Neuron-Glia Crosstalk Plays a Major Role in the Neurotoxic Effects of Ketamine via Extracellular Vesicles.
Neuron-Glia Crosstalk Plays a Major Role in the Neurotoxic Effects of Ketamine via Extracellular Vesicles.
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DOI：
10.3389/fcell.2021.691648
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发表时间：
2021
影响因子：
5.5
通讯作者：
Brodie C
中科院分区：
文献类型：
--
作者：
Penning DH;Cazacu S;Brodie A;Jevtovic-Todorovic V;Kalkanis SN;Lewis M;Brodie C
背景：来自动物模型的令人信服的证据表明，早期暴露于临床相关的全身麻醉剂（GAs）会干扰大脑发育，导致长期的认知障碍。由于许多混杂因素，人体研究尚无定论，而且具有挑战性。在这里，我们利用原代人神经细胞来分析氯胺酮的神经毒性作用，重点关注神经胶质细胞的作用及其激活状态。我们还探讨了星形胶质细胞衍生的细胞外囊泡（EVs）和脑源性神经营养因子（BDNF）途径的不同组成部分的作用。方法：采用活/死实验、caspase 3活性和PARP-1裂解率分析氯胺酮对细胞死亡的影响。采用RT-PCR、ELISA和吞噬实验检测星形细胞和小胶质细胞的分化情况。使用transwell培养分析了氯胺酮神经毒性作用中神经元-胶质细胞相互作用的影响。此外，还研究了分离ev和分泌ev在串扰中的作用。采用ELISA、RT-PCR和基因沉默等方法分析BDNF通路不同组分的表达和功能。结果：氯胺酮诱导神经元和少突胶质细胞凋亡，促进促炎星形胶质细胞（A1）和小胶质细胞（M1）表型。星形胶质细胞和小胶质细胞增强了氯胺酮对神经元细胞的神经毒性作用，而神经元增加了少突胶质细胞的细胞死亡。氯胺酮调节BDNF通路的不同组分：降低神经元和星形胶质细胞中BDNF的分泌，同时增加神经元中p75的表达以及神经元和星形胶质细胞中BDNF- as和pro-BDNF的分泌。我们证明了氯胺酮处理的星形胶质细胞分泌的ev在神经元细胞死亡中的重要作用，以及ev相关的BDNF-AS在这一作用中的作用。结论：氯胺酮对神经细胞和非神经细胞均有神经毒性作用。BDNF通路和星形胶质细胞衍生的ev是氯胺酮作用的重要介质。这些结果有助于更好地了解氯胺酮对人类的神经毒性作用，并有助于开发潜在的方法来减少其对神经发育的影响。
Background: There is a compelling evidence from animal models that early exposure to clinically relevant general anesthetics (GAs) interferes with brain development, resulting in long-lasting cognitive impairments. Human studies have been inconclusive and are challenging due to numerous confounding factors. Here, we employed primary human neural cells to analyze ketamine neurotoxic effects focusing on the role of glial cells and their activation state. We also explored the roles of astrocyte-derived extracellular vesicles (EVs) and different components of the brain-derived neurotrophic factor (BDNF) pathway. Methods: Ketamine effects on cell death were analyzed using live/dead assay, caspase 3 activity and PARP-1 cleavage. Astrocytic and microglial cell differentiation was determined using RT-PCR, ELISA and phagocytosis assay. The impact of the neuron-glial cell interactions in the neurotoxic effects of ketamine was analyzed using transwell cultures. In addition, the role of isolated and secreted EVs in this cross-talk were studied. The expression and function of different components of the BDNF pathway were analyzed using ELISA, RT-PCR and gene silencing. Results: Ketamine induced neuronal and oligodendrocytic cell apoptosis and promoted pro-inflammatory astrocyte (A1) and microglia (M1) phenotypes. Astrocytes and microglia enhanced the neurotoxic effects of ketamine on neuronal cells, whereas neurons increased oligodendrocyte cell death. Ketamine modulated different components in the BDNF pathway: decreasing BDNF secretion in neurons and astrocytes while increasing the expression of p75 in neurons and that of BDNF-AS and pro-BDNF secretion in both neurons and astrocytes. We demonstrated an important role of EVs secreted by ketamine-treated astrocytes in neuronal cell death and a role for EV-associated BDNF-AS in this effect. Conclusions: Ketamine exerted a neurotoxic effect on neural cells by impacting both neuronal and non-neuronal cells. The BDNF pathway and astrocyte-derived EVs represent important mediators of ketamine effects. These results contribute to a better understanding of ketamine neurotoxic effects in humans and to the development of potential approaches to decrease its neurodevelopmental impact.
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DOI：
10.1016/s0140-6736(15)00608-x
发表时间：
2016-01-16
期刊：
Lancet (London, England)
影响因子：
--
作者：
Davidson AJ;Disma N;de Graaff JC;Withington DE;Dorris L;Bell G;Stargatt R;Bellinger DC;Schuster T;Arnup SJ;Hardy P;Hunt RW;Takagi MJ;Giribaldi G;Hartmann PL;Salvo I;Morton NS;von Ungern Sternberg BS;Locatelli BG;Wilton N;Lynn A;Thomas JJ;Polaner D;Bagshaw O;Szmuk P;Absalom AR;Frawley G;Berde C;Ormond GD;Marmor J;McCann ME;GAS consortium
通讯作者：
GAS consortium
影响因子：
--
作者：
Jiang W;Finniss S;Cazacu S;Xiang C;Brodie Z;Mikkelsen T;Poisson L;Shackelford DB;Brodie C
通讯作者：
Brodie C
影响因子：
14
作者：
Bier, Ariel;Berenstein, Peter;Brodie, Chaya
通讯作者：
Brodie, Chaya
影响因子：
6.2
作者：
Chaudhuri, Amrita Datta;Dasgheyb, Raha M.;Haughey, Norman J.
通讯作者：
Haughey, Norman J.
影响因子：
9
作者：
Bier A;Hong X;Cazacu S;Goldstein H;Rand D;Xiang C;Jiang W;Ben-Asher HW;Attia M;Brodie A;She R;Poisson LM;Brodie C
通讯作者：
Brodie C