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

作　　者：谌盈帆 栾振先 齐小荣 陈琦[1] 李绍旦[1] 杨明会[1] CHEN Yingfan;LUAN Zhenxian;QI Xiaorong;CHEN Qi;LI Shaodan;YANG Minghui(Department of Traditional Chinese Medicine,Sixth Medical Center of Chinese PLA General Hospital,Beijing 100048,China;Medical School of Chinese PLA,Beijing 100853,China)

机构地区：[1]解放军总医院第六医学中心中医医学部,北京100048 [2]解放军医学院,北京100853

出　　处：《中华中医药杂志》2024年第4期1735-1741,共7页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金重点项目(No.82130117)。

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4μmol/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(Fe^(2+))荧光探针检测Fe^(2+)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Fe^(2+)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。Objective:To investigate the effects of Bushen Huoxue Granules(BSHXG)on p53/SLC7A11 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 4μmol/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 Fe^(2+)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 drugcontaining serum and Fer-1 increased the survival rate and proliferative viability of PC12 cells;significantly improved the structural morphology of cellular mitochondria,decreased cellular Fe^(2+)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 

分 类 号：R285[医药卫生—中药学]