Probenecid and N-Acetylcysteine Prevent Loss of Intracellular Glutathione and Inhibit Neuronal Death after Mechanical Stretch Injury In Vitro

被引:19
作者
Du, Lina [1 ]
Empey, Philip E. [4 ,5 ]
Ji, Jing [1 ,6 ]
Chao, Honglu [1 ]
Kochanek, Patrick M. [1 ,2 ,3 ,7 ,8 ]
Bayir, Hulya [1 ,2 ,7 ,8 ,9 ]
Clark, Robert S. B. [1 ,2 ,5 ,7 ,8 ]
机构
[1] Univ Pittsburgh, Sch Med, Dept Crit Care Med, Pittsburgh, PA USA
[2] Univ Pittsburgh, Sch Med, Dept Pediat, Pittsburgh, PA 15261 USA
[3] Univ Pittsburgh, Sch Med, Dept Anesthesiol, Pittsburgh, PA USA
[4] Univ Pittsburgh, Sch Med, Clin & Translat Sci Inst, Pittsburgh, PA USA
[5] Univ Pittsburgh, Sch Pharm, Dept Pharm & Therapeut, Pittsburgh, PA 15261 USA
[6] Nanjing Med Univ, Dept Neurosurg, Affiliated Hosp 1, Nanjing, Jiangsu, Peoples R China
[7] Safar Ctr Resuscitat Res, Pittsburgh, PA USA
[8] Childrens Hosp Pittsburgh UPMC, Pittsburgh, PA USA
[9] Univ Pittsburgh, Ctr Free Radical & Antioxidant Hlth, Dept Environm & Occupat Hlth, Pittsburgh, PA USA
关键词
adenosine triphosphate binding cassette transporter C1; gender; multidrug resistance associated protein 1; primary cortical neurons; spinal cord injury; traumatic brain injury; TRAUMATIC BRAIN-INJURY; CORTICAL-NEURONS; MICE; NEUROPROTECTION; APOPTOSIS; INFLAMMASOME; TRANSPORTER; PROTECTS; DAMAGE; RATS;
D O I
10.1089/neu.2015.4342
中图分类号
R4 [临床医学];
学科分类号
1002 ; 100602 ;
摘要
Probenecid and N-acetylcysteine (NAC) can preserve intracellular levels of the vital antioxidant glutathione (GSH) via two distinct biochemical pathways. Probenecid inhibits transporter-mediated GSH efflux and NAC serves as a cysteine donor for GSH synthesis. We hypothesized that probenecid and NAC alone would maintain intracellular GSH concentrations and inhibit neuronal death after traumatic stretch injury, and that the drugs in combination would produce additive effects. Sex-segregated rat primary cortical neurons were treated with probenecid (100M) and NAC (50M), alone and in combination (Pro-NAC), then subjected to mechanical stretch (10s(-1) strain rate, 50% membrane deformation). At 24h, both probenecid and NAC inhibited trauma-induced intracellular GSH depletion, lactate dehydrogenase (LDH) release, and propidium iodide (PI) uptake in both XY- and XX-neurons. Combined Pro-NAC treatment was superior to probenecid or NAC alone in maintenance of intracellular GSH and neuronal death assessed by PI uptake. Interestingly, caspase 3 activity 24h after mechanical trauma was more prominent in XX-neurons, and treatment effects (probenecid, NAC, and Pro-NAC) were observed in XX- but not XY-neurons; however, XY-neurons were ultimately more vulnerable to mechanical stretch-induced injury than their XX counterparts, as was evidenced by more neuronal death detected by LDH release and PI uptake. In addition, after stretch injury in HT22 hippocampal cells, both NAC and probenecid were highly effective at reducing oxidative stress detected by dichlorofluorescein fluorescence. These in vitro data support further testing of this drug combination in models of traumatic neuronal injury in vivo.
引用
收藏
页码:1913 / 1917
页数:5
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