二氢青蒿素对衰老细胞的作用及机制

目的 探究二氢青蒿素对衰老细胞的作用及其机制。方法 利用双氧水诱导小鼠成纤维细胞(NIH3T3)构建应激早衰模型(SIPS),通过衰老特异性标记物SA-β-半乳糖苷酶染色、蛋白质印迹技术进行SIPS鉴定；细胞实验分为对照组(磷酸盐缓冲液+二甲基亚砜)、模型组(双氧水+二甲基亚砜)、药物组(双氧水+二氢青蒿素),采用Cell Counting Kit-8(CCK-8)法、乳酸脱氢酶法、SA-β-半乳糖苷酶染色、细胞内铁含量测定、蛋白质印迹技术、荧光探针染色及流式细胞术检测二氢青蒿素对衰老细胞的影响。结果 CCK-8结果显示，二氢青蒿素作用模型组24 h的半数抑制浓度(IC50)为(8.26±0.66)μmol/L;与对照组相比，模型组存活率降低，且呈剂量依耐性；蛋白质印迹结果显示，与模型组相比，药物组铁蛋白重链(FTH)及谷胱甘肽过氧化酶4(GPX4)蛋白表达水平降低(P<0.05),细胞内铁、活性氧(ROS)含量增加(P<0.05)。结论 二氢青蒿素可特异性诱导衰老细胞死亡，其机制与铁死亡有关。

The effect and mechanism of dihydroartemisinin on aging cells

Objective To study the regulation of dihydroartemisinin on senescent cells and its possible antr-aging mechanismMethods A stressinduccd premature senescence(SIPS)model of mouse fibroblast NIH3T3 cells was established by hydrogenperoxide.And the SIPS was detected by senescing-specific marker SA-P-galactosidase staining,and western blotting.Therewere 3groups in the cll experiment:control group(phosphate buffer solution +dimethyl sulfoxide),model group(hydrogen peroxide+dimethyl sulfoxide),druggroup(hydrogen peroxide +dihydroartemisinin).The effect of dihydroartemisinin on senescent clls wasdetected by CCK-8,lactate dehydrogenaseassay,SA-P-galactosidase staining,intracellular iron content determination,Westernblotting,fluorescent probe staining and flow cytometry.Results The results of CCK-8 showed that the half maximal inhibitoryconcentration (IC50)of the dihydroartemisinin treated group was(8.26±0.66)μmol/L at 24 h.Compared with the control group,the cell survival rate in the model group was decreased and showed dose-dependent tolerance.Westen bloting results showed thatcompared with themodel group,theprotein expression levels offerritin heavy chain(FTH)andglutathione peroxidase 4(GPX4)inthe drug group were decreased (P<0.05),and the contents of intracellular iron and reactive oxygen species(ROS)wereincreased(P<0.05).Conclusion Dihydroartemisinin can specificlly induce senescent cell death,and its mechanism is related to ferroptosis