Tubular injury and oxidative stress are involved in the pathogenesis of diabetic kidney disease (DKD). Astragaloside IV (ASIV) is a natural antioxidant. The effects and underlying molecular mechanisms of ASIV on DKD have not been elucidated. The db/db mice and high-glucose-stimulated HK2 cells were used to evaluate the beneficial effects of ASIV in vivo and in vitro. Succinylated proteomics was used to identify novel mechanisms of ASIV against DKD and experimentally further validated. ASIV alleviated renal dysfunction and proteinuria, downregulated fasting blood glucose, and upregulated insulin sensitivity in db/db mice. Meanwhile, ASIV alleviated tubular injury, oxidative stress, and mitochondrial dysfunction in vivo and in vitro. Mechanistically, ASIV reversed downregulated 17beta-hydroxysteroid dehydrogenase type 10 (HSD17B10) lysine succinylation by restoring carnitine palmitoyl-transferase1alpha (Cpt1a or CPT1A) activity in vivo and in vitro. Molecular docking and cell thermal shift assay revealed that ASIV may bind to CPT1A. Molecular dynamics simulations demonstrated K99 succinylation of HSD17B10 maintained mitochondrial RNA ribonuclease P (RNase P) stability. The K99R mutation of HSD17B10 induced oxidative stress and disrupted its binding to CPT1A or mitochondrial ribonuclease P protein 1 (MRPP1). Importantly, ASIV restored the interaction between HSD17B10 and MRPP1 in vivo and in vitro. We also demonstrated that ASIV reversed high-glucose-induced impaired RNase P activity in HK2 cells, which was suppressed upon K99R mutation of HSD17B10. These findings suggest that ASIV ameliorates oxidative stress-associated proximal tubular injury by upregulating CPT1A-mediated K99 succinylation of HSD17B10 to maintain RNase P activity.
Triphenyltin chloride (TPTC), a member of the organotin family, has been widely restricted due to its endocrine-disrupting effects. Nevertheless, TPTC continues to be detected in aquatic environments and poses potential health risks owing to its persistence and bioaccumulation. Although previous studies have shown that triphenyltin compounds can disrupt lipid homeostasis through nuclear receptor pathways involving retinoid X receptor (RXR) and peroxisome proliferator-activated receptors (PPARs), whether TPTC exerts hepatocytotoxicity through direct metabolic targets remains unclear. This study employed a metabolomics-driven target discovery strategy to analyze intracellular metabolite extracts from HepG2 cells exposed to TPTC for 24 h, aiming to investigate the underlying molecular mechanisms of TPTC-induced metabolic toxicity. Untargeted global metabolomics analysis revealed that TPTC exposure caused significant disturbances in hepatic lipid metabolism, characterized by the accumulation of medium- and long-chain fatty acids and depletion of key metabolites associated with fatty acid β-oxidation. Pathway enrichment analysis and protein–protein interaction network analysis further indicated that two mitochondrial dehydrogenases involved in fatty acid catabolism─hydroxysteroid 17β-dehydrogenase 10 (HSD17B10) and hydroxyacyl-CoA dehydrogenase (HADH)─are potential metabolic targets responsible for the observed alterations. Molecular docking, surface plasmon resonance, and cellular thermal shift assays confirmed direct interactions between TPTC and both HSD17B10 and HADH, with binding affinities at the micromolar level. Functional validation demonstrated that TPTC disrupts mitochondrial respiratory function and redox balance by interfering with HADH- and HSD17B10-dependent generation of acetyl-CoA and NADH, leading to mitochondrial dysfunction and oxidative stress. In summary, this study reveals a previously underappreciated mechanism of TPTC toxicity involving mitochondrial metabolism, extending the current understanding of its toxicological pathways beyond nuclear receptor-mediated signaling to include direct disruption of mitochondrial energy and redox metabolism.
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