Effects Of N-acetylcysteine Amide Research Articles

The Role of N-Acetylcysteine Amide (NACA) As Prophylactic Treatment for Acute Graft-Versus-Host Disease

Aim The current study aimed to investigate the role of N-acetylcysteine amide (NACA) as prophylactic treatment for acute graft-versus-host disease (GvHD) and to explore the underlying mechanism of action. Background GvHD remains one of the major complications after allogeneic hematopoietic cell transplantation (allo-HCT). In addition, GvHD is a significant cause of morbidity, mortality and deteriorated life quality in transplanted patients. At the present, several strategies have been suggested as a prophylactic treatment against GvHD. Due to the good anti-proliferative effects on the donor T cells, calcineurin inhibitors (cyclosporine or tacrolimus) in combination with methotrexate (MTX) or mycophenolate mofetil (MMF) are recommended by several transplantation centers. Recently, several studies have reported good results of post-transplant cyclophosphamide treatment because of its ability to induce apoptosis of donor allo-activated T cells and to promote transplant tolerance. However, inevitable off-target effect and dose-related toxicity frequently occur regardless of the treatment selected. Evidence showed that oxidative stress is elevated in the recipients of allo-HCT and is linked to the pathogenesis of GvHD. Methods Using a classic murine GvHD model with complete MHC-mismatched allo-HCT (C57Bl/6 to Balb/c), the effect of NACA on the occurrence and severity of acute GvHD was evaluated. In order to reveal the underling mechanism of action, clinical manifestations, survival, cytokine profiles, ROS accumulation, T cell allo-responses, and target tissue damage in NACA-treated GvHD mice were scrutinized and compared with the same parameters observed in N-acetylcysteine (NAC)-treated GvHD mice. Results We found that NACA treatment significantly prolonged the survival and decreased GvHD severity score, in mice underwent allo-HCT. Moreover, NACA downregulated the pro-inflammatory cytokines, such as IFNγ, TNFα, IL-1β and IL-6, but upregulated the anti-inflammatory IL-4 in the mouse serum. Data also showed that NACA administration significantly reduced ROS accumulation in the spleen on day +7 and day +14 after transplantation. In addition, a modest dividing defect in donor T cells was observed upon NACA administration regardless of the experienced antigen classes, and the remarkably lower percentage of Ki67 expression in donor T cells substantiated their inhibited proliferation. In the meanwhile, NACA restrained Th1 and Th17 differentiation of donor T cells but enhanced the regulatory polarization. Furthermore, the migration capability of donor T cells was suppressed by NACA as well, resulting in a less severe intervention of the GvHD target organs. Importantly, NACA did not alter the engraftment as was observed in the bone marrow and spleen chimera but facilitated the neutrophil engraftment. Opposite to NACA, the effect of NAC on GvHD was not significant. Conclusion The present investigation showed that NACA, rather than NAC, has a prophylactic effect in acute GvHD mouse model.The protective effect is mediated by antioxidation, anti-inflammation and downregulation of T-cell allo-responses without altering the engraftment and its process.NACA treatment has significantly improved survival, lower the severity score and shortened the recovery time. However, it was not able to fully eliminate the occurrence of acute GvHD.It is important to consider the combination of NACA with other clinical-used drugs in order to achieve complete disease remission and minimize treatment-related toxicity.The encouraging present results in combination with the clinical reported results of the use of NACA may open the door for a new prophylactic treatment of GvHD.

Effects of N-acetylcysteine amide on anxiety and stress behavior in zebrafish.

