HIV-1 Tat enhances purinergic P2Y4 receptor signaling to mediate inflammatory cytokine production and neuronal damage via PI3K/Akt and ERK MAPK pathways
HIV-1 Tat enhances purinergic P2Y4 receptor signaling to mediate inflammatory cytokine production and neuronal damage via PI3K/Akt and ERK MAPK pathways
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HIV-1 Tat 增强嘌呤能 P2Y4 受体信号传导，通过 PI3K/Akt 和 ERK MAPK 途径介导炎症细胞因子的产生和神经元损伤
DOI：
10.1186/s12974-019-1466-8
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发表时间：
2019
影响因子：
9.3
通讯作者：
Gao Dianshuai
中科院分区：
文献类型：
--
作者：
Zhou Feng;Liu Xiaomei;Gao Lin;Zhou Xinxin;Cao Qianwen;Niu Liping;Wang Jing;Zuo Dongjiao;Li Xiangyang;Yang Ying;Hu Minmin;Yu Yinghua;Tang Renxian;Lee Bong Ho;Choi Byoung Wook;Wang Yugang;Izumiya Yoshihiro;Xue Min;Zheng Kuiyang;Gao Dianshuai
研究背景HIV相关的神经认知障碍（HANDs）困扰着半数以上的HIV-1阳性个体。HIV病毒产生的转录反式激活因子（transactivator of transcription，达特）参与炎症反应，是HAND发病过程中引起神经元损伤的主要神经毒性介质。活化的星形胶质细胞是参与神经炎症和神经元损伤的重要细胞。星形胶质细胞中表达的嘌呤能受体参与病毒诱导的神经毒性的正反馈回路。在这里，我们调查，是否P2 Y受体亚型，在星形胶质细胞中表达的参与在体外和体内Tat诱导的神经元death.Methodssoluble达特蛋白进行，以确定表达的P2 Y 4 R和促炎细胞因子在星形胶质细胞中使用siRNA技术，通过实时PCR，Western blot和免疫荧光测定。使用流式细胞仪珠阵列测量促炎细胞因子释放。采用TUNEL染色和MTT细胞活力法检测HT 22细胞的凋亡和存活率，ApopTag®过氧化物酶原位凋亡检测试剂盒和甲酚紫染色检测海马神经元的凋亡和死亡。星形胶质细胞中的P2 Y 4 R信号通过PI 3 K/Akt和ERK 1/2依赖性途径参与Tat诱导的炎性细胞因子产生。敲低P2 Y 4 R表达显著减少炎性细胞因子的产生，并减轻体外Tat介导的神经元凋亡。此外，在体内挑战与达特，P2 Y 4 R基因敲除小鼠表现出减少炎症和神经元损伤，特别是在海马CA 1 region.ConclusionsOur数据提供了新的见解星形胶质细胞介导的神经元损伤在HIV-1感染，并提出了一个潜在的治疗目标的手。
BackgroundHIV-associated neurocognitive disorders (HANDs) afflict more than half of HIV-1-positive individuals. The transactivator of transcription (Tat) produced by HIV virus elicits inflammatory process and is a major neurotoxic mediator that induce neuron damage during HAND pathogenesis. Activated astrocytes are important cells involved in neuroinflammation and neuronal damage. Purinergic receptors expressed in astrocytes participate in a positive feedback loop in virus-induced neurotoxicity. Here, we investigated that whether P2Y4R, a P2Y receptor subtype, that expressed in astrocyte participates in Tat-induced neuronal death in vitro and in vivo.MethodsSoluble Tat protein was performed to determine the expression of P2Y4R and proinflammatory cytokines in astrocytes using siRNA technique via real-time PCR, Western blot, and immunofluorescence assays. Cytometric bead array was used to measure proinflammatory cytokine release. The TUNEL staining and MTT cell viability assay were analyzed for HT22 cell apoptosis and viability, and the ApopTag® peroxidase in situ apoptosis detection kit and cresyl violet staining for apoptosis and death of hippocampal neuron in vivo.ResultsWe found that Tat challenge increased the expression of P2Y4R in astrocytes. P2Y4R signaling in astrocytes was involved in Tat-induced inflammatory cytokine production via PI3K/Akt- and ERK1/2-dependent pathways. Knockdown of P2Y4R expression significantly reduced inflammatory cytokine production and relieved Tat-mediated neuronal apoptosis in vitro. Furthermore, in vivo challenged with Tat, P2Y4R knockdown mice showed decreased inflammation and neuronal damage, especially in hippocampal CA1 region.ConclusionsOur data provide novel insights into astrocyte-mediated neuron damage during HIV-1 infection and suggest a potential therapeutic target for HANDs.