白及多糖联合5-氟尿嘧啶对人肝癌HepG2细胞凋亡的影响  

Effect of Bletilla Striata Polysaccharide Combined with 5-Fluorouracil on Apoptosis of Human Hepatoma HepG2 Cells

作　　者：温艳 张强 曹涵博 WEN Yan;ZHANG Qiang;CAO Hanbo(Shaanxi Institute for Food and Drug,Xi'an,Shaanxi,China 710065;Shaanxi Provincial Drug and Vaccine Inspection Center,Xi'an,Shaanxi,China 710065)

机构地区：[1]陕西省食品药品检验研究院,陕西西安710065 [2]陕西省药品疫苗检查中心,陕西西安710065

出　　处：《中国药业》2022年第24期47-50,共4页China Pharmaceuticals

摘　　要：目的 探讨白及多糖(BSP)联合5-氟尿嘧啶(5-FU)对人肝癌HepG2细胞凋亡的影响。方法 以0,50,100,200,400,800μg/mL BSP分别培养细胞24,48,72 h,采用CCK-8法,测定细胞活力,确定BSP最优培养质量浓度。另设对照组(等体积生理盐水)、BSP组(400μg/mL)、5-FU组(0.02 mol/L)、BSP+5–FU组(400μg/mL+0.02 mol/L),培养细胞48 h,测定细胞活力;采用流式细胞术检测细胞凋亡情况;采用荧光定量聚合酶链式反应法检测半胱氨酸蛋白水解酶8(Caspase-8)、B淋巴细胞瘤2(Bcl-2)和Bcl-2相关X蛋白(BAX)mRNA水平;采用Western blot法检测Caspase-8、Bcl-2和BAX蛋白表达水平。结果 与0μg/mL比较,400μg/mL BSP作用48 h后细胞活力开始显著降低(P <0.05)。与5-FU组及BSP组比较,BSP+5-FU组细胞活力均显著减低(P <0.05);细胞凋亡率均显著升高(P <0.05);Caspase-8和BAX mRNA和蛋白表达水平均显著升高(P <0.05),Bcl-2 mRNA和蛋白表达水平均有所降低。结论 BSP联合5-FU可诱导HepG2细胞凋亡,其机制可能与Caspase-8介导的细胞凋亡有关。Objective To investigate the effect of Bletilla striata polysaccharide(BSP) combined with 5-fluorouracil(5-FU) on the apoptosis of human hepatoma HepG2 cells.Methods The cells were cultured at 0,50,100,200,400,800 μg/mL of BSP for24,48 and 72 h,respectively. The cell viability was measured by CCK-8 method to determine the optimal concentration of BSP.The control group(equal volume of distilled water),BSP group(400 μg/mL),5-FU group(0. 02 mol/L),and BSP + 5-FU group(400 μg/mL + 0. 02 mol/L) were set up,and the cells in those groups were cultured for 48 h to measure the cell viability.The apoptosis of cells was detected by flow cytometry. The expression levels of Caspase-8,B lymphocytoma 2(Bcl-2) and Bcl-2related X protein(BAX) mRNA were detected by fluorescence quantitative polymerase chain reaction. The expression levels of Caspase-8,Bcl-2 and BAX proteins were detected by Western blot.Results Compared with treatment with 0 μg/mL of BSP,the cell viability began to decrease significantly after 48 h of treatment with 400 μg/mL of BSP(P < 0. 05). Compared with those in the 5-FU group and BSP group,the cell viability in the BSP + 5-FU group decreased significantly(P < 0. 05),the apoptosis rate in the BSP + 5-FU group significantly increased(P < 0. 01),the levels of Caspase-8 and BAX mRNA and protein increased significantly(P < 0. 05),while the level of Bcl-2 mRNA and protein decreased.Conclusion BSP combined with 5-FU can induce apoptosis of HepG2 cells,which may be related to Caspase-8 pathway.

关 键 词：白及多糖 5-氟尿嘧啶 人肝癌HEPG2细胞 细胞凋亡 CASPASE-8 

分 类 号：R932[医药卫生—生药学]