Liu Yong, Jiang Xin, Zhang Yunfei, Tong Guofeng, Tang Kai, Gui Yanlin, Wen Lan, Li Changqing
School of Clinical Medicine, Chengdu Medical College, Chengdu, Sichuan 610500, China; Department of Neurology, The First Affiliated Hospital of Chengdu Medical College, Chengdu, Sichuan 610500, China.
School of Pharmacy Chengdu Medical College, Chengdu, Sichuan 610500, China.
Exp Neurol. 2025 Mar;385:115127. doi: 10.1016/j.expneurol.2024.115127. Epub 2024 Dec 22.
MicroRNAs (miRNAs) are widely involved in signal transduction and regulation during cerebral ischemia-reperfusion injury (CIRI). This study investigates the molecular mechanisms of the specific miRNA/DDX3X/NLRP3 pathway in early-stage CIRI and explores its potential clinical applications. Through public database analysis, miR-135a-5p targeting DDX3X after CIRI was determined. The levels of DDX3X, NLRP3 inflammasome, and GSDMD-N were increased after MCAO/R. Upregulation of miR-135a-5p suppressed these protein levels. Upregulating miR-135a-5p also reduced infarct volume and neuronal pyroptosis, while improved neurological scores in MCAO/R mice. Co-IP confirmed protein interaction between DDX3X and NLRP3 in CIRI models. Furthermore, miR-135a-5p mimics alleviated pyroptosis and inhibited DDX3X/NLRP3 pathway activation after OGD/R cells, whereas miR-135a-5p inhibitor produced the opposite effect. The dual-luciferase reporter assay validated that DDX3X was a direct target of miR-135a-5p. Clinically, the serum level of miR-135a-5p was significantly lower in CIRI patients after thrombectomy compared to controls. The levels of DDX3X, NLRP3, and IL-18 were elevated in the CIRI group, while the difference of IL-1β levels between the two groups was not statistically significant (p = 0.055). Although an inverse correlation was observed between miR-135a-5p and DDX3X levels in CIRI patients, the linear regression analysis did not reach statistical significance (R = 0.12, p = 0.061). This study indicated that miR-135a-5p/DDX3X/NLRP3 pathway is pivotal in early-stage CIRI. Upregulation of miR-135a-5p inhibits NLRP3-mediated neuronal pyroptosis by targeting DDX3X, thereby alleviating CIRI and improving neurological function. This signaling axis holds promise for future clinical applications in treating CIRI.
微小RNA（miRNA）广泛参与脑缺血再灌注损伤（CIRI）过程中的信号转导和调节。本研究探讨了特定miRNA/DDX3X/NLRP3通路在早期CIRI中的分子机制，并探索其潜在的临床应用。通过公共数据库分析，确定了CIRI后靶向DDX3X的miR-135a-5p。大脑中动脉闭塞/再灌注（MCAO/R）后，DDX3X、NLRP3炎性小体和GSDMD-N的水平升高。miR-135a-5p的上调抑制了这些蛋白质水平。上调miR-135a-5p还减少了梗死体积和神经元焦亡，同时改善了MCAO/R小鼠的神经功能评分。免疫共沉淀（Co-IP）证实了CIRI模型中DDX3X与NLRP3之间的蛋白质相互作用。此外，miR-135a-5p模拟物减轻了氧糖剥夺/再灌注（OGD/R）细胞后的焦亡，并抑制了DDX3X/NLRP3通路的激活，而miR-135a-5p抑制剂则产生相反的效果。双荧光素酶报告基因检测验证了DDX3X是miR-135a-5p的直接靶点。临床上，与对照组相比，血栓切除术后CIRI患者血清miR-135a-5p水平显著降低。CIRI组中DDX3X、NLRP3和IL-18水平升高，而两组间IL-1β水平的差异无统计学意义（p = 0.055）。虽然在CIRI患者中观察到miR-135a-5p与DDX3X水平呈负相关，但线性回归分析未达到统计学意义（R = 0.12，p = 0.061）。本研究表明，miR-135a-5p/DDX3X/NLRP3通路在早期CIRI中起关键作用。miR-135a-5p的上调通过靶向DDX3X抑制NLRP3介导的神经元焦亡，从而减轻CIRI并改善神经功能。该信号轴在治疗CIRI的未来临床应用中具有前景。
