scRNA-seq analysis of the livers from the endothelial cell-specific CTGF KO mice after pIVCL

We performed scRNA-seq analysis on the livers from the endothelial cell-specific CTGF knockout (KO) mouse or control wild-type (WT) littermate after partial inferior vena cava ligation (pIVCL), a mouse model of congestive hepatopathy. The scRNA-seq analysis showed that CTGF knockout in LSECs suppressed the expression of YAP/TAZ target genes and basement membrane-related genes in LSECs caused by hepatic congestion. Overall design: After liver perfusion with pronase and collagenase, hepatocytes were removed via low-speed centrifugation, and dead cells and erythrocytes were removed via density gradient centrifugation.

我们针对经部分下腔静脉结扎（partial inferior vena cava ligation, pIVCL）构建的充血性肝病小鼠模型，采集内皮细胞特异性结缔组织生长因子（connective tissue growth factor, CTGF）敲除（knockout, KO）小鼠及其野生型（wild-type, WT）同窝对照小鼠的肝脏组织，开展单细胞RNA测序（single-cell RNA sequencing, scRNA-seq）分析。测序结果显示，肝窦内皮细胞（liver sinusoidal endothelial cells, LSECs）中CTGF的敲除可抑制肝充血诱导的LSECs内YAP/TAZ（Yes-associated protein/transcriptional coactivator with PDZ-binding motif）靶基因及基底膜相关基因的表达。本研究的总体实验设计如下：采用链霉蛋白酶与胶原酶对肝脏进行灌注后，通过低速离心去除肝细胞，再经密度梯度离心清除死细胞与红细胞。