pf 543  (Pfizer Inc)

Journal: Frontiers in Immunology

Article Title: Liu Shen Wan regulates the SPHK1/S1P axis to ameliorate influenza-induced inflammation via integrated network pharmacology and lipidomics

doi: 10.3389/fimmu.2025.1764754

Figure Lengend Snippet: LSW inhibited sphingolipid metabolism in vitro . (A) The mRNA expression of ASMase in A549 cells during PR8 infection. (B) The mRNA expression of SPHK1 in A549 cells during PR8 infection. (C) The protein expression of SPHK1 in A549 cells during PR8 infection. (D) The relative expression of SPHK1 analyzed by Image J. (E) The protein expression of SPHK1 in A549 cells during PR8 infection; Scale bar = 50 μm. (F) The relative expression of SPHK1 analyzed by Image-Pro Plus 6.0. (G) The mRNA expression of SPHK1 in A549 cells stimulated by TNF-α. (H) The mRNA expression of MCP-1 in A549 cells stimulated by TNF-α. (I) The mRNA expression of CXCL10 in A549 cells stimulated by TNF-α. (J) The mRNA expression of IL6 in A549 cells stimulated by TNF-α. The data were shown as mean ± SD and analyzed by one-way ANOVA Bonferroni or Dunnett’s multiple comparisons tests (n=3). *, p < 0.05 or ***, p < 0.001. vs . PR8 group.

Article Snippet: PF-543 hydrochloride (lot: S7177), a SPHK1 inhibitor, was purchased from Selleck.

Techniques: In Vitro, Expressing, Infection

Journal: Frontiers in Immunology

Article Title: Liu Shen Wan regulates the SPHK1/S1P axis to ameliorate influenza-induced inflammation via integrated network pharmacology and lipidomics

doi: 10.3389/fimmu.2025.1764754

Figure Lengend Snippet: LSW inhibited SPHK1/S1P axis in vivo during PR8 infection. (A) The protein expression of SPHK1 in lungs during PR8 infection. (B) The protein expression and localization of SPHK1 in lungs during PR8 infection; Scale bar = 100 μm. (C) The relative expression of SPHK1 analyzed by Image-Pro Plus 6.0. (D) The production of S1P in lungs during PR8 infection. (E) The production of S1P in sera during PR8 infection. The data were shown as mean ± SD and analyzed by one-way ANOVA Bonferroni or Dunnett’s multiple comparisons tests (n=3). *, p < 0.05; **, p < 0.01 or ***, p < 0.001. vs . PR8 group.

Article Snippet: PF-543 hydrochloride (lot: S7177), a SPHK1 inhibitor, was purchased from Selleck.

Techniques: In Vivo, Infection, Expressing

Journal: Frontiers in Immunology

Article Title: Liu Shen Wan regulates the SPHK1/S1P axis to ameliorate influenza-induced inflammation via integrated network pharmacology and lipidomics

doi: 10.3389/fimmu.2025.1764754

Figure Lengend Snippet: Inhibition of SPHK1 reduced the expression of cytokines and chemokines during PR8 infection. (A) The mRNA expression of MCP-1 in A549 cells during PR8 infection. (B) The mRNA expression of CXCL10 in A549 cells during PR8 infection. (C) The mRNA expression of TNF-α in A549 cells during PR8 infection. (D) The mRNA expression of IL6 in A549 cells during PR8 infection. (E) The protein expression of SPHK1 in A549 cells during PR8 infection; Scale bar = 50 μm. (F) The relative expression of SPHK1 analyzed by Image-Pro Plus 6.0. The data were shown as mean ± SD and analyzed by one-way ANOVA Bonferroni or Dunnett’s multiple comparisons tests (n=3). ***, p < 0.001. vs . PR8 group.

Article Snippet: PF-543 hydrochloride (lot: S7177), a SPHK1 inhibitor, was purchased from Selleck.

