Cellular and Molecular Neuroscience, National Brain Research Center, Manesar, Gurgaon, Haryana, India.
PLoS One. 2011 Mar 24;6(3):e18116. doi: 10.1371/journal.pone.0018116.
The neuropathological abnormalities of human immunodeficiency virus (HIV)-1 patients abusing illicit drugs suggest extensive interactions between the two agents, thereby leading to increased rate of progression to neurodegeneration. The role of HIV-1 transactivating protein, Tat has been elucidated in mediating neuronal damage via apoptosis, a hallmark of HIV-associated dementia (HAD), however the underlying mechanisms involved in enhanced neurodegeneration by illicit drugs remain elusive. In this study, we demonstrated that morphine enhances HIV-Tat induced toxicity in human neurons and neuroblastoma cells. Enhanced toxicity by Tat and morphine was accompanied by increased numbers of TUNEL positive apoptotic neurons, elevated caspase-3 levels and decreased ratio of anti- and pro-apoptotic proteins, Bcl2/Bax. Tat and morphine together elicited high levels of reactive oxygen species that were NADPH dependent. Significant alterations in mitochondrial membrane homeostasis were also observed with co-exposure of these agents. Extensive studies of mitogen activated protein kinase (MAPK) signaling pathways revealed the involvement of c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase-1/2 (ERK1/2) pathways in enhanced toxicity of Tat and morphine. In addition to this, we found that pre-treatment of cells with platelet derived growth factor (PDGF-BB) protected neurons from HIV-Tat and morphine induced damage. PDGF-BB alleviated ROS production, maintained mitochondrial membrane potential, decreased caspase-3 activation and hence protected the cells from undergoing apoptosis. PDGF-BB mediated protection against Tat and morphine involved the phosphatidylinositol-3 kinase (PI3K) pathway, as specific inhibitor of PI3K abrogated the protection conferred by PDGF-BB. This study demonstrates the mechanism of enhanced toxicity in human neurons subjected to co-exposure of HIV protein Tat and morphine, thus implying its importance in HIV positive drug abusers, where damage to the brain is reported to be more severe than non-drug abusers. We have also showed for the first time that PDGF-BB can protect against simultaneous exposure of Tat and morphine, strengthening its role as a neuroprotective agent that could be considered for therapeutic intervention.
人类免疫缺陷病毒(HIV)-1 患者滥用非法药物导致的神经病理学异常表明,这两种物质之间存在广泛的相互作用,从而导致神经退行性变的进展速度加快。HIV-1 转录激活蛋白 Tat 在介导通过细胞凋亡导致的神经元损伤方面的作用已得到阐明,细胞凋亡是 HIV 相关痴呆(HAD)的标志,但是,非法药物增强神经退行性变的潜在机制仍难以捉摸。在这项研究中,我们证明了吗啡增强了 HIV-Tat 诱导的人类神经元和神经母细胞瘤细胞毒性。Tat 和吗啡增强的毒性伴随着 TUNEL 阳性凋亡神经元数量的增加、caspase-3 水平的升高以及抗凋亡蛋白和促凋亡蛋白 Bcl2/Bax 的比例降低。Tat 和吗啡共同引起的活性氧水平很高,并且依赖于 NADPH。这些药物共同暴露也观察到线粒体膜稳态的重大改变。对有丝分裂原激活的蛋白激酶(MAPK)信号通路的广泛研究表明,c-Jun N-末端激酶(JNK)和细胞外信号调节激酶-1/2(ERK1/2)通路参与了 Tat 和吗啡增强的毒性。除此之外,我们发现细胞预先用血小板衍生生长因子(PDGF-BB)处理可保护神经元免受 HIV-Tat 和吗啡诱导的损伤。PDGF-BB 减轻了 ROS 的产生,维持了线粒体膜电位,降低了 caspase-3 的激活,从而使细胞免于凋亡。PDGF-BB 对 Tat 和吗啡的保护作用涉及磷脂酰肌醇-3 激酶(PI3K)途径,因为 PI3K 的特异性抑制剂消除了 PDGF-BB 赋予的保护作用。这项研究证明了人类神经元在同时暴露于 HIV 蛋白 Tat 和吗啡时毒性增强的机制,这表明其在 HIV 阳性药物滥用者中具有重要意义,据报道,与非药物滥用者相比,此类人群的大脑损伤更为严重。我们还首次表明,PDGF-BB 可以防止 Tat 和吗啡同时暴露,从而增强了其作为神经保护剂的作用,这可以考虑用于治疗干预。