Department of Neurosurgery, Zhongshan Hospital, Fudan University, Shanghai, China.
National Clinical Research Center for Interventional Medicine of China, Shanghai, China.
DNA Cell Biol. 2023 Sep;42(9):554-562. doi: 10.1089/dna.2023.0106. Epub 2023 Aug 11.
Inhibition of the inflammatory response triggered by microglial pyroptosis inflammatory activation may be one of the effective ways to alleviate cerebral ischemia-reperfusion injury, the specific mechanism of which remains unclear. In this study, BV-2 microglia with or without oxygen-glucose deprivation/reoxygenation (OGD/R) or long noncoding RNA (lncRNA) Gm44206 knockdown were used as cell models to conduct an study. Detection of lactate dehydrogenase release and pyroptosis-related protein levels was performed using a corresponding kit and western blotting, respectively. Proliferation of microglia was evaluated by CCK8 assay. Enzyme-linked immunosorbent assay was applied for measuring levels of proinflammatory cytokines. This study verified the involvement of microglial pyroptosis as well as upregulation of NLRP3, Caspase-1, GSDMD, and Apoptosis-associated Speck-like protein containing a C-terminal caspase-recruitment domain (ASC) in cerebral ischemia-reperfusion injury. Moreover, knockdown of lncRNA Gm44206 could alleviate OGD/R-induced microglial pyroptosis and cell proliferation inhibition through the NLRP3/Caspase-1/GSDMD pathway, thus decreasing the release of proinflammatory cytokines, including interleukin (IL)-1β, IL-6, IL-18, and tumor necrosis factor-alpha. In conclusion, this study established a correlation between microglial pyroptosis and cerebral ischemia-reperfusion injury and identified lncRNA Gm44206 as a potential regulator of NLRP3/Caspase-1/GSDMD axis-mediated microglial pyroptosis, which could be considered a promising therapeutic target.
抑制小胶质细胞焦亡炎症激活引发的炎症反应可能是减轻脑缺血再灌注损伤的有效方法之一，但具体机制尚不清楚。本研究以氧葡萄糖剥夺/复氧（OGD/R）或长链非编码 RNA（lncRNA）Gm44206 敲低的 BV-2 小胶质细胞作为细胞模型进行研究。采用相应试剂盒和 Western blot 分别检测乳酸脱氢酶释放和焦亡相关蛋白水平。CCK8 法评估小胶质细胞增殖。酶联免疫吸附试验（ELISA）用于测定促炎细胞因子水平。本研究证实了小胶质细胞焦亡以及 NOD 样受体热蛋白结构域相关蛋白 3（NLRP3）、半胱氨酸天冬氨酸蛋白酶-1（Caspase-1）、Gasdermin D（GSDMD）和含有 C 端半胱氨酸天冬氨酸蛋白酶募集结构域（ASC）的凋亡相关斑点样蛋白（ASC）在脑缺血再灌注损伤中的上调。此外，lncRNA Gm44206 的敲低可通过 NLRP3/Caspase-1/GSDMD 通路减轻 OGD/R 诱导的小胶质细胞焦亡和细胞增殖抑制，从而减少包括白细胞介素（IL）-1β、IL-6、IL-18 和肿瘤坏死因子-α在内的促炎细胞因子的释放。综上所述，本研究建立了小胶质细胞焦亡与脑缺血再灌注损伤之间的相关性，并确定 lncRNA Gm44206 是 NLRP3/Caspase-1/GSDMD 轴介导的小胶质细胞焦亡的潜在调节因子，可作为有前途的治疗靶点。