Techniques: Inhibition, Expressing, Infection

Journal: Frontiers in Immunology

Article Title: Liu Shen Wan regulates the SPHK1/S1P axis to ameliorate influenza-induced inflammation via integrated network pharmacology and lipidomics

doi: 10.3389/fimmu.2025.1764754

Figure Lengend Snippet: LSW inhibited the expression of SPHK1, cytokines and chemokines induced by SPHK1 overexpression. (A) The mRNA expression of SPHK1 induced by SPHK1 overexpression in A549 cells. (B) The mRNA expression of CXCL10 induced by SPHK1 overexpression in A549 cells. (C) The mRNA expression of MCP-1 induced by SPHK1 overexpression in A549 cells. (D) The mRNA expression of IL6 induced by SPHK1 overexpression in A549 cells. (E) H&E staining of lungs after delivery of rAAV; Scale bar = 500 μm. (F) The expression of SPHK1 in lungs after delivery of rAAV. (G) The expression of TNF-α in lungs after delivery of rAAV. (H) The expression of IFN-γ in lungs after delivery of rAAV. The data were shown as mean ± SD and analyzed by one-way ANOVA Bonferroni or Dunnett’s multiple comparisons tests (n=3). **, p < 0.01 or ***, p < 0.001. vs . PR8 group.

Article Snippet: PF-543 hydrochloride (lot: S7177), a SPHK1 inhibitor, was purchased from Selleck.

Techniques: Expressing, Over Expression, Staining

Journal: Frontiers in Immunology

Article Title: Liu Shen Wan regulates the SPHK1/S1P axis to ameliorate influenza-induced inflammation via integrated network pharmacology and lipidomics

doi: 10.3389/fimmu.2025.1764754

Figure Lengend Snippet: The interaction of the key compound with SPHK1. (A) The interaction of bufalin (golden) with SPHK1 (gray); (B) The interaction of bufotalin (golden) with SPHK1 (gray); (C) The interaction of decamine (golden) with SPHK1 (gray); (D) The interaction of ursolic acid (golden) with SPHK1 (gray).

Article Snippet: PF-543 hydrochloride (lot: S7177), a SPHK1 inhibitor, was purchased from Selleck.

Techniques:

Journal: Frontiers in Immunology

Article Title: Liu Shen Wan regulates the SPHK1/S1P axis to ameliorate influenza-induced inflammation via integrated network pharmacology and lipidomics

doi: 10.3389/fimmu.2025.1764754

Figure Lengend Snippet: Schematic representation of the mechanisms of LSW on influenza virus infection. The sphingolipid signaling pathway was activated during influenza virus infection, accompanied with the release of cytokines and chemokines. LSW alleviated virus-induced overactivated inflammatory response by inhibiting SPHK1/S1P axis.

Article Snippet: PF-543 hydrochloride (lot: S7177), a SPHK1 inhibitor, was purchased from Selleck.

Techniques: Virus, Infection

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, HK-2 cells were treated with 2-DG (MCE, HY- D13966 , 5 mM), oxamate (MCE, HY-W013032A, 5 mM), Fer-1 (Selleck, S7243, 10 μM), 3-MA (MCE, HY-19312, 5 mM), Z-VAD-FMK (MCE, HY-16658B, 20 μM), AC-YVAD-CMK (MCE, HY-16990, 40 μM), Nigericin (MCE, HY-127019, 2 μg/mL), PF-543 (MCE, HY-15425, 100 nM), C646 (MCE, HY-13823, 10 μM), Sanguinarine (MCE, HY-N0052, 2 μM), Quercetin (MCE, HY-18085, 4 μM), NAD + (MCE, HY-B0445, 200 μM), EX-527 (Selleck, S1541, 10 μM), and Resveratrol (Selleck, S1396, 10 μM).

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, HK-2 cells were treated with 2-DG (MCE, HY- D13966 , 5 mM), oxamate (MCE, HY-W013032A, 5 mM), Fer-1 (Selleck, S7243, 10 μM), 3-MA (MCE, HY-19312, 5 mM), Z-VAD-FMK (MCE, HY-16658B, 20 μM), AC-YVAD-CMK (MCE, HY-16990, 40 μM), Nigericin (MCE, HY-127019, 2 μg/mL), PF-543 (MCE, HY-15425, 100 nM), C646 (MCE, HY-13823, 10 μM), Sanguinarine (MCE, HY-N0052, 2 μM), Quercetin (MCE, HY-18085, 4 μM), NAD + (MCE, HY-B0445, 200 μM), EX-527 (Selleck, S1541, 10 μM), and Resveratrol (Selleck, S1396, 10 μM).

Techniques: Staining