impdh2  (R&D Systems)

Journal: Redox Biology

Article Title: Alpha-ketoglutarate accelerates granulocyte-monocyte progenitor differentiation and atherosclerotic plaque inflammation via oxoglutarate receptor 1

doi: 10.1016/j.redox.2026.104134

Figure Lengend Snippet: PNP promoted GMP proliferation and enhanced myeloid cell production via inosine-5′-monophosphate dehydrogenase 2 (IMPDH2). Violin plot presenting PNP expression in GMPs and subpopulations by scRNA-seq analysis. ( A–F ) BMCs were exposed to PNP (0–1 U/mL) for 24 and 72 h. FACS analysis of the frequency of HSPCs, GMPs, and Pro-NEU. ( G–I ) After 3 days of PNP treatment, the proportion of neutrophils and monocytes in BMCs was quantified by FACS. ( J–Q ) Lineage −/low cells were treated with 0 or 0.1 U/mL PNP for 24 h. Ribose-5-phosphate and IMPDH2 expression was measured by ELISA. BMCs were treated with PNP in the absence or presence of IMPDH2 inhibitor for 24 h. EDU + GMPs were analyzed by FACS. In parallel, the frequency of HSPCs, GMP, Pro-NEU, and cMOP was analyzed by FACS. ( R–S ) Lineage −/low cells were treated with 0 or 0.1 U/mL PNP for 72 h. Neutrophil and monocyte production were evaluated by FACS. BMC: bone marrow cell; ELISA: enzyme-linked immunosorbent assay; FACS: fluorescence-activated cell sorting; GMP: granulocyte-monocyte progenitor; HSPC: hematopoietic stem/progenitor cell; Pro-NEU, Pro-neutrophils; cMOP, common monocyte progenitors; IMPDH2: inosine-5′-monophosphate dehydrogenase 2; PNP: purine nucleoside phosphorylase; scRNA-seq: single-cell RNA sequencing. Two experimental groups were compared using the unpaired two-tailed Student's t-test. In the experiments involving ≥3 groups, one-way analysis of variance followed by Dunnett's test was used.

Article Snippet: Protein was extracted from PNP-treated lineage −/low cells to determine the expression of ribose-5-phosphate, IMPDH2, NAD, SOD, and ATP by ELISA, following the manufacturer's instructions (mouse IMPDH2 ELISA kit [#ml601123], mouse NAD ELISA kit [# JK691233 ], Enzyme-linked Biotechnology, China; mouse SOD ELISA kit [#CSB- E08556 m], Huamei Bioengineering, China; ATP Assay Kit [#HY-K0314], MedChemExpress, US).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Fluorescence, FACS, Single Cell, RNA Sequencing, Two Tailed Test

Journal: Redox Biology

Article Title: Alpha-ketoglutarate accelerates granulocyte-monocyte progenitor differentiation and atherosclerotic plaque inflammation via oxoglutarate receptor 1

doi: 10.1016/j.redox.2026.104134

Figure Lengend Snippet: Correlation of LDL-c and TCA metabolites in human plasma samples. (A-B) The fasting plasma levels of LDL-c and isocitrate were determined in healthy human subjects (n = 40) and their correlation was analyzed by Spearman's correlation analysis. ATP: adenosine triphosphate; ELISA: enzyme-linked immunosorbent assay; GPX: glutathione peroxidase; LDL-c: low-density lipoprotein cholesterol; NAD: nicotinamide adenine dinucleotide; PNP: purine nucleoside phosphorylase; ROS: reactive oxygen species; SOD: superoxide dismutase. (C) A proposed model of HFD-induced skewed myelopoiesis and atherosclerosis via α-ketoglutarate. In mice fed a HFD, isocitrate was elevated and LDL-c increased isocitrate dehydrogenase expression, leading to enhanced α-ketoglutarate production. Acting through OXGR1, α-ketoglutarate upregulated PNP mRNA and protein expression. PNP catabolized guanosine/deoxyguanosine and inosine/deoxyinosine, generating ribose-1-phosphate. By interconversion, ribose-5-phosphate production was increased and initiated de novo purine biosynthesis, resulting in IMPDH2 production and GMP proliferation. PNP metabolized nicotinamide riboside to Nam, decreasing NMN and NAD production. PNP increased NAD kinase expression to further accelerate NAD consumption. Furthermore, PNP increased inflammatory protein expression such as NF-κB. All of which reduced NAD production promoted ROS production and inflammation to facilitate GMP differentiation. Ultimately, the pronounced GMP proliferation and differentiation led to inflammatory myeloid cell production and atherosclerotic progression. GMP: granulocyte-monocyte progenitor; HFD: high-fat diet; IMPDH2: inosine-5′-monophosphate dehydrogenase 2; LDL-c: low-density lipoprotein cholesterol; NAD: nicotinamide adenine dinucleotide; OXGR1: oxoglutarate receptor 1; PNP: purine nucleoside phosphorylase; ROS: reactive oxygen species.

Article Snippet: Protein was extracted from PNP-treated lineage −/low cells to determine the expression of ribose-5-phosphate, IMPDH2, NAD, SOD, and ATP by ELISA, following the manufacturer's instructions (mouse IMPDH2 ELISA kit [#ml601123], mouse NAD ELISA kit [# JK691233 ], Enzyme-linked Biotechnology, China; mouse SOD ELISA kit [#CSB- E08556 m], Huamei Bioengineering, China; ATP Assay Kit [#HY-K0314], MedChemExpress, US).

Techniques: Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Expressing