Transcriptomic Profiling of Myeloid Cells in Acetaminophen-Induced Acute Liver Failure. Transcriptomic Profiling of Myeloid Cells in Acetaminophen-Induced Acute Liver Failure

In the recent study, we found that mesenchymal stem cell (MSC)-derived extracellular vesicles expressing the SIRPα protein (SIRP-EVs) were significantly distributed within myeloid cells (CD11b+ cells) and kupffer cells (CD11b+/F4/80+ cells) in acetaminophen (APAP)-induced mouse acute liver failure (ALF) model. Furthermore, SIRP-EVs enhanced the phagocytic activity of macrophages by blocking CD47 on necroptotic hepatocytes and promoted liver regeneration. Therefore, we investigated the therapeutic effects of SIRP-EVs on CD11b+ cells in APAP-induced ALF conditions. CD11b+ cells in the liver, including resident and recruited CD11b+ cells, were harvested and analyzed through bulk RNA sequencing. Overall design: APAP was administered intraperitoneally to induce ALF, and EVs were administered intravenously 8 hours after APAP induction. 48 hours after APAP induction, livers were harvested and dissociated into single cells. CD11b+ cells were then immediately isolated from liver single-cell suspensions and analyzed via bulk RNA-sequenceing.

本项近期研究表明，表达SIRPα蛋白（SIRPα protein）的间充质干细胞（mesenchymal stem cell, MSC）来源细胞外囊泡（extracellular vesicles, EVs，命名为SIRP-EVs），在对乙酰氨基酚（acetaminophen, APAP）诱导的小鼠急性肝衰竭（acute liver failure, ALF）模型中，显著分布于髓系细胞（CD11b阳性细胞，CD11b+ cells）以及库普弗细胞（CD11b+/F4/80阳性细胞，CD11b+/F4/80+ cells）内。进一步研究证实，SIRP-EVs可通过阻断坏死性凋亡肝细胞表面的CD47分子，增强巨噬细胞的吞噬活性，并促进肝脏再生。据此，本研究探讨了SIRP-EVs在对乙酰氨基酚诱导的急性肝衰竭模型中对CD11b阳性细胞的治疗作用。
本研究收集肝脏内的CD11b阳性细胞（包括定居性及招募性CD11b阳性细胞），通过批量RNA测序进行分析。实验整体设计如下：通过腹腔内注射对乙酰氨基酚构建小鼠急性肝衰竭模型，于造模后8小时经静脉输注SIRP-EVs；造模后48小时处死小鼠并获取肝脏组织，将其解离为单细胞悬液，随后立即从肝脏单细胞悬液中分离CD11b阳性细胞，通过批量RNA测序完成检测分析。