Anxiety disorders are highly prevalent and a leading cause of disability worldwide. Their etiology is related to stress, an adaptive response of the organism to restore homeostasis, in which oxidative stress and glutamatergic hyperactivity are involved. N-Acetylcysteine (NAC) is a multitarget approved drug proved to be beneficial in the treatment of various mental disorders. Nevertheless, NAC has low membrane permeability and poor bioavailability and its limited delivery to the brain may explain inconsistencies in the literature. N-Acetylcysteine amide (AD4) is a synthetic derivative of NAC in which the carboxyl group was modified to an amide. The amidation of AD4 improved lipophilicity and blood-brain barrier permeability and enhanced its antioxidant properties. The purpose of this study was to investigate the effects of AD4 on behavioral and biochemical parameters in zebrafish anxiety models. Neither AD4 nor NAC induced effects on locomotion and anxiety-related parameters in the novel tank test. However, in the light/dark test, AD4 (0.001 mg/L) increased the time spent in the lit side in a concentration 100 times lower than NAC (0.1 mg/L). In the acute restraint stress protocol, NAC and AD4 (0.001 mg/L) showed anxiolytic properties without meaningful effects on oxidative status. The study suggests that AD4 has anxiolytic effects in zebrafish with higher potency than the parent compound. Additional studies are warranted to characterize the anxiolytic profile of AD4 and its potential in the management of anxiety disorders.

N-Acetylcysteine Amide (NACA) Reduces Cell Death after Oxidative Stress in a Porcine Embryonic Kidney Cell Line

Introduction: Oxidative stress may have detrimental effects on different structures of the cells, such as the DNA. Recently, we have published a study demonstrating that N-Acetylcysteine amide (NACA) displayed anti-inflammatory properties on the brain after exposure to oxidative stress in an established neonatal piglet model, imitating perinatal asphyxia. As different clinical studies have shown an association between the severity of hypoxic-ischemic encephalopathy and damage of the kidneys, we investigated a possible protective effect of NACA against H2O2-induced oxidative stress using a porcine epithelial-like embryonic kidney cell line (EFN-R). Objective: To investigate a potential protective effect of NACA on cells of a porcine embryonic kidney cell line exposed to H2O2. Methods: We subjected the cells to different concentrations of H2O2 for variable time periods, seeking the optimal dose-response for the experiments. Based on the results of these investigations, we exposed the cells to 100 μMol of H2O2 and/or 750 μM of NACA for 24 hours. Some of the cells would receive NACA either one hour before or one hour after exposure to H2O2. Results: The viability of the investigated EFN-R cells revealed that both, the group treated with NACA before exposure to H2O2 and the group treated with NACA after exposure to H2O2, exhibited significantly higher cell viability compared to the H2O2 group (p < 0.001 and p < 0.01, respectively). Discussion: The increased viability of the cells may indicate that NACA could play an important role in reducing oxidative stress. Taking the results from our previous study into consideration, our findings may strengthen the theory that NACA may have organ protective properties for neonates exposed to oxidative stress.

PSS290 - Effects of N-acetylcysteine Amide (NACA) Eye Drops on Acetaminophen-Induced Cataracts in C57BL/6 Mice

PSS290 - Effects of N-acetylcysteine Amide (NACA) Eye Drops on Acetaminophen-Induced Cataracts in C57BL/6 Mice

In vivo inhibition of l-buthionine-( S,R)-sulfoximine-induced cataracts by a novel antioxidant, N-acetylcysteine amide

