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: SPHK1 drives lactate-enhanced pyroptosis of RTEC in SA-AKI via activating NLRP3 inflammasome. ( A) Venn diagram of the intersection of five gene sets identified seven candidate genes linking lactate, pyroptosis, and SA-AKI. (B) Heatmap of candidate genes expression across datasets. (C) Protein levels of SPHK1 in HK-2LPS cells treated with lactate/2-DG/oxamate (n = 5). (D) Protein levels of GSDMD, Caspase-1, Cleaved Caspase-1, NLRP3, and SPHK1 and (E) ELISA analysis of IL-18 and IL-1β levels in HK-2LPS cells transfected with siNC or siSPHK1 under lactate challenge or not (n = 5). (F) Protein levels of GSDMD, Caspase-1, Cleaved Caspase-1, and NLRP3 and (G) ELISA analysis of IL-18 and IL-1β levels in HK-2LPS cells treated with lactate or lactate + PF-543 (n = 5). (H and I) mRNA and protein levels of SPHK1, and (J) representative H&E staining and IHC images of SPHK1 (scale bar = 50 μm) in kidney tissue of CLP mice treated with lactate/2-DG/oxamate (n = 5). (K) Protein levels of GSDMD, Caspase-1, Cleaved Caspase-1, NLRP3, and SPHK1 and (L) ELISA analysis of IL-18 and IL-1β levels in kidney tissue from WT or SPHK1-/- CLP mice under lactate challenge or not (n = 5). (M) Protein levels of GSDMD, Caspase-1, Cleaved Caspase-1, and NLRP3 and (N) ELISA analysis of IL-18 and IL-1β levels in CLP mice treated with lactate or lactate + PF-543 (n = 5). (O) Representative H&E staining and IHC images of KIM-1 and SPHK1 (scale bar = 50 μm) in kidney tissue of CLP mice. 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: Expressing, Enzyme-linked Immunosorbent Assay, Transfection, Staining

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: PF-543 and NAD+ synergistically alleviate progression of SA-AKI. (A) Protein levels of KIM-1, (B) cell viability, (C) representative morphological features of pyroptosis (bright-field, scale bar = 100 μm) and (D) LDH release in lactated-treated HK-2LPS cells pretreated with PF-543, NAD+, or PF-543 + NAD+ (n = 5). (E) Protein levels of KIM-1, (F) Scr, (G) BUN, (H) representative H&E staining and IHC images of KIM-1 in kidney tissue of CLP mice subjected to lactate pretreated with PF-543, NAD+, or PF-543 + NAD+ (n = 5). (E) Survival curves of CLP mice subjected to lactate pretreated with PF-543, NAD+, or PF-543 + NAD+ (n = 12). 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: Staining