Department of Anesthesiology, Shanghai Children's Medical Center, Shanghai Jiao Tong University School of Medicine, Shanghai, 200127, China; Center for Brain Science, Shanghai Children's Medical Center, Shanghai Jiao Tong University School of Medicine, Shanghai 200127, China.
Center for Brain Science, Shanghai Children's Medical Center, Shanghai Jiao Tong University School of Medicine, Shanghai 200127, China; Department of Pharmacy, Shanghai Children's Medical Center, Shanghai Jiao Tong University School of Medicine, Shanghai 200127, China.
Exp Neurol. 2021 Jun;340:113659. doi: 10.1016/j.expneurol.2021.113659. Epub 2021 Feb 26.
Remodeling of the extracellular matrix (ECM) by matrix metalloproteinases (MMPs) plays a pivotal role for microglia in developing retina. We tested whether integrin-dependent microgliosis mediates ketamine-induced neuronal apoptosis in the developing rat retina.
We performed immunofluorescence assays to investigate the role of integrin receptors expressed in the microglia in ketamine-induced neuronal apoptosis. Quantitative reverse transcription polymerase chain reaction (RT-qPCR) and enzyme-linked immunosorbent assay (ELISA) were used to investigate the protein and mRNA levels of cytokines (TNF-α, IL-1β) and/or chemokines (CCL2, CXCL6, CXCL10, and CXCL12). Experiments were performed using whole-mount retinas dissected from P7 Sprague-Dawley rats.
Integrin receptors expressed in microglia were upregulated in ketamine-induced neuronal apoptosis in the early developing rat retina. Downregulating integrin receptors with RGD peptide ameliorated ketamine-induced microgliosis through: 1) ameliorating the change in microglia morphology from immature ramified microglia to an amoeboid state; 2) decreasing the number of microglia and intensity of activated microglia in the retinal ganglion cell layer (GCL); and 3) decreasing cytokine (TNF-α and IL-1β) and chemokine (CCL2, CXCL10) levels in the retinal tissue. Inhibition of activated microglia with minocycline or the blockade of cytokines (TNF-α and IL-1β) with a receptor antagonist (RA) attenuated neuronal apoptosis after exposure to ketamine.
The upregulation of integrin β1 receptors in the microglia acts as a signaling molecule, triggering microgliosis to aggravate ketamine-induced neuronal apoptosis via the release of TNF-α and IL-1β in the early developing rat retina.
细胞外基质（ECM）的重塑是由基质金属蛋白酶（MMPs）介导的，这对于发育中的视网膜中的小胶质细胞起着关键作用。我们测试了整合素依赖性小胶质细胞增生是否介导了氯胺酮诱导的发育中大鼠视网膜神经元凋亡。
我们进行免疫荧光检测，以研究小胶质细胞中表达的整合素受体在氯胺酮诱导的神经元凋亡中的作用。定量逆转录聚合酶链反应（RT-qPCR）和酶联免疫吸附试验（ELISA）用于检测细胞因子（TNF-α、IL-1β）和/或趋化因子（CCL2、CXCL6、CXCL10 和 CXCL12）的蛋白和 mRNA 水平。实验使用从 P7 斯普拉格-道利大鼠分离的全视网膜进行。
在早期发育的大鼠视网膜中，氯胺酮诱导的神经元凋亡中小胶质细胞中表达的整合素受体上调。用 RGD 肽下调整合素受体通过以下方式改善氯胺酮诱导的小胶质细胞增生：1）改善小胶质细胞形态从未成熟的分枝状小胶质细胞向阿米巴样状态的变化；2）减少视网膜神经节细胞层（GCL）中小胶质细胞的数量和活化小胶质细胞的强度；3）降低视网膜组织中的细胞因子（TNF-α和 IL-1β）和趋化因子（CCL2、CXCL10）水平。用米诺环素抑制活化的小胶质细胞或用受体拮抗剂（RA）阻断细胞因子（TNF-α和 IL-1β），可减轻氯胺酮暴露后神经元的凋亡。
小胶质细胞中整合素β1 受体的上调作为一种信号分子，通过释放 TNF-α和 IL-1β，在早期发育的大鼠视网膜中触发小胶质细胞增生，加重氯胺酮诱导的神经元凋亡。
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