Inhibition of catechol-O-methyl transferase (COMT) by tolcapone restores reductions in microtubule-associated protein 2 (MAP2) and synaptophysin (SYP) following exposure of neuronal cells to neurotropic HIV.
Inhibition of catechol-O-methyl transferase (COMT) by tolcapone restores reductions in microtubule-associated protein 2 (MAP2) and synaptophysin (SYP) following exposure of neuronal cells to neurotropic HIV.
复制标题
DOI：
10.1007/s13365-015-0354-y
复制
发表时间：
2015-10
影响因子：
3.2
通讯作者：
Everall IP
中科院分区：
文献类型：
--
作者：
Lee TT;Chana G;Gorry PR;Ellett A;Bousman CA;Churchill MJ;Gray LR;Everall IP
本研究旨在评估托卡朋对cathecol-O-甲基转移酶（COMT）的抑制是否可以提供针对HIV相关神经退行性作用的神经保护。本研究是在前人工作的基础上进行的，前人的工作表明，COMT基因158位（val 158 met）的单核苷酸多态性（SNP）导致COMT活性降低40%。重要的是，这种减少对艾滋病毒相关的神经认知障碍（HAND）具有保护作用，这种疾病与艾滋病毒相关的大脑变化有关。在托卡朋存在或不存在的情况下，将SH-SY 5 Y分化的神经元暴露于巨噬细胞传播的HIV（亲神经性MACS 2-Br株）6天。提取RNA并使用Qiagen RT 2-定制阵列进行qPCR，所述定制阵列由用于神经元和突触完整性的基因、COMT和促炎标志物组成。进行免疫荧光以在蛋白水平验证基因表达变化。我们的研究结果表明，与对照组相比，HIV显著增加COMT的mRNA表达，同时降低微管相关蛋白2（MAP 2）（p=0.0015）和突触素（SYP）（p=0.012）的表达。同时暴露于托卡朋改善了与HIV相关的MAP 2（p=0.009）和COMT（p=0.024）的干扰表达。免疫荧光显示SYP和MAP 2随着暴露于HIV而呈下降趋势，并且与单独的HIV相比，伴随的托卡朋暴露增加了SYP（p=0.016）。我们的研究结果表明，在体外抑制COMT可以改善HIV相关的神经退行性变化，导致结构和突触成分MAP 2和SYP的表达减少。由于HIV相关的树突和突触损伤是HAND的贡献者，因此抑制COMT可能是减轻或预防HAND某些症状的潜在策略。
This investigation aimed to assess whether inhibition of cathecol-O-methyl transferase (COMT) by Tolcapone could provide neuroprotection against HIV-associated neurodegenerative effects. This study was conducted based on previous work, which showed that a single nucleotide polymorphism (SNP) at position 158 (val158met) in COMT, resulted in 40% lower COMT activity. Importantly, this reduction confers a protective effect against HIV-associated neurocognitive disorders (HAND), which have been linked to HIV-associated brain changes. SH-SY5Y differentiated neurons were exposed to macrophage-propagated HIV (neurotropic MACS2-Br strain) in the presence or absence of Tolcapone for 6 days. RNA was extracted and qPCR was performed using Qiagen RT2-custom-array consisting of genes for neuronal and synaptic integrity, COMT and pro-inflammatory markers. Immunofluorescence was conducted to validate the gene expression changes at the protein level. Our findings demonstrated that HIV significantly increased the mRNA expression of COMT while reduced the expression of microtubule-associated protein 2 (MAP2) (p=0.0015) and synaptophysin (SYP) (p=0.012) compared to control. A concomitant exposure of Tolcapone ameliorated the perturbed expression of MAP2 (p=0.009) and COMT (p=0.024) associated with HIV. Immunofluorescence revealed a trend reduction of SYP and MAP2 with exposure to HIV, and that concomitant exposure of Tolcapone increased SYP (p=0.016) compared to HIV alone. Our findings demonstrated in vitro that inhibition of COMT can ameliorate HIV-associated neurodegenerative changes that resulted in the decreased expression of the structural and synaptic components MAP2 and SYP. As HIV-associated dendritic and synaptic damage are contributors to HAND, inhibition of COMT may represent a potential strategy for attenuating or preventing some of the symptoms of HAND.