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• W4381337488 abstract "Abstract Background & Aims: Liver sinusoidal endothelial cells (LSECs) constitute the wall of the hepatic sinusoids, which are the first to be affected by various injuries and play an important role in the occurrence and development of chronic liver diseases caused by different etiologies. Using a combination of bulk RNA sequencing (bulk RNA-seq) and single-cell RNA sequencing (scRNA-seq) techniques, we explored the characteristics of human primary LSECs during liver cirrhosis. Methods: A single-cell suspension containing human hepatic cells was prepared by circulating collagenase perfusion, and primary LSECs were isolated by density gradient centrifugation and magnetic-activated cell sorting. Immunofluorescence staining was used to identify the isolated LSECs, and cell purity was determined by flow cytometry. scRNA-seq analysis was performed on liver tissues from three patients with a non-cirrhotic liver and four patients with a cirrhotic liver. LSECs were annotated using gene expression patterns, and the differentially expressed genes (DEGs) were compared between the two groups. LSECs from seven patients with a non-cirrhotic liver and four patients with a cirrhotic liver were included in the bulk RNA-seq, and DEGs during liver cirrhosis were analyzed. The results of bulk RNA-seq and scRNA-seq were compared to screen for hub genes with similar trends. Quantitative real-time PCR, western blotting, and tissue immunofluorescence were performed on genes with significant differences to verify dysregulation of transcription and protein expression levels. Results: A total of 1987 DEGs were identified in bulk RNA-seq, and 461 DEGs were identified in scRNA-seq. A total of 103 common DEGs were identified by combining the two sets of sequencing data. Further screening was performed based on changes in expression levels, and 24 DEGs were retained for additional analysis. There were 19 up-regulated genes (FGF23, CH25, ANGPTL4, IL1RL1, CYP3A7, SERPINE1, F2RL3, MT1M, APOLD1, ADM, MFAP4, ERRFI1, CD9, LCN6, AKAP12, PLIN2, BNIP3, CAV2, and MT1A) and five down-regulated genes (GPR182, HSPA1B, RGS16, HSPA6, and MT-ND4L). Among them, FGF23 mRNA expression was the most up-regulated during cirrhosis. Correspondingly, FGF23 protein levels in LSECs from cirrhotic livers also increased. Conclusions: Through transcriptome analysis, DEGs related to the pathophysiological processes of LSECs during liver cirrhosis were identified, providing promising targets and pathways for the diagnosis and treatments of liver cirrhosis." @default.
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• W4381337488 date "2023-06-20" @default.
• W4381337488 modified "2023-10-16" @default.
• W4381337488 title "Transcriptome analysis of liver sinusoidal endothelial cells in patients with liver cirrhosis" @default.
• W4381337488 doi "https://doi.org/10.21203/rs.3.rs-3027266/v1" @default.
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