Effects of Bushen Huoxue Granules serum on ferroptosis in MPP+-induced PC12 cells based on the p53/SLC7A11 axis
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.
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