Peptidoglycan-Mediated Bone Marrow Autonomic Neuropathy Impairs Hematopoietic Stem/Progenitor Cells via a NOD1-Dependent Pathway in db/db Mice

Figure 1

Diabetes increased the translocation of the bacteria-derived PAMP peptidoglycan and impaired HSPCs in the bone marrow. (a, b) ELISA was used to determine PGN levels in both the (a) plasma and (b) bone marrow extracellular fluid in diabetic db/db mice ( per group). (c) Representative gating scheme for the multicolor flow cytometry analysis of HSPC subpopulations. (d–f) The percentages of (d) LSK (Lin^-CD127^-Sca1⁺c-Kit⁺), (e) LT-HSC (Lin^-CD127^-Sca1⁺c-Kit⁺CD34^-CD135^-), and (f) ST-HSC (Lin^-CD127^-Sca1⁺c-Kit⁺CD34⁺CD135^-) out of total live cells in the bone marrow from db/db mice that had diabetes for 4 months and the age-matched db/m control mice ( per group). (g) ACS-lysed cells from bone marrow or blood were plated in MethoCult GF M3434 medium for the CFU assay ( per group). (h) sorted LSK cells from db/m and db/db mice were cultured for 4 weeks and then plated in MethoCult GF M3434 in triplicate and tested for colonies in the LTC-IC assay ( per group). (i) The percentage of BrdU⁺CD34^-LSK cells was analyzed by FACS after 2-day BrdU administration in BrdU incorporation assay ( per group). Data represent . PGN: peptidoglycan; BM: bone marrow; LT-HSC: long-term repopulating hematopoietic stem cell; ST-HSC: short-term repopulating hematopoietic stem cell. , .

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