Rong Lin ^* Xiaoying Weng ^* Liang Lin ^* Xuyang Hu ^* Zhiyan Liu ^* Jing Zheng ^* Fenfang Shen ^* Rui Li ^*

• Shengli Clinical Medical College, Fujian Medical University, Fuzhou, China

1) Background: The involvement of mitochondrial and programmed cell death (mtPCD)-related genes in the pathogenesis of pre-eclampsia (PE) remains inadequately characterized. (2) Methods:This study explores the role of mtPCD genes in PE through bioinformatics and experimental approaches. Differentially expressed mtPCD genes (DE-mtPCD) were identified as potential biomarkers from the GSE10588 and GSE98224 datasets and subsequently validated. Hub genes were determined using SVM, LASSO, and Boruta based on consistent expression profiles. Their performance was assessed through nomogram and artificial neural network (ANN) models.Biomarkers were subjected to localization, functional annotation, regulatory network analysis, and drug prediction. Clinical validation was conducted via RT-qPCR, immunofluorescence, and Western Blot.blot. (3) Results: Four genes (SLC25A5, ACSF2, MFF, and PMAIP1) were identified as biomarkers distinguishing PE from normal controls. Functional analysis indicated their involvement in various biological pathways. Immune analysis revealed associations between biomarkers and immune cell activity. A regulatory network was informed by biomarker expression and database predictions, in which KCNQ1OT1 modulates ACSF2 expression via hsa-miR-200b-3p. Drug predictions, including clodronic acid, were also proposed. Immunofluorescence, RT-qPCR, and Western blot confirmed reduced expression of SLC25A5, MFF, and PMAIP1 in PE, while ACSF2 was significantly upregulated. (4) Conclusion: These four mtPCD-related biomarkers may play a pivotal role in PE pathogenesis, offering new perspectives on the disease's diagnostic and mechanistic pathways.