Combination of Antioxidant Enzyme Overexpression and N‐Acetylcysteine Treatment Enhances the Survival of Bone Marrow Mesenchymal Stromal Cells in Ischemic Limb in Mice With Type 2 Diabetes

Abstract

Background Therapy with mesenchymal stem cells remains a promising but challenging approach to critical limb ischemia in diabetes because of the dismal cell survival. Methods and Results Critical limb ischemia in type 2 diabetes mouse model was used to explore the impact of diabetic limb ischemia on the survival of bone marrow mesenchymal stromal cells (bMSCs). Inhibition of intracellular reactive oxygen species was achieved with concomitant overexpression of superoxide dismutase (SOD)‐1 and glutathione peroxidase‐1 in the transplanted bMSCs, and extracellular reactive oxygen species was attenuated using SOD‐3 overexpression and N‐acetylcysteine treatment. In vivo optical fluorescence imaging and laser Doppler perfusion imaging were used to track cell retention and determine blood flow in diabetic ischemic limb, respectively. Survival of the transplanted bMSCs was significantly decreased in diabetic ischemic limb compared with the control. In vitro study indicated that advanced glycation end products, not high glucose, significantly decreased the proliferation of bMSCs and increased their apoptosis associated with increased reactive oxygen species production and selective reduction of SOD‐1 and SOD‐3. In vivo study demonstrated that concomitant overexpression of SOD‐1, SOD‐3, and glutathione peroxidase‐1, or host treatment with N‐acetylcysteine, significantly enhanced in vivo survival of transplanted bMSCs, and improved critical limb ischemia in diabetic mice. Combination of triple antioxidant enzyme overexpression in bMSCs with host N‐acetylcysteine treatment further improved bMSC survival with enhanced circulatory and functional recovery from diabetic critical limb ischemia. Conclusions Simultaneous suppression of reactive oxygen species from transplanted bMSCs and host tissue could additively enhance bMSC survival in diabetic ischemic limb with increased therapeutic efficacy in diabetes.

N‑acetyl cysteine prevents ambient fine particulate matter‑potentiated atherosclerosis via inhibition of reactive oxygen species‑induced oxidized low density lipoprotein elevation and decreased circulating endothelial progenitor cell

“…NAC prevents atherosclerosis formation via suppression of inflammation by inhibiting NF-κB-induced TNF-α production and blood homocysteine levels ( 55 , 56 ). NAC maintains EPC population, decrease conversion of LDL to ox-LDL in vivo , prevent atherosclerosis in humans and mice with hyperlipidemia and promote hind limb ischemia recovery in mice ( 15 , 23 , 30 , 57 ).…”

CARD9 deficiency improves the recovery of limb ischemia in mice with ambient fine particulate matter exposure

“…There were 4 experimental groups with 7 mice in each group: (1) WT-PBS control (WT C57BL/6 mice with CLI and PBS treatment); (2) WT-PM group (WT C57BL/6 mice with CLI and PM treatment); (3) CARD9-PBS group (CARD9 −/− mice with CLI and PBS treatment); and (4) CARD9-PM group (CARD9 −/− mice with CLI and PM treatment). For surgical induction of CLI, a 3–5 mm incision was made, with minimal tissue disturbance, and the right femoral artery was identified and ligated using a 6-0 silk suture and then transected, under general anesthesia with isoflurane (1.5%) and constant temperature (37 ± 0.5°C) as described ( 18 , 19 ). Successful creation of CLI was confirmed by a lack of femoral artery blood flow signal using Laser Doppler perfusion imaging (LDPI, Moor Instruments, Devon, UK).…”

N-Acetylcysteine Enhances the Recovery of Ischemic Limb in Type-2 Diabetic Mice

“…All the surgical procedures were conducted under air anesthesia (1.25% isoflurane) and constant temperature (37 ± 0.5 °C). For mice undergoing NAC treatment, NAC (Sigma-Aldrich, St. Louis, US) was added to drinking water (1 mg/mL) for 24 h before creation of CLI, and continuously throughout the duration of the experiment, based on our previous studies [ 16 , 24 , 25 ] and those of others [ 26 , 27 ]. Based on the amount of water consumed by the mice, the dose of NAC was estimated to be 0.5–1.0 g/kg/day.…”