芍药苷通过抑制Notch-1信号通路影响肝癌细胞增殖、侵袭和凋亡的机制研究  

Effect of paeoniflorin on hepatocellular carcinoma(HCC)cell proliferation,invasion,and apoptosis by inhibiting Notch-1 signaling pathway

作　　者：夏亮[1] 谢齐贵[1] 陈湛蕾 XIA Liang;XIE Qi-gui;CHEN Zhan-lei(Department of Gastroenterology,Tongde Hospital of Zhejiang Province,Hangzhou,Zhejiang 310012,China)

机构地区：[1]浙江省立同德医院消化科,杭州310012

出　　处：《浙江中西医结合杂志》2022年第6期520-523,共4页Zhejiang Journal of Integrated Traditional Chinese and Western Medicine

摘　　要：目的探讨芍药苷对肝癌细胞增殖、侵袭和凋亡的影响及其机制。方法使用不同浓度(0、25、50、75、100、150μmol/L)芍药苷处理人肝癌细胞系HepG2,采用CCK-8法检测细胞增殖活性。根据芍药苷浓度将HepG2细胞株分为四组:空白对照组(0μmol/L)、芍药苷低剂量组(25μmol/L)、芍药苷中剂量组(50μmol/L)和芍药苷高剂量组(75μmol/L)。采用流式细胞仪检测细胞凋亡情况,Transwell法细胞侵袭迁移能力,Western blot法测定Notch-1和Hes-1蛋白水平。结果使用不同浓度(0、25、50、75、100、150μmol/L)芍药苷处理HepG2细胞24 h后,细胞增殖活性呈剂量依赖性降低[(83.34±8.06)%、(74.46±7.34)%、(63.59±2.51)%、(55.93±4.05)%、(39.99±6.06)%比100%,P<0.05]。当不同浓度(0、25、50、75μmol/L)芍药苷处理HepG2细胞24 h后,细胞穿膜数逐渐降低[(64.33±5.13)个、(49.67±3.51)个、(30.00±2.00)个比(85.33±5.51)个,P<0.05],细胞凋亡率逐渐升高[(13.37±2.57)%、(22.64±2.19)%、(28.53±1.37)%比(2.43±0.93)%,P<0.05],Notch-1蛋白表达水平逐渐降低[(0.75±0.05)、(0.55±0.06)、(0.39±0.04)比(1.13±0.15),P<0.05],Hes-1蛋白表达水平也逐渐降低[(0.55±0.07)、(0.38±0.04)、(0.17±0.03)比(0.80±0.07),P<0.05]。结论芍药苷呈剂量依赖性抑制肝癌细胞HepG2增殖、侵袭,并可促进细胞凋亡,其机制可能与抑制Notch-1信号通路相关。Objective To assess the effect of paeoniflorin on hepatocellular carcinoma(HCC)cell proliferation,invasion,and apoptosis and then explore the underlying molecular events.Methods Human HCC HepG2 cells were cultured and treated with different doses of paeoniflorin(0,25,50,75,100 and 150μmol/L)and then subjected to cell viability CCK-8 assay.After that,cells were divided into blank control(0μmol/L)and low(25μmol/L),medium(50μmol/L),and high-dose paeoniflorin group(75μmol/L).Flow cytometry assay was performed to detect cell apoptosis and transwell assay was used to assess cell invasion ability.Western blot assay was performed to measure the expression levels of Notch-1 and Hes-1 protein.Results Different doses of paeoniflorin(0,25,50,75,100 and 150μmol/L)reduced HepG2 cell viability after 24 h treatment in a dose-dependent manner(83.34±8.06,74.46±7.34,63.59±2.51,55.93±4.05 and 39.99±6.06 vs.100%;P<0.05).These different paeoniflorin doses(0,25,50 and 75μmol/L)for 24 h treatment also reduced tumor cell invasion capacity(64.33±5.13,49.67±3.51 and 30.00±2.00 vs.85.33±5.5;P<0.05),whereas rates of tumor cell apoptosis were upregulated(13.37±2.57,22.64±2.19 and 28.53±1.37 vs.2.43±0.93%;P<0.05).At the gene level,these treatments downregulated levels of Notch-1(0.75±0.05,0.55±0.06 and 0.39±0.04 vs.1.13±0.15;P<0.05)and Hes-1 proteins(0.55±0.07,0.38±0.04 and 0.17±0.03 vs.0.80±0.07;P<0.05).Conclusion Paeoniflorin treatment was able to dose-dependently inhibit HCC cell proliferation and invasion,but promoted them to undergo apoptosis in vitro.The underlying molecular events may be due to its inhibition of the Notch-1 signaling.

关 键 词：芍药苷 肝癌 增殖 侵袭 凋亡 Notch-1信号通路 

分 类 号：R285[医药卫生—中药学]