GSDMD gene knockout alleviates hyperoxia-induced hippocampal brain injury in neonatal mice.
GSDMD gene knockout alleviates hyperoxia-induced hippocampal brain injury in neonatal mice.
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DOI：
10.1186/s12974-023-02878-8
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发表时间：
2023-09-07
影响因子：
9.3
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中科院分区：
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新生儿高氧暴露与早产儿脑损伤和不良神经发育结局相关。我们以前在新生啮齿动物模型中的研究表明，高氧刺激大脑的炎性体通路，导致gasdermin D（GSDMD）的激活，GSDMD是pyroptotic炎性细胞死亡的关键执行者。此外，我们发现，药物抑制半胱天冬酶-1，阻断GSDMD激活，减轻高氧诱导的新生小鼠脑损伤。我们假设GSDMD在高氧诱导的新生儿脑损伤中起致病作用，并且GSDMD基因敲除（KO）将减轻高氧诱导的脑损伤。新生GSDMD基因敲除小鼠及其野生型（WT）同窝出生后24小时内随机暴露于室内空气或高氧（85%O2）从出生后第1天至第14天。在脑切片中通过免疫组织学检测同种异体移植物炎性因子1（AIF 1）和CD 68（小胶质细胞活化的标志物）来评估海马脑炎性损伤。Ki-67染色检测细胞增殖情况，TUNEL法检测细胞死亡情况。对海马进行RNA测序以鉴定高氧和GSDMD-KO的转录效应，并进行qRT-PCR以确认一些显著调节的基因。高氧暴露WT小鼠的小胶质细胞增加与激活一致，这与海马区细胞增殖减少和细胞死亡增加有关。相反，高氧暴露的GSDMD-KO小鼠表现出相当大的耐高氧性，因为O2暴露不会增加AIF 1+、CD 68+或TUNEL +细胞数量或降低细胞增殖。与暴露于室内空气的WT和GSDMD-KO相比，高氧暴露分别在WT中差异调节258个基因，在GSDMD-KO小鼠中仅调节16个基因。基因集富集分析表明，在WT脑中，高氧差异调节基因与神经元和血管的发育和分化，轴突生成，胶质细胞分化，缺氧诱导因子1途径，神经生长因子途径。GSDMD-KO阻止了这些变化。GSDMD-KO证实了新生小鼠海马中高氧诱导的炎性损伤、细胞存活和死亡以及参与神经元生长、发育和分化的通路的转录基因表达的改变。这表明GSDMD在早产儿脑损伤中起致病作用，靶向GSDMD可能有助于预防和治疗早产儿脑损伤和不良神经发育结局。在线版本包含补充材料，可通过10.1186/s12974-023-02878-8获得。
Neonatal hyperoxia exposure is associated with brain injury and poor neurodevelopment outcomes in preterm infants. Our previous studies in neonatal rodent models have shown that hyperoxia stimulates the brain’s inflammasome pathway, leading to the activation of gasdermin D (GSDMD), a key executor of pyroptotic inflammatory cell death. Moreover, we found pharmacological inhibition of caspase-1, which blocks GSDMD activation, attenuates hyperoxia-induced brain injury in neonatal mice. We hypothesized that GSDMD plays a pathogenic role in hyperoxia-induced neonatal brain injury and that GSDMD gene knockout (KO) will alleviate hyperoxia-induced brain injury. Newborn GSDMD knockout mice and their wildtype (WT) littermates were randomized within 24 h after birth to be exposed to room air or hyperoxia (85% O2) from postnatal days 1 to 14. Hippocampal brain inflammatory injury was assessed in brain sections by immunohistology for allograft inflammatory factor 1 (AIF1) and CD68, markers of microglial activation. Cell proliferation was evaluated by Ki-67 staining, and cell death was determined by TUNEL assay. RNA sequencing of the hippocampus was performed to identify the transcriptional effects of hyperoxia and GSDMD-KO, and qRT-PCR was performed to confirm some of the significantly regulated genes. Hyperoxia-exposed WT mice had increased microglia consistent with activation, which was associated with decreased cell proliferation and increased cell death in the hippocampal area. Conversely, hyperoxia-exposed GSDMD-KO mice exhibited considerable resistance to hyperoxia as O2 exposure did not increase AIF1 + , CD68 + , or TUNEL + cell numbers or decrease cell proliferation. Hyperoxia exposure differentially regulated 258 genes in WT and only 16 in GSDMD-KO mice compared to room air-exposed WT and GSDMD-KO, respectively. Gene set enrichment analysis showed that in the WT brain, hyperoxia differentially regulated genes associated with neuronal and vascular development and differentiation, axonogenesis, glial cell differentiation, hypoxia-induced factor 1 pathway, and neuronal growth factor pathways. These changes were prevented by GSDMD-KO. GSDMD-KO alleviates hyperoxia-induced inflammatory injury, cell survival and death, and alterations of transcriptional gene expression of pathways involved in neuronal growth, development, and differentiation in the hippocampus of neonatal mice. This suggests that GSDMD plays a pathogenic role in preterm brain injury, and targeting GSDMD may be beneficial in preventing and treating brain injury and poor neurodevelopmental outcomes in preterm infants. The online version contains supplementary material available at 10.1186/s12974-023-02878-8.
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