Journal: Frontiers in Pharmacology

Article Title: Indoleamine 2,3-dioxygenase-regulated macrophages metabolic reprogramming rescues tacrolimus-induced nephrotoxicity

doi: 10.3389/fphar.2026.1784153

Figure Lengend Snippet: TAC Specifically Inhibits IDO1 Expression to Induce Kidney Injury (A) Relative abundances of Trp metabolites in mouse plasma (n = 5). (B) Relative abundances of Trp metabolites in the renal cortex (n = 5). (C) Quantification of Trp, KYN, the KYN/Trp ratio, and NAD + levels in renal tissues (n = 5). (D) Western blot analysis of IDO1 protein expression in the renal cortex, with β-tubulin as a loading control. (E) Densitometric quantification of IDO1 protein bands (n = 3). (F) qPCR analysis of IDO1 mRNA expression in the renal cortex (n = 3). (G) Molecular docking model of TAC binding to IDO1. (H) Schematic illustration showing that IDO1 inhibition disrupts the Trp–KYN pathway, affecting downstream metabolites including 3-HK, KYNA, 3-HAA, and QA. Data are presented as mean ± SD. Statistical analyses were performed using one-way ANOVA followed by Tukey’s multiple comparisons test. *P < 0.05, **P < 0.01, ***P < 0.001. Abbreviations: TRP, tryptophan; KYN, kynurenine; KYNA, kynurenic acid; QA, quinolinic acid; XA, xanthurenic acid; PIC, picolinic acid; 3-HK, 3-hydroxykynurenine; 3-HAA, 3-hydroxyanthranilic acid; IPYA, indole-3-pyruvic acid; IPA, indole-3-propionic acid; ILA, indolelactic acid; IALD, indoleacetaldehyde; IAA, indoleacetic acid; NAD + , nicotinamide adenine dinucleotide.

Article Snippet: Primary antibodies used in this study included: IDO1 (Proteintech, Cat# 13268-1-AP, 1:1000), CPT1 (ABclonal, Cat# A5307, 1:2000), CPT2 (Proteintech, Cat# 26555-1-AP, 1:8000), CACT (ABclonal, Cat# A13956, 1:400), β-tubulin (Proteintech, Cat# 80713-1-RR, 1:10,000), HRP-conjugated Goat Anti-Rabbit IgG (H + L) (ABclonal, Cat# SA00001-2, 1:10,000).

Techniques: Expressing, Clinical Proteomics, Western Blot, Control, Binding Assay, Inhibition

Journal: Frontiers in Pharmacology

Article Title: Indoleamine 2,3-dioxygenase-regulated macrophages metabolic reprogramming rescues tacrolimus-induced nephrotoxicity

doi: 10.3389/fphar.2026.1784153

Figure Lengend Snippet: Indicators of renal function and energy metabolism upon IDO1 inhibition. (A) Biochemical indicators of kidney injury in mice, including urinary protein, serum creatinine, blood urea nitrogen, uric acid, and hemoglobin (n = 8). (B) Representative H&E staining of kidney sections and corresponding tubular injury scores (n = 5). Arrows highlight tubular injury features. Scale bar = 50 μm. (C,D) Targeted analysis of TCA cycle metabolites (C) and glycolytic metabolites (D) in HEK293T cells. (E,F) Targeted analysis of TCA cycle metabolites (E) and glycolytic metabolites (F) in iBMDMs. (G) Targeted measurement of fatty acid–related metabolites in iBMDMs. (H) OCR trace of iBMDMs measured using glucose as the substrate during a Seahorse mitochondrial stress test (n = 6). (I) OCR trace of iBMDMs measured using palmitic acid as the substrate during a Seahorse mitochondrial stress test (n = 6). (J) Quantification of basal and maximal OCR in iBMDMs using glucose or palmitic acid as substrates. (K) ECAR trace of iBMDMs during glycolysis stress testing (n = 6). (L) Quantification of basal and maximal ECAR in iBMDMs. Data are presented as mean ± SD. Statistical analyses were performed using one-way ANOVA followed by Tukey’s multiple comparisons test. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Primary antibodies used in this study included: IDO1 (Proteintech, Cat# 13268-1-AP, 1:1000), CPT1 (ABclonal, Cat# A5307, 1:2000), CPT2 (Proteintech, Cat# 26555-1-AP, 1:8000), CACT (ABclonal, Cat# A13956, 1:400), β-tubulin (Proteintech, Cat# 80713-1-RR, 1:10,000), HRP-conjugated Goat Anti-Rabbit IgG (H + L) (ABclonal, Cat# SA00001-2, 1:10,000).

Techniques: Inhibition, Staining

Journal: Frontiers in Pharmacology

Article Title: Indoleamine 2,3-dioxygenase-regulated macrophages metabolic reprogramming rescues tacrolimus-induced nephrotoxicity

doi: 10.3389/fphar.2026.1784153

Figure Lengend Snippet: Indicators of macrophage polarization and proinflammatory cytokine expression. (A) Double immunofluorescence staining of CD68 (red) and ARG1 (green) in the renal cortex. (B) Double immunofluorescence staining of CD68 (red) and CD86 (green) in the renal cortex. (C) Quantification of CD86 + /CD68 + (left) and ARG1 + /CD68 + (right) double-positive cells (n = 3). (D) mRNA expression levels of M1-and M2-associated genes (CXCL9, IL-1β, IL-6, iNOS, TNF-α, ARG1, IL-10) in the renal cortex (n = 3). (E) mRNA expression levels of M1-and M2-associated genes in iBMDMs (n = 3). (F) mRNA expression levels of IDO1, ARG1, and iNOS in primary renal macrophages under indicated treatments (n = 3). Data are presented as mean ± SD. Statistical analyses were performed using one-way ANOVA followed by Tukey’s multiple comparisons test. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Primary antibodies used in this study included: IDO1 (Proteintech, Cat# 13268-1-AP, 1:1000), CPT1 (ABclonal, Cat# A5307, 1:2000), CPT2 (Proteintech, Cat# 26555-1-AP, 1:8000), CACT (ABclonal, Cat# A13956, 1:400), β-tubulin (Proteintech, Cat# 80713-1-RR, 1:10,000), HRP-conjugated Goat Anti-Rabbit IgG (H + L) (ABclonal, Cat# SA00001-2, 1:10,000).

Techniques: Expressing, Double Immunofluorescence Staining