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

Abstract

Critical limb ischemia (CLI) is a severe complication of diabetes mellitus that occurs without effective therapy. Excessive reactive oxygen species (ROS) production and oxidative stress play critical roles in the development of diabetic cardiovascular complications. N-acetylcysteine (NAC) reduces ischemia-induced ROS production. The present study aimed to investigate the effect of NAC on the recovery of ischemic limb in an experimental model of type-2 diabetes. TALLYHO/JngJ diabetic and SWR/J non-diabetic mice were used for developing a CLI model. For NAC treatment, mice received NAC (1 mg/mL) in their drinking water for 24 h before initiating CLI, and continuously for the duration of the experiment. Blood flow, mechanical function, histology, expression of antioxidant enzymes including superoxide dismutase (SOD)-1, SOD-3, glutathione peroxidase (Gpx)-1, catalase, and phosphorylated insulin receptor substrate (IRS)-1, Akt, and eNOS in ischemic limb were evaluated in vivo or ex vivo. Body weight, blood glucose, plasma advanced glycation end-products (AGEs), plasma insulin, insulin resistance index, and plasma TNF-a were also evaluated during the experiment. NAC treatment effectively attenuated ROS production with preserved expressions of SOD-1, Gpx-1, catalase, phosphorylated Akt, and eNOS, and enhanced the recovery of blood flow and function of the diabetic ischemic limb. NAC treatment also significantly decreased the levels of phosphorylated IRS-1 (Ser307) expression and plasma TNF-α in diabetic mice without significant changes in blood glucose and AGEs levels. In conclusion, NAC treatment enhanced the recovery of blood flow and mechanical function in ischemic limbs in T2D mice in association with improved tissue redox/inflammatory status and insulin resistance.

Co-Delivery of Bioengineered Exosomes and Oxygen for Treating Critical Limb Ischemia in Diabetic Mice

“…It is also possible that a higher dosage of oxygen-releasing nanoparticles will increase deteriorative oxidative stress in the tissues. Third, this study used a well-established mouse CLI model, − where the limb ischemia was induced by ligating at both the proximal and distal portions of the left femoral artery and vein and completely cutting the vessels between the two sutures. The total loss of blood flow between the two sutures quickly develops an ischemic environment.…”

“…Critical limb ischemia surgery was performed on female db/db mice aged 8–10 weeks (Jackson Laboratory). Under anesthesia with 2% isoflurane, one hindlimb of each mouse was ligated with a 6.0 silk thread at both the proximal and distal portions of the left femoral artery and vein, and the vessels were completely severed between the two sutures. − The contralateral hindlimb served as a control. 4 h after the surgery, the mice were randomly divided into five groups ( n ≥ 6 for each group), each of which was intravenously injected with 200 μL of PBS (PBS group), 10 mg/mL of empty nanoparticles in PBS (NP group), 0.3 mg/mL of ILTEXO in PBS (ILTEXO group), 10 mg/mL of ILTONP in PBS (ILTONP group), or ILTEXO and ILTONP (ILTEXO/ILTONP group).…”

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

“…Gastrocnemius muscle was collected from the ischemic limb and prepared ( via homogenization and lysis) for western blot analysis as detailed previously ( 26 ). The lysates of the muscle tissues were placed on 8% SDS-PAGE gels.…”

The effects of advanced glycation end‐products on skin and potential anti‐glycation strategies

“…The antioxidant, such as N-acetylcysteine (NAC), Vitamin C, Vitamin E, can scavenge ROS, prevent oxidative stress and damage. 69,81,82 These are all important approaches to inhibit AGEs.…”