异常黑胆质成熟剂总黄酮对人宫颈癌 HeLa 细胞增殖的影响

摘要

目的：探讨异常黑胆质成熟剂（ASMq）总黄酮对人宫颈癌 HeLa 细胞增殖的影响，为 ASMq 的利用和开发提供理论依据。方法采用常规细胞株及细胞培养、传代方法，对 HeLa 细胞进行培养、传代，取对数生长期细胞并进行药物干预，通过 MTT（检测细胞存活和生长的方法）探讨其抑制人宫颈癌 HeLa 细胞增殖的作用，倒置显微镜观察细胞形态学变化，荧光倒置显微镜观察细胞凋亡的动态情况。检测 ASMq 总黄酮作用下 HeLa 细胞总抗氧化能力（T-AOC）、谷胱甘肽（GSH）及总超氧化物歧化酶（T-SOD）的含量。结果MTT 实验结果表明，不同时间、不同浓度的 ASMq 总黄酮均具有抑制人宫颈癌 HeLa 细胞增殖的作用，其中浓度为400μg／mL、作用时间为72 h 时，作用较为明显。倒置显微镜细胞形态学观察可见，给药组中悬浮细胞增多，细胞质混浊，细胞胞体缩小、变圆、皱缩，核固缩碎裂，细胞折光性减弱，细胞内出现颗粒样物质，培养液中有较多的细胞碎片。荧光倒置显微镜观察可见，ASMq 总黄酮浓度为100μg／mL 时，随着时间的延长细胞发生的凋亡现象较为明显。当作用时间为24 h时，随着浓度的增高细胞发生的凋亡现象较为明显；ASMq 总黄酮作用于 HeLa 细胞24、48、72 h 后，不同剂量均增加 HeLa 细胞 T-AOC 及 T-SOD、GSH 含量，差异有统计学意义（P ＜0．05），其中高剂量组较为明显。结论ASMq 总黄酮对人宫颈癌 HeLa 细胞增殖具有抑制作用，能够增强人宫颈癌 HeLa 细胞的抗氧化能力。%Objective To observe the effect of total flavonoids of Abnormal Savda Munziq (ASMQ )on He-La human cervical carcinoma cell lines,so as to provide a theoretical basis for the utilization and develop-ment of ASMQ.Methods The HeLa cells were cultured using conventional method and the inhibition of human cervical cancer HeLa cell proliferation was investigated by the MTT method.The changes in the cell morphology was observed by inverted microscopy;fluorescence inverted microscope was used to ob-serve the changes of cell dynamics.The total antioxidant capacity in HeLa cells (T-AOC),glutathione (GSH) and total superoxide dismutase (T-SOD) content were detected by UV spectrophotometric method.Results The results of MTTN showed HeLa inhibited cell proliferation at different times with different concentration.There was the most obvious effect with the concentration of 400 ug/mL at the time of 72 h.Morphological features of cells showed obvious cell suspension in drug group increased,the cell cytoplasm was turbid,the cell body was narrow,rounded,shrinkage,nuclear condensation and fragmen-tation,and cell refractivity declined,granule was appeared in the cell body,and more cell debris was ob-served in the cell culture medium.Inverted fluorescence microscope showed that,when the concentration of ASMQ was 100 g/mL,with the extension of time,the apoptosis cells were occurred more obvious.When the reaction time was 24 h,as the concentration increased,the occurrence of cell apoptosis phenomenon is obvious;total flavonoids ASMQ effected on the cells at the time of 24,48,72 h,the content of T-AOC, T-SOD,and GSH in HeLa cells was increased,the difference was statistically significant (P <0.05),the high dosage group had the most obvious effect.Conclusion Total flavonoids of ASMQ can inhibit the pro-liferation and enhance the antioxidant ability of human cervix cancer HeLa cell.