The effects of N-acetylcysteine amide (NACA), a free radical scavenger, on cataract development were evaluated in Wistar rat pups. Cataract formation was induced in these animals with an intraperitoneal injection of a glutathione (GSH) synthesis inhibitor, l-buthionine-( S,R)-sulfoximine (BSO). To assess whether NACA has a significant impact on BSO-induced cataracts, the rats were divided into four groups: (1) control, (2) BSO only, (3) NACA only, and (4) NACA + BSO. The control group received only saline ip injections on postpartum day 3, the BSO-only group was given ip injections of BSO (4 mmol/kg body wt), the NACA-only group received ip injections of only NACA (250 mg/kg body wt), and the NACA + BSO group was given a dose of NACA 30 min before administration of the BSO injection. The pups were sacrificed on postpartum day 15, after examination under a slit-lamp microscope. Their lenses were analyzed for selective oxidative stress parameters, including glutathione (reduced and oxidized), protein carbonyls, catalase, glutathione peroxidase, glutathione reductase, and malondialdehyde. The lenses of pups in both the control and the NACA-only groups were clear, whereas all pups within the BSO-only group developed well-defined cataracts. It was found that supplemental NACA injections during BSO treatment prevented cataract formation in most of the rat pups in the NACA + BSO group. Only 20% of these pups developed cataracts, and the rest retained clear lenses. Further, GSH levels were significantly decreased in the BSO-only treated group, but rats that received NACA injections during BSO treatment had these levels of GSH replenished. Our findings indicate that NACA inhibits cataract formation by limiting protein carbonylation, lipid peroxidation, and redox system components, as well as replenishing antioxidant enzymes.

Effects of N-acetylcysteine amide (NACA), a thiol antioxidant on radiation-induced cytotoxicity in Chinese hamster ovary cells

Ionizing radiation is known to cause tissue damage in biological systems, mainly due to its ability to produce reactive oxygen species (ROS) in cells. Many thiol antioxidants have been used previously as radioprotectors, but their application has been limited by their toxicity. In this investigation, we have explored the possible radioprotective effects of a newly synthesized thiol antioxidant, N-acetylcysteine amide (NACA), in comparison with N-acetylcysteine (NAC), a commonly used antioxidant. Protective effects of NACA and NAC were assessed using Chinese hamster ovary (CHO) cells, irradiated with 6 gray (Gy) radiation. Oxidative stress parameters, including levels of reduced glutathione (GSH), cysteine, malondialdehyde (MDA), and activities of antioxidant enzymes like glutathione peroxidase, glutathione reductase, and catalase, were measured. Results indicate that NACA was capable of restoring GSH levels in irradiated cells in a dose dependent manner. In addition, NACA prevented radiation-induced loss in cell viability. NACA further restored levels of malondialdehyde, caspase-3 activity, and antioxidant enzyme activities to control levels. Although NAC affected cells in a similar manner to NACA, its effects were not as significant. Further, NAC was also found to be cytotoxic to cells at higher concentrations, whereas NACA was non-toxic at similar concentrations. These results suggest that NACA may be able to attenuate radiation-induced cytotoxicity, possibly by its ability to provide thiols to cells.

Effects of N-acetylcysteine amide (NACA), a novel thiol antioxidant against glutamate-induced cytotoxicity in neuronal cell line PC12

Oxidative stress plays an important role in neuronal cell death associated with many different neurodegenerative conditions such as cerebral ischemia and Parkinson's disease. Elevated levels of glutamate are thought to be responsible for CNS disorders through various mechanisms causing oxidative stress induced by a nonreceptor-mediated oxidative pathway which blocks cystine uptake and results in depletion of intracellular glutathione (GSH). The newly designed amide form of N-acetylcysteine (NAC), N-acetylcysteine amide (NACA), was assessed for its ability to protect PC12 cells against oxidative toxicity induced by glutamate. NACA was shown to protect PC12 cells from glutamate (Glu) toxicity, as evaluated by LDH and MTS assays. NACA prevented glutamate-induced intracellular GSH loss. In addition, NACA restored GSH synthesis in a Glu (10 mM) plus buthionine–sulfoximine (BSO) (0.2 mM)-treated group, indicating that the intracellular GSH increase is independent of γ-GSC (γ-glutamylcysteinyl synthetase). The increase in levels of reactive oxygen species (ROS) induced by glutamate was significantly decreased by NACA. Measurement of malondialdehyde (MDA) showed that NACA reduced glutamate-induced elevations in levels of lipid peroxidation by-products. These results demonstrate that NACA can protect PC12 cells against glutamate cytotoxicity by inhibiting lipid peroxidation, and scavenging ROS, thus preserving intracellular GSH.