Abstract:

This study aims to investigate the mechanism of plumbagin (PLB) on alleviating sepsis-associated acute kidney injury (SA-AKI) via inhibiting JAK-STAT signaling pathway mediated ferroptosis based on network pharmacology and experimental validation. The target groups of PLB and SA-AKI were screened by network pharmacology and bioinformatics methods, and molecular docking was employed to validate the interactions between the core component and the core targets. The mouse SA-AKI model was induced by cecal ligation and puncture (CLP) to validate the related mechanisms. Renal tissue morphological changes were observed via HE staining. Serum levels of creatinine (CRE) and urea nitrogen (BUN), as well as the kidney tissue levels of malondialdehyde (MDA) and superoxide dismutase (SOD) were measured. Renal reactive oxygen species (ROS) levels were detected using DHE fluorescent probe. Serum levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were measured by ELISA Method. The protein expression levels of Janus kinase 2 (JAK2), phosphorylated JAK2 (p-JAK2), signal transducer and activator of transcription 3 (STAT3), phosphorylated STAT3 (p-STAT3), and cystine/glutamate antiporter xc^- system (xCT), glutathione peroxidase 4 (GPX4), ferroptosis suppressor protein 1 (FSP1) in kidney tissue were detected by Western blot. Network pharmacology analysis identified JAK2 and STAT3 as the core targets, molecular docking further verified that PLB could bind to both JAK2 and STAT3. When compared with CLP group, in the different doses of PLB groups, HE staining showed that the different doses of PLB ameliorated the pathological changes of kidney tissue. PLB treatment reduced the levels of renal function indicators (CRE, BUN), inflammatory factors (TNF-α, IL-1β), and oxidative stress levels (decreased MDA, increased SOD, decreased ROS). It also downregulated the protein expression of p-STAT3 and p-JAK2, while upregulating the expression of ferroptosis-related proteins xCT, GPX4, and FSP1. In conclusion, PLB may alleviate SA-AKI by inhibiting JAK-STAT signaling pathway mediated ferroptosis in mice model.

Key words:

network pharmacology, plumbagin; sepsis-induced acute kidney injury, ferroptosis, JAK-STAT signaling pathway