Yi;WANG Wenjuan(Harleeson International Peace Hospital,Hengshui 053000,China;Hebei University of Engineering,Handan 056038,China)

机构地区：[1]哈励逊国际和平医院,衡水053000 [2]河北工程大学,邯郸056038

出　　处：《世界中医药》2022年第20期2867-2872,共6页World Chinese Medicine

基　　金：河北省医学重点课题计划项目(20181582)。

摘　　要：目的:研究不同浓度小檗碱对喉癌细胞克隆、转移及Toll样受体4蛋白的影响与表达。方法:将喉癌细胞分为空白对照组、5μmol/L小檗碱干预组、10μmol/L小檗碱干预组、20μmol/L小檗碱干预组。蛋白质印迹法检测Toll样受体4、Toll样受体2蛋白水平;噻唑蓝(MTT)法检测细胞增殖;划痕实验检测细胞迁移情况;流式法检测细胞凋亡。结果:在24 h时,空白对照组的细胞克隆形成能力与5μmol/L小檗碱干预组相似(P>0.05),空白对照组的细胞克隆能力较10μmol/L小檗碱干预组、20μmol/L小檗碱干预组高(P<0.05);在48 h和72 h时,与空白对照组比较,其他加入小檗碱进行干预的3组细胞克隆形成率均有明显下降(P<0.05)。且20μmol/L小檗碱干预组的克隆形成率明显低于10μmol/L小檗碱干预组、5μmol/L小檗碱干预组(P<0.05);与空白对照组比较,5μmol/L小檗碱干预组、10 mol/L小檗碱干预组、20μmol/L小檗碱干预组细胞中Toll样受体2及Toll样受体4蛋白表达降低(P<0.05),与5μmol/L小檗碱干预组、10μmol/L小檗碱干预组比较,20μmol/L小檗碱干预组Toll样受体2及Toll样受体4蛋白下降(P<0.05);与空白对照组比较,5μmol/L小檗碱干预组、10μmol/L小檗碱干预组、L4组细胞增殖率、划痕距离降低,凋亡率升高(P<0.01);与5μmol/L小檗碱干预组、10μmol/L小檗碱干预组比较,20μmol/L小檗碱干预组细胞的增殖率、划痕距离降低,凋亡率升高(P<0.05)。结论:不同浓度小檗碱均可降低喉癌细胞克隆、增殖及迁移能力,其作用机制可能通过抑制Toll样受体2、Toll样受体4蛋白活性从而促进喉癌细胞凋亡。Objective:To study the effects of different concentrations of berberine on the cloning,metastasis,and Toll-like receptor 4(TLR4)protein of laryngeal carcinoma cells.Methods:The laryngeal carcinoma cells were divided into a blank control group(K1 group),a 5μmol/L berberine intervention group(K2 group),a 10μmol/L berberine intervention group(K3 group),and a 20μmol/L berberine intervention group(K4 group).Western blotting method was used to detect TLR4 and TLR2 protein levels,and the methyl thiazolyl tetrazolium(MTT)assay was used to detect cell proliferation.The scratch test was used to detect cell migration,and the flow cytometry was used to detect cell apoptosis.Results:At 24 h,the cell clone formation ability of the K1 group was similar to the K2 group(P>0.05),and the cell clone ability of the K1 group was higher than that of the K3 and K4 groups(P<0.05).At 48 h and 72 h,compared with the K1 group,the other three groups had a significant decrease in the colony formation rate(P<0.05).Moreover,the clone formation rate of the K4 group was significantly lower than that of the K3 and K2 groups(P<0.05).Compared with the K1 group,the K2,K3,and K4 groups decreased TLR2 and TLR4 proteins(P<0.05).Compared with the K2 and K3 groups,the K4 group decreased TLR2 and TLR4 proteins(P<0.05).Compared with the K1 group,the K2,K3,and L4 groups decreased cell proliferation rate and scratch distance,and increased apoptosis rate(P<0.01).As compared with the K2 group and K3 group,the cell proliferation rate and scratch distance of the K4 group decreased,and the apoptosis rate increased(P<0.05).Conclusion:Different concentrations of berberine can reduce the cloning,proliferation,and migration ability of laryngeal carcinoma cells,and the mechanism may be related to the inhibition of the activity of TLR2 and TLR4 proteins,thereby promoting the apoptosis of laryngeal carcinoma cells.

关 键 词：小檗碱 喉癌细胞 克隆 增殖 划痕试验 凋亡 TOLL样受体4 

分 类 号：R284[医药卫生—中药学] R273[医药卫生—中医学]