萱草花总黄酮含药血清对肝癌 HepG2细胞增殖和凋亡的影响

摘要

目的：探讨萱草花总黄酮（HCBF）含药血清对人肝癌 HepG2细胞增殖和凋亡的影响，阐明其作用机制。方法：HepG2细胞分为空白对照组及低（900 mg·kg－1）、中（1800 mg·kg－1）和高（2700 mg·kg－1）剂量 HCBF 组。采用 MTT 法检测 HepG2细胞生长抑制率，倒置显微镜下观察 HepG2细胞形态表现， AnnexinⅤ-PI双染和流式细胞术检测细胞凋亡率，酶联免疫吸附法检测细胞中凋亡蛋白 Bax、bcl-2和 Caspase-3表达水平。结果：与空白对照组比较，中和高剂量 HCBF 组 HepG2细胞生长抑制率明显升高（P ＜0.05或 P ＜0.01）。倒置显微镜下观察，HCBF 组 HepG2细胞体积缩小，形状不规则，细胞核呈浓缩和破碎状态。与空白对照组比较，中和高剂量 HCBF 组 HepG2细胞凋亡率升高（P ＜0.05或 P ＜0.01），bcl-2蛋白表达水平明显降低（P ＜0.05或 P ＜0.01），Bax 蛋白表达水平明显升高（P ＜0.05或 P ＜0.01）；高剂量 HCBF 组 Caspase-3蛋白表达水平明显升高（P ＜0.05）。结论：HCBF 含药血清可以诱导 HepG2细胞凋亡，其机制可能与线粒体途经有关联。%Objective:To explore the effects of serum containing hemerocallis citrine baroni flavonids (HCBF)on the proliferation and apoptosis of human hepatoma HepG2 cells,and to clarify their mechanisms.Methods:The HepG2 cells were divided into blank control group, low (900 mg · kg-1 ), middle (1 800 mg · kg-1 )and high (2 700 mg·kg-1 )doses of serum containing HCBF groups.The inhibitory rates of growth of HepG2 cells were measured with MTT assay;inverted microscope was used to observe the morphologic changes;Annexin Ⅴ-PI double staining and flow cytometry were used to detect apoptotic rates of HepG2 cells; enzyme linked immunosorbent assay was used to detect the expression levels of Bax,bcl-2,and Caspase-3.Results:Compared with blank control group,the inhibitory rates of growth of HepG2 cells in middle and high doses of HCBF groups were increased (P <0.05 or P <0.01).The cells were shrank with irregular shape,and the nucleus was condensed and broken in HCBF group under inverted microscope.Compared with blank control group,the apoptotic rates of HepG2 cells in middle and high doses of HCBF groups were increased (P < 0.05 or P < 0.01).Compared with blank control group,the expression levels of bcl-2 protein in middle and high doses of HCBF groups were decreased (P <0.05 or P < 0.01),and the expression levels of Bax protein were significantly increased (P < 0.05 or P <0.01),and the expression level of Caspase-3 protein in high dose of HCBF group was significantly increased (P <0.05).Conclusion:The serum containing HCBF could induce the apoptosis of HepG2 cells,and its mechanism may be related to mitochondrial pathway.