Department of Neurosurgery, Nanjing Drum Tower Hospital, Affiliated Hospital of Medical School, Nanjing University, Nanjing, China.
Institute of Neurosurgery, Nanjing University, Nanjing, Jiangsu Province, China.
Cell Commun Signal. 2023 Jul 21;21(1):175. doi: 10.1186/s12964-023-01173-6.
The phagocytosis and homeostasis of microglia play an important role in promoting blood clearance and improving prognosis after subarachnoid hemorrhage (SAH). LC3-assocaited phagocytosis (LAP) contributes to the microglial phagocytosis and homeostasis via autophagy-related components. With RNA-seq sequencing, we found potential signal pathways and genes which were important for the LAP of microglia.
We used an in vitro model of oxyhemoglobin exposure as SAH model in the study. RNA-seq sequencing was performed to seek critical signal pathways and genes in regulating LAP. Bioparticles were used to access the phagocytic ability of microglia. Western blot (WB), immunoprecipitation, quantitative polymerase chain reaction (qPCR) and immunofluorescence were performed to detect the expression change of LAP-related components and investigate the potential mechanisms.
In vitro SAH model, there were increased inflammation and decreased phagocytosis in microglia. At the same time, we found that the LAP of microglia was inhibited in all stages. RNA-seq sequencing revealed the importance of P38 MAPK signal pathway and DAPK1 in regulating microglial LAP. P38 was found to regulate the expression of DAPK1, and P38-DAPK1 axis was identified to regulate the LAP and homeostasis of microglia after SAH. Finally, we found that P38-DAPK1 axis regulated expression of BECN1, which indicated the potential mechanism of P38-DAPK1 axis regulating microglial LAP.
P38-DAPK1 axis regulated the LAP of microglia via BECN1, affecting the phagocytosis and homeostasis of microglia in vitro SAH model. Video Abstract.
小胶质细胞的吞噬作用和内稳态在促进蛛网膜下腔出血（SAH）后血液清除和改善预后方面发挥着重要作用。LC3 相关的吞噬作用（LAP）通过自噬相关成分促进小胶质细胞的吞噬作用和内稳态。通过 RNA-seq 测序，我们发现了与小胶质细胞 LAP 相关的潜在信号通路和基因。
我们在研究中使用了氧合血红蛋白暴露的体外模型作为 SAH 模型。进行 RNA-seq 测序以寻找调节 LAP 的关键信号通路和基因。生物颗粒用于评估小胶质细胞的吞噬能力。通过 Western blot（WB）、免疫沉淀、定量聚合酶链反应（qPCR）和免疫荧光检测 LAP 相关成分的表达变化，并探讨潜在的机制。
体外 SAH 模型中小胶质细胞的炎症增加，吞噬作用减少。同时，我们发现小胶质细胞的 LAP 在所有阶段都受到抑制。RNA-seq 测序揭示了 P38 MAPK 信号通路和 DAPK1 在调节小胶质细胞 LAP 中的重要性。发现 P38 调节 DAPK1 的表达，并且 P38-DAPK1 轴被鉴定为调节 SAH 后小胶质细胞的 LAP 和内稳态。最后，我们发现 P38-DAPK1 轴调节 BECN1 的表达，这表明了 P38-DAPK1 轴调节小胶质细胞 LAP 的潜在机制。
P38-DAPK1 轴通过 BECN1 调节小胶质细胞的 LAP，影响体外 SAH 模型中小胶质细胞的吞噬作用和内稳态。视频摘要。
