Neurophysiology Laboratory, Department of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center, Omaha, Nebraska, USA.
PLoS One. 2013 May 30;8(5):e64904. doi: 10.1371/journal.pone.0064904. Print 2013.
Microglia plays a crucial role in the pathogenesis of HIV-1-associated neurocognitive disorders. Increasing evidence indicates the voltage-gated potassium (Kv) channels are involved in the regulation of microglia function, prompting us to hypothesize Kv channels may also be involved in microglia-mediated neurotoxic activity in HIV-1-infected brain. To test this hypothesis, we investigated the involvement of Kv channels in the response of microglia to HIV-1 Tat protein. Treatment of rat microglia with HIV-1 Tat protein (200 ng/ml) resulted in pro-inflammatory microglial activation, as indicated by increases in TNF-α, IL-1β, reactive oxygen species, and nitric oxide, which were accompanied by enhanced outward K(+) current and Kv1.3 channel expression. Suppression of microglial Kv1.3 channel activity, either with Kv1.3 channel blockers Margatoxin, 5-(4-Phenoxybutoxy)psoralen, or broad-spectrum K(+) channel blocker 4-Aminopyridine, or by knockdown of Kv1.3 expression via transfection of microglia with Kv1.3 siRNA, was found to abrogate the neurotoxic activity of microglia resulting from HIV-1 Tat exposure. Furthermore, HIV-1 Tat-induced neuronal apoptosis was attenuated with the application of supernatant collected from K(+) channel blocker-treated microglia. Lastly, the intracellular signaling pathways associated with Kv1.3 were investigated and enhancement of microglial Kv1.3 was found to correspond with an increase in Erk1/2 mitogen-activated protein kinase activation. These data suggest targeting microglial Kv1.3 channels may be a potential new avenue of therapy for inflammation-mediated neurological disorders.
小胶质细胞在 HIV-1 相关神经认知障碍的发病机制中起着至关重要的作用。越来越多的证据表明,电压门控钾 (Kv) 通道参与了小胶质细胞功能的调节,这促使我们假设 Kv 通道也可能参与 HIV-1 感染大脑中小胶质细胞介导的神经毒性活性。为了验证这一假设,我们研究了 Kv 通道是否参与 HIV-1 Tat 蛋白对小胶质细胞的反应。用 HIV-1 Tat 蛋白(200ng/ml)处理大鼠小胶质细胞会导致促炎小胶质细胞激活,表现为 TNF-α、IL-1β、活性氧和一氧化氮增加,同时伴有外向 K(+)电流和 Kv1.3 通道表达增强。用 Kv1.3 通道阻断剂 Margatoxin、5-(4-苯氧基丁氧基)补骨脂素或广谱 K(+)通道阻断剂 4-氨基吡啶,或用 Kv1.3 siRNA 转染小胶质细胞抑制 Kv1.3 表达,发现可以阻断 HIV-1 Tat 暴露引起的小胶质细胞的神经毒性活性。此外,用 K(+)通道阻断剂处理的小胶质细胞上清液应用于 HIV-1 Tat 诱导的神经元凋亡可减轻。最后,研究了与 Kv1.3 相关的细胞内信号通路,发现增强小胶质细胞 Kv1.3 与 Erk1/2 丝裂原激活蛋白激酶的激活增加相对应。这些数据表明,靶向小胶质细胞 Kv1.3 通道可能是治疗炎症介导的神经障碍的一种新的潜在途径。