NHC Key Laboratory of Combined Multi-organ Transplantation, Hangzhou, 310003, Zhejiang, China.
Zhejiang University School of Medicine, Hangzhou, 310058, Zhejiang, China.
Mol Biomed. 2024 Jun 21;5(1):22. doi: 10.1186/s43556-024-00185-z.
Hepatic ischemia-reperfusion injury (HIRI) is a critical pathophysiological process during liver transplantation (LT). Multiple genes and signal pathways are dysregulated during HIRI. This study aims to identify genes as potential therapeutic targets for ameliorating HIRI. Datasets containing samples from the human donor liver (GSE151648) and mouse HIRI model (GSE117066) were analyzed to determine differentially expressed genes (DEGs). The selected DEGs were confirmed by real-time PCR and western blot in the hepatocyte hypoxia-reoxygenation (HR) model, mouse HIRI model, and human liver samples after transplantation. Genetic inhibition was used to further clarify the underlying mechanism of the gene in vitro and in vivo. Among the DEGs, CSRNP1 was significantly upregulated (|log FC|= 2.08, P < 0.001), and was positively correlated with the MAPK signal pathway (R = 0.67, P < 0.001). CSRNP1 inhibition by siRNA significantly suppressed apoptosis in the AML-12 cell line after HR (mean Annexin ratio = 60.62% vs 42.47%, P = 0.0019), but the protective effect was eliminated with an additional MAPK activator. Knocking down CSRNP1 gene expression by intravenous injection of AAV-shRNA markedly reduced liver injury in mouse HIRI model (ALT: AAV-NC vs AAV-shCsrnp1 = 26,673.5 ± 2761.2 vs 3839.7 ± 1432.8, P < 0.001; AST: AAV-NC vs AAV-shCsrnp1 = 8640.5 ± 1450.3 vs 1786.8 ± 518.3, P < 0.001). Liver-targeted delivery of siRNA by nanoparticles effectively inhibited intra-hepatic genetic expression of Csrnp1 and alleviated IRI by reducing tissue inflammation and hepatocyte apoptosis. Furthermore, CSRNP1 inhibition was associated with reduced activation of the MAPK pathway both in vitro and in vivo. In conclusion, our results demonstrated that CSRNP1 could be a potential therapeutic target to ameliorate HIRI in an MAPK-dependent manner.
肝缺血再灌注损伤(HIRI)是肝移植(LT)过程中的一个关键病理生理过程。在 HIRI 过程中,多个基因和信号通路发生失调。本研究旨在鉴定基因作为改善 HIRI 的潜在治疗靶点。分析了包含人类供体肝样本(GSE151648)和小鼠 HIRI 模型样本(GSE117066)的数据集,以确定差异表达基因(DEGs)。通过实时 PCR 和蛋白质印迹在肝细胞缺氧-复氧(HR)模型、小鼠 HIRI 模型和移植后人类肝样本中验证了所选 DEGs。遗传抑制用于进一步阐明该基因在体外和体内的潜在机制。在 DEGs 中,CSRNP1 显著上调(|log FC|=2.08,P<0.001),并与 MAPK 信号通路呈正相关(R=0.67,P<0.001)。siRNA 抑制 CSRNP1 可显著抑制 HR 后 AML-12 细胞系的凋亡(平均 Annexin 比值=60.62% vs 42.47%,P=0.0019),但用额外的 MAPK 激活剂则消除了保护作用。通过静脉注射 AAV-shRNA 敲低 CSRNP1 基因表达可显著减轻小鼠 HIRI 模型中的肝损伤(ALT:AAV-NC 与 AAV-shCsrnp1=26673.5±2761.2 与 3839.7±1432.8,P<0.001;AST:AAV-NC 与 AAV-shCsrnp1=8640.5±1450.3 与 1786.8±518.3,P<0.001)。通过纳米颗粒进行肝靶向 siRNA 递送可有效抑制 Csrnp1 的肝内遗传表达,并通过减少组织炎症和肝细胞凋亡来减轻 IRI。此外,CSRNP1 抑制与 MAPK 通路的体外和体内激活减少有关。总之,我们的研究结果表明,CSRNP1 可能是一种潜在的治疗靶点,可通过 MAPK 依赖性方式改善 HIRI。