Wang Ming, Xu Binyuan, Xie Yangmei, Yao Ge, Chen Yinghui
Department of Neurology, Huashan Hospital, Fudan University, Shanghai, 200040, China.
Department of Neurology, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200092, China.
Neurochem Res. 2024 Jul;49(7):1782-1793. doi: 10.1007/s11064-024-04131-x. Epub 2024 Mar 30.
Convulsive status epilepticus (CSE) is a common critical neurological condition that can lead to irreversible hippocampal neuron damage and cognitive dysfunction. Multiple studies have demonstrated the critical roles that long non-coding RNA Mir155hg plays in a variety of diseases. However, less is known about the function and mechanism of Mir155hg in CSE. Here we investigate and elucidate the mechanism underlying the contribution of Mir155hg to CSE-induced hippocampal neuron injury. By applying high-throughput sequencing, we examined the expression of differentially expressed genes in normal and CSE rats. Subsequent RT-qPCR enabled us to measure the level of Mir155hg in rat hippocampal tissue. Targeted knockdown of Mir155hg was achieved by the AAV9 virus. Additionally, we utilized HE and Tunel staining to evaluate neuronal injury. Immunofluorescence (IF), Golgi staining, and brain path clamping were also used to detect the synaptic plasticity of hippocampal neurons. Finally, through IF staining and Sholl analysis, we assessed the degree of microglial phagocytic function. It was found that the expression of Mir155hg was elevated in CSE rats. HE and Tunel staining results showed that Mir155hg knockdown suppressed the hippocampal neuron loss and apoptosis followed CSE. IF, Golgi staining and brain path clamp data found that Mir155hg knockdown enhanced neuronal synaptic plasticity. The results from IF staining and Sholl analysis showed that Mir155hg knockdown enhanced microglial phagocytosis. Our findings suggest that Mir155hg promotes CSE-induced hippocampal neuron injury by inhibiting microglial phagocytosis.
惊厥性癫痫持续状态（CSE）是一种常见的严重神经系统疾病，可导致不可逆的海马神经元损伤和认知功能障碍。多项研究表明，长链非编码RNA Mir155hg在多种疾病中发挥关键作用。然而，关于Mir155hg在CSE中的功能和机制知之甚少。在此，我们研究并阐明Mir155hg对CSE诱导的海马神经元损伤作用的潜在机制。通过高通量测序，我们检测了正常大鼠和CSE大鼠中差异表达基因的表达情况。随后的RT-qPCR使我们能够测量大鼠海马组织中Mir155hg的水平。通过AAV9病毒实现对Mir155hg的靶向敲低。此外，我们利用苏木精-伊红（HE）染色和TUNEL染色评估神经元损伤。免疫荧光（IF）、高尔基染色和脑片钳也用于检测海马神经元的突触可塑性。最后，通过IF染色和Sholl分析，我们评估了小胶质细胞吞噬功能的程度。结果发现，CSE大鼠中Mir155hg的表达升高。HE染色和TUNEL染色结果显示，敲低Mir155hg可抑制CSE后海马神经元的丢失和凋亡。IF染色、高尔基染色和脑片钳数据表明，敲低Mir155hg可增强神经元的突触可塑性。IF染色和Sholl分析结果显示，敲低Mir155hg可增强小胶质细胞的吞噬作用。我们的研究结果表明，Mir155hg通过抑制小胶质细胞吞噬作用促进CSE诱导的海马神经元损伤。
