重楼皂苷Ⅱ诱导黑色素瘤细胞B16F10铁死亡的作用研究  

Ferroptosis in melanoma cells B16F10 induced by polyphyllin Ⅱ

作　　者：陈馨 刘霞霞 王文君 王萌[1,2] 程卉 CHEN Xin;LIU Xia-xia;WANG Wen-jun;WANG Meng;CHENG Hui(School of Pharmacy,Anhui University of Chinese Medicine,Hefei 230012;Research and Technology Center,Anhui University of Chinese Medicine,Hefei 230038)

机构地区：[1]安徽中医药大学药学院,合肥230012 [2]安徽中医药大学科研技术中心,合肥230038

出　　处：《中南药学》2023年第2期279-284,共6页Central South Pharmacy

基　　金：国家自然科学基金资助项目(No.81903859)。

摘　　要：目的 探讨重楼皂苷Ⅱ对黑色素瘤细胞B16F10的抑制作用及可能的作用机制。方法四甲基偶氮唑蓝(MTT)法检测不同浓度的重楼皂苷Ⅱ对黑色素瘤细胞B16F10的抑制作用;透射电镜(TEM)观察细胞线粒体结构的变化;流式细胞仪检测活性氧(ROS)的变化;试剂盒检测铁死亡相关指标丙二醛(MDA)的含量、超氧化物歧化酶(SOD)的含量、铁(Fe)的含量及谷胱甘肽(GSH)的水平;Western blot法检测铁死亡相关蛋白的表达变化。结果 与空白组相比,重楼皂苷Ⅱ处理后B16F10细胞的活性显著降低,具有剂量和时间依赖性。TEM可以观察到重楼皂苷Ⅱ处理后B16F10细胞中线粒体结构发生皱缩变小以及膜密度增加的变化。重楼皂苷Ⅱ作用后ROS的水平、MDA和Fe的含量提高,SOD的含量和GSH的水平降低。除此之外,重楼皂苷Ⅱ可以抑制黑色素瘤细胞B16F10中GPX4、FTH-1、SLC7A11蛋白的表达水平而促进P53、ACSL4蛋白的表达水平。结论 重楼皂苷Ⅱ可以抑制黑色素瘤细胞B16F10增殖,其可能与诱导细胞发生铁死亡有关。Objective To determine the inhibitory effect of polyphyllinⅡ(PPⅡ) on melanoma B16F10 cells and possible mechanism. Methods MTT assay was used to detect the inhibitory effect of different concentrations of polyphyllinⅡ on melanoma cells B16F10. Transmission electron microscopy(TEM) was used to observe the changes of cell mitochondrial structure. Flow cytometry was used to detect the changes of reactive oxygen species(ROS). The ferroptosis-related indicators, including malondialdehyde(MDA) content, superoxide dismutase(SOD) content, iron(Fe) level and glutathione(GSH) level were detected by kits. Western blot was used to detect the changes of ferroptosis-related protein expression. Results Compared with the control group, the viability of B16F10 cells was significantly reduced after the polyphyllinⅡ treatment in a dose-and time-dependent manner. Under the transmission electron microscope, it can be observed that the mitochondrial structure shrank and the cell membrane density increased in B16F10 cells after the polyphyllinⅡtreatment. The ROS level and the content of MDA and Fe were increased, while the content of SOD and GSH was decreased after the polyphyllinⅡtreatment. PolyphyllinⅡinhibited the expression levels of GPX4, FTH-1 and SLC7A11 proteins and promoted the expression level of P53 and ACSL4 protein in melanoma cells B16F10. Conclusion PolyphyllinⅡ can inhibit the proliferation of melanoma cells B16F10 in mice, which may be related to the induction of ferroptosis.

关 键 词：黑色素瘤 重楼皂苷Ⅱ 铁死亡 

分 类 号：R285[医药卫生—中药学]