人参皂苷Rg1对人喉癌Hep2细胞株增殖及一氧化氮合酶通路的影响  

Significanee of ginsenoside Rg1 in laryngeal cancer Hep2 cells proliferation and eNOS/iNOS pathways

作　　者：李建涛 孔婷婷 孙毓晗 米光煕 王乐秋 Li Jiantao;Kong Tingting;Sun Yuhan;Mi Guangxi;Wang Leqiu(Department of Otolaryngology,the Affiliated Hongqi Hospital of Mudanjiang Medical University,Mudanjiang 157000,China)

机构地区：[1]牡丹江医学院附属红旗医院耳鼻咽喉科,黑龙江157000

出　　处：《中国实用医刊》2020年第3期10-13,共4页Chinese Journal of Practical Medicine

基　　金：黑龙江省属高等学校科研攻关项目(2018-KYYWFMY-0054)。

摘　　要：目的探讨人参皂苷Rg1对人喉癌Hep2细胞株增殖及一氧化氮合酶(eNOS/iNOS)通路的影响。方法利用人参皂苷Rg1(药物浓度分别为40、60、80μmol/L)处理喉癌Hep2细胞24、48、72 h,为人参皂苷组,并设正常培养的对照组。采用四甲基偶氮唑盐微量酶反应比色法检测各组细胞的增殖活力;采用流式细胞仪分析各组细胞周期处于各时相的比例;采用逆转录聚合酶链式反应(RT-PCR)法检测细胞eNOS及iNOS的表达水平。结果Rg1对Hep2细胞的增殖具有明显的抑制作用,且呈时间和剂量依赖性,80μmol/L为最适作用浓度。与对照组相比,经80μmol/L的Rg1作用24、48、72 h后,Hep2细胞的凋亡率逐渐增加(P<0.01);S期和G2/M期细胞比例逐渐下降,G0/G1期细胞比例逐渐升高(P<0.01)。与对照组相比,人参皂苷组细胞iNOS含量降低,而NO、eNOS含量上升。人参皂苷组Hep2细胞株中iNOS的mRNA表达高于对照组,eNOS的mRNA表达低于对照组,这与细胞分光光度计法测定结果相一致。结论人参皂苷Rg1可以抑制人喉癌细胞Hep2细胞增殖,其作用机制可能为人参皂苷Rg1的抗氧化机制增加eNOS表达,抑制iNOS表达,增加NO活性。Objective To investigate the significance of ginsenoside Rg1 in laryngeal cancer Hep2 cells proliferation and endothelial nitric oxide/inducible nitric-oxide synthse (eNOS/iNOS) pathways.Methods In the study, laryngeal cancer Hep2 cells were calculated by ginsenoside Rg1, with drug concentrations as 40 μmol/L, 60 μmol/L, 80 μmol/L , respectively, and cells in different concentrations were all cultured for 24, 48, 72 h as ginsenoside Rg1 group;and a normal training was set as the control group. Methyl thiazolyl tetrazolium method was used to test cell proliferation activity of every group. Flowcytometry analysis was used to analyze the proportion of each cell cycle in each phase. Transcription-polymerase chain reaction method was used to detect cell eNOS and iNOS expression level.Results Rg1 had inhibitory effects on Hep2 cells proliferation, showing time and dose dependent, with 80 μmol/L concentration for the optimum effect. Compared with control group, after been calculated by 80 μmol/L Rg1 for 24, 48, 72 h, Hep2 cells apoptosis rate increased gradually (P<0.01);S phase and G2/M phase cells proportion gradually declined, while G0/G1 phase cell proportion increased (P<0.01). Compared with control group, iNOS levels of ginsenoside Rg1 group decreased, but the content of NO and eNOS was higher. Compared with control group, ginsenoside Rg1 group had higher expression of iNOS mRNA in Hep2 cell lines, and lower eNOS mRNA expression, which was in line with results of spectrophotometric determination.Conclusions Ginsenoside Rg1 can inhibit human laryngeal cancer cells Hep2 cells proliferation, and its mechanism may be related to antioxidant effect of ginsenoside Rg1, by which the eNOS expression is promoted, iNOS expression is inhabited, and the NO activity is improved.

关 键 词：人参皂苷RG1 人喉癌Hep2细胞 一氧化氮合酶 一氧化氮 

分 类 号：R28[医药卫生—中药学]