Inverse regulation of GSDMD and GSDME gene expression during LPS-induced pyroptosis in RAW264.7 macrophage cells
Inverse regulation of GSDMD and GSDME gene expression during LPS-induced pyroptosis in RAW264.7 macrophage cells
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DOI：
10.1007/s10495-022-01708-1
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发表时间：
2022-01-10
期刊：
影响因子：
7.2
通讯作者：
Uemura, Koichi
中科院分区：
文献类型：
--
作者：
Aki, Toshihiko;Funakoshi, Takeshi;Uemura, Koichi
GSDMD和GSDME是gasdermin蛋白家族的成员，参与质膜通道的形成，在称为焦亡的某种类型的坏死期间促成细胞破裂。GSDMD响应于免疫刺激如脂多糖（LPS）处理而被激活，而GSDME主要参与药物诱导的肿瘤细胞死亡。在此，我们发现，在LPS诱导的RAW264.7小鼠巨噬细胞焦亡过程中，GSDMD基因的表达显著增加。相反，在相同的细胞中GSDME表达降低。只有当细胞在高葡萄糖（4.5 g/l）培养基中培养时，才观察到LPS对GSDMD表达的增加作用，这表明葡萄糖的可用性对于这种作用是重要的。LPS对GSDMD表达的影响被2-脱氧葡萄糖（2DG）消除，证实糖酵解在LPS的增强作用中起关键作用。小干扰RNA介导的GSDMD敲低或GSDME过表达导致LPS诱导的焦亡通过GSDME而不是通过GSDMD发生。总而言之，LPS至少部分通过其对糖酵解的影响以相反的方向调节GSDMD和GSDME表达。这种转录调控可能有助于以GSDMD依赖的方式执行焦亡。
GSDMD and GSDME, members of the gasdermin protein family, are involved in the formation of plasma membrane channels contributing to cell rupture during a certain type of necrosis called pyroptosis. GSDMD is activated in response to immunological stimulation such as lipopolysaccharides (LPS) treatment while GSDME is mainly involved in drug-induced tumor cell death. Here we show that the expression of the GSDMD gene increases significantly during LPS -induced pyroptosis in RAW264.7 murine macrophage cells. In contrast, GSDME expression is decreased in the same cells. The increasing effect of LPS on GSDMD expression was observed only when the cells were cultured in high glucose (4.5 g/l) medium, suggesting that glucose availability is important for this effect. The effect of LPS on GSDMD expression is abolished by 2-deoxyglucose (2DG), confirming that glycolysis plays crucial roles in the increasing effect of LPS. Small interference RNA-mediated knock down of GSDMD or overexpression of GSDME causes LPS-induced pyroptosis to take place through GSDME rather than through GSDMD. Taken together, LPS regulates GSDMD and GSDME expression in opposite directions through, at least in part, its effect on glycolysis. This transcriptional regulation may contribute to the execution of pyroptosis in a GSDMD-dependent manner.