Journal: Theranostics

Article Title: H3K18 lactylation-mediated SPHK1-SIRT1 feedback loop accelerates pyroptosis of tubular epithelial cells in sepsis-associated acute kidney injury

doi: 10.7150/thno.122991

Figure Lengend Snippet: NAD+ emerges as a therapeutic candidate targeting SPHK1-driven kidney injury via activating SIRT1-mediated delactylation. (A) Schematic of high-throughput drug screening workflow in lactate-treated HK-2LPS cells. (B) Venn diagram identified 22 compounds that coordinately downregulate SPHK1 and KIM-1. (C) Top 3 candidates (NAD+, quercetin, and sanguinarine) ranked by efficacy. (D and E) mRNA and protein levels of KIM-1 and SPHK1 in lactate-treated HK-2LPS cells pretreated with sanguinarine/quercetin/NAD+ (n = 5). (F and G) mRNA and protein levels of KIM-1 and SPHK1 in kidney tissue of CLP mice subjected to lactate pretreated with sanguinarine/quercetin/NAD+ (n = 5). (H) Protein levels of pan-Kla in lactate-treated HK-2 cells pretreated with NAD+, NAD+ + EX-527, or resveratrol (n = 5). (I) mRNA levels of SPHK1 and (J) protein levels of SPHK1 and H3K18la in lactate-treated HK-2 cells pretreated with NAD+, NAD+ + EX-527, or resveratrol (n = 5). Data are mean ± SEM. *p < 0.05, **p < 0.01, and ***p < 0.001; ns, not significant.

Article Snippet: In separate experiments, mice were treated with oxamate (MCE, HY-W013032A, 0.5 g/kg), 2-DG (MCE, HY- D13966 , 0.5 g/kg), AC-YVAD-CMK (MCE, HY-16990, 10 mg/kg), Nigericin (MCE, HY-127019, 1 mg/kg), PF-543 (MCE, HY-15425, 10 mg/kg), C646 (MCE, HY-13823, 10 mg/kg), Sanguinarine (MCE, HY-N0052, 10 mg/kg), Quercetin (MCE, HY-18085, 50 mg/kg), NAD + (MCE, HY-B0445, 100 mg/kg), EX-527 (Selleck, S1541, 10 mg/kg), and Resveratrol (Selleck, S1396, 50 mg/kg).

Techniques: High Throughput Screening Assay, Drug discovery