基于p53/SLC7A11轴探讨补肾活血颗粒含药血清对MPP+诱导的PC12细胞铁死亡的影响
Effects of Bushen Huoxue Granules serum on ferroptosis in MPP+-induced PC12 cells based on the p53/SLC7A11 axis
谌盈帆 1栾振先 2齐小荣 1陈琦 2李绍旦 2杨明会2
作者信息
- 1. 解放军总医院第六医学中心中医医学部,北京 100048;解放军医学院,北京 100853
- 2. 解放军总医院第六医学中心中医医学部,北京 100048
- 折叠
摘要
目的:探讨补肾活血颗粒(BSHXG)血清对MPP+诱导的PC12细胞p53/SLC7A11轴及铁死亡的影响.方法:制备BSHXG含药血清和空白血清,体外培养PC12细胞,采用MPP+诱导建立帕金森病(PD)模型.设立空白组、MPP+组、BSHXG低剂量组(含5％含药血清)、BSHXG中剂量组(含10％含药血清)、BSHXG高剂量组(含20％含药血清)和铁抑素(Fer-1)组(含4 µmol/L Fer-1)6个组.采用Calcein AM/PI双染色法观察细胞存活率;CCK-8法检测细胞增殖活力;透射电镜检测细胞线粒体形态;FerroOrange亚铁离子(Fe2+)荧光探针检测Fe2+水平;RT-PCR和Western Blot分别检测p53、SLC7A11、PTGS2、GPX4、ACSL4 mRNA及蛋白表达水平.结果:与MPP+组比较,BSHXG含药血清和Fer-1可提高PC12细胞存活率和增殖活力;可明显改善细胞线粒体结构形态,降低细胞Fe2+水平;下调细胞p53、PTGS2、ACSL4 mRNA和蛋白的表达(P＜0.05,P＜0.01),上调SLC7A11、GPX4 mRNA和蛋白的表达(P＜0.05,P＜0.01).结论:BSHXG含药血清可通过调控p53/SLC7A11轴的表达,抑制铁死亡,从而减轻细胞损伤.
Abstract
Objective:To investigate the effects of Bushen Huoxue Granules(BSHXG)on p53/SLC7All axis and ferroptosis in PC12 cells induced by MPP+.Methods:BSHXG drug-containing serum and blank serum were prepared,and PC12 cells were cultured in vitro to establish a Parkinson's disease(PD)model using MPP+induction.Six groups were established:Blank group,MPP+group,BSHXG low-dose group(containing 5％BSHXG serum),BSHXG medium-dose group(containing 10％BSHXG serum),BSHXG high-dose group(containing 20％BSHXG serum)and ferrostatin-1(Fer-1)group(containing 4 μmol/L Fer-1).Cell survival ratio was observed by Calcein AM/PI double staining;cell proliferation viability was detected by CCK-8 assay;mitochondrial morphology of the cells was detected by transmission electron microscopy;intracellular Fe2+levels were detected using the FerroOrange fluorescent probe.RT-PCR and Western Blot were used to detect the expression of p53,SLC7A11,PTGS2,GPX4,and ACSL4 mRNA and protein,respectively.Results:Compared with the MPP+group,BSHXG drug-containing serum and Fer-1 increased the survival rate and proliferative viability of PC 12 cells;significantly improved the structural morphology of cellular mitochondria,decreased cellular Fe2+level;and down-regulated cellular p53,PTGS2,ACSL4 mRNA and protein expression(P＜0.05,P＜0.01),and up-regulated SLC7A11,GPX4 mRNA and protein expression(P＜0.05,P＜0.01).Conclusion:BSHXG-containing serum can inhibit ferroptosis by down-regulating the expression of p53/SLC7A11 axis,thus reducing cell damage.
关键词
补肾活血颗粒/帕金森病/铁死亡/含药血清/p53/SLC7A11轴Key words
Bushen Huoxue Granules/Parkinson disease/Ferroptosis/Drug-containing serum/p53/SLC7A11 axis引用本文复制引用
基金项目
国家自然科学基金重点项目(82130117)
出版年
2024
NETL
NSTL
万方数据