Abstract 16752: Restoration of Defective Hydrogen Sulfide Production in Bone Marrow Cells From Diabetic Mice Rescues Their Impaired Tissue Reparative Function
Background: Bone marrow cell (BMC)-based treatment for critical limb ischemia (CLI) in diabetic patients yielded a modest therapeutic effect due to cell dysfunction. Therefore, approaches that improve diabetic BMC function may provide therapeutic benefits. Here, we tested the hypotheses that restoration of hydrogen sulfide (H2S) production in diabetic BMCs improves their reparative capacities.
Methods and Results: Both H2S production and cystathionine γ-lyase (CSE), an H2S enzyme, levels were significantly decreased in BMCs from diabetic db/db mice. Administration of H2S donor diallyl trisulfide (DATS) or overexpression of CSE restored H2S production and enhanced cell survival and migratory capacity in high glucose (HG)-treated BMCs. Left hind limb ischemia (HLI) surgery was conducted in db/+ and db/db mice followed by oral administration of DATS and/or local intramuscular injection of GFP-labeled BMCs or CSE-overexpressed BMCs by transfection of GFP lentivirus or RFP-tagged CSE lentivirus (CSE-BMCs). BMCs were isolated from db/db mice. Mice with HLI were divided into six groups: 1) db/+; 2) db/db; 3) db/db+BMCs; 4) db/db+DATS; 5) db/db+DATS+BMCs; 6) db/db+CSE-BMCs. DATS and CSE overexpression greatly enhanced diabetic BMCs retention in ischemic hind limbs (IHL) followed by improved blood perfusion, capillary/arteriole density, skeletal muscle architecture and cell survival, and decreased perivascular CD68+ cell infiltration in IHL of diabetic mice. Interestingly, DATS or CSE overexpression rescued HG-impaired migration, tube formation and survival of BMCs or mature human cardiac microvascular endothelial cells (HCMVECs). Mechanistically, DATS restored nitric oxide production and decreased eNOS-pT495 levels in HCMVECs, and improved BMC angiogenic activity under HG condition. Finally, silencing CSE by siRNA significantly increased eNOS-pT495 levels in HCMVECs.
Conclusions: Decreased CSE-mediated H2S bioavailability is an underlying source of BMC dysfunction in diabetes. Our data indicate that H2S and overexpression of CSE in diabetic BMCs may rescue their dysfunction and open novel avenues for cell-based therapeutics of CLI in diabetic patients.
Author Disclosures: Z. Cheng: None. V. Garikipati: None. E. Nickoloff: None. C. Wang: None. D. Polhemus: None. J. Zhou: None. C. Benedict: None. M. Khan: None. S. Verma: None. J. Rabinowitz: None. D. Lefer: None. R. Kishore: None.
- © 2016 by American Heart Association, Inc.