MicroRNA-674-5p/5-LO axis is important in concanavalin A-induced acute mouse liver injury. Mus musculus

Eight-week-old male pathogen-free BALB/c mice were used for the induction of concanavalin A (Con A)-induced hepatitis. Animals were acclimated for at least 6-7 days to a 12-hour light/dark cycle in a humidity- and temperature-controlled, specific-pathogen-free environment in autoclaved cages. Mice were randomly assigned to two groups (4 in each group) and received Con A type IV (or saline) intravenously via the tail vein at the dose of 20 mg/kg for 2 hours. The animals were sacrificed for blood and liver collection. Blood was taken for transaminase and cytokine measurements. Livers were excised immediately after sacrifice. Part of the livers were immediately snap-frozen in liquid nitrogen and kept at -80°C for total RNA isolation and microRNA microarray analysis. miR-674-5p and other miRNAs were experimentally validated to be dysregulated by Con A in mouse livers. Overall design: Liver samples from four mice treated with Con A and four mice treated with saline were used to acquire the miRNA expression profiling.

本研究采用8周龄雄性无特定病原体（pathogen-free, SPF）BALB/c小鼠，构建刀豆蛋白A（concanavalin A，Con A）诱导的肝炎模型。实验动物于高压灭菌笼具中，在温湿度可控的无特定病原体环境内，以12小时明暗循环条件适应饲养至少6~7天。将小鼠随机分为两组（每组4只），分别经尾静脉注射IV型刀豆蛋白A（Con A）或生理盐水，给药剂量为20 mg/kg，处理2小时后处死动物，采集血液与肝脏样本。采集的血液用于转氨酶与细胞因子水平检测；处死后立即摘取肝脏，取部分肝脏样本即刻置于液氮中快速冷冻，并保存于-80℃环境，用于总RNA提取及microRNA（miRNA）芯片分析。经实验验证，小鼠肝脏中miR-674-5p及其他microRNA的表达水平经Con A处理后发生失调。实验整体设计：本研究选取4只Con A处理组小鼠与4只生理盐水对照组小鼠的肝脏样本，开展microRNA表达谱分析。