mRNA-sequencing uncovered the underlying mechanisms of xylazine related liver injury

Objective: The present study is aimed to study the liver injury in xylazine poisoning and uncover the underlying mechanism. Method: Forty male SD rats were randomly dived into four groups, control (saline), low dose (10mg/kg xylazine), median dose (20mg/kg xylazine) and high dose (40mg/kg xylazine). And the rats were injected the drug intraperitoneal continuous 28 days, and then collected serum and liver tissues, Elisa, RNA sequencing, histopathology examination, RT-qPCR were performed. Results: Compared to the control group, the body weight of 40mg/kg group was decreased, the level of ALT and AST in serum of 40mg/kg group was increased. Histopathological examination showed fatty degeneration, necrosis and fibrosis of the liver. 192 up-regulated and 277 down-regulated genes were identified through RNA sequencing in the 40mg/kg group and the PPAR signaling pathway ranked first in the KEGG pathway analysis, The common DEG in the 10mg/kg and 40mg/kg (Lox), four common DEGs in the 20mg/kg and 40mg/kg (Srebf1, Nr1dl, Fasn, SCD1), and four genes PCK1, FABP5, ACOX2, CPT2 in the PPAR signaling pathway in the 40mg/kg group were validated through Real-Time PCR Analysis. Conclusion：Long term xylazine injection can cause liver injury and the PAPR signaling pathway play a core role in the process of xylazine related liver injury.研究目的：本研究旨在探究赛拉嗪（xylazine）中毒所致的肝损伤，并揭示其潜在分子机制。 研究方法：将40只雄性SD大鼠随机分为四组，分别为对照组（给予生理盐水）、低剂量组（10mg/kg赛拉嗪）、中剂量组（20mg/kg赛拉嗪）与高剂量组（40mg/kg赛拉嗪）。采用腹腔注射方式连续给药28天，随后采集大鼠血清与肝组织样本，依次开展酶联免疫吸附测定（ELISA）、RNA测序、组织病理学检查以及实时定量聚合酶链反应（RT-qPCR）检测。 研究结果：与对照组相比，40mg/kg高剂量组大鼠体重显著降低，血清中谷丙转氨酶（ALT）与谷草转氨酶（AST）水平均升高。组织病理学检查显示，大鼠肝脏出现脂肪变性、坏死及纤维化病变。通过RNA测序分析，高剂量组共鉴定出192个上调差异表达基因与277个下调差异表达基因；KEGG通路富集分析显示，过氧化物酶体增殖物激活受体（PPAR）信号通路位列首位。经实时定量PCR验证，10mg/kg组与40mg/kg组共有的差异表达基因（Lox）、20mg/kg组与40mg/kg组共有的4个差异表达基因（Srebf1、Nr1dl、Fasn、SCD1），以及40mg/kg组PPAR信号通路上的4个关键基因（PCK1、FABP5、ACOX2、CPT2）的表达水平均存在显著异常。 研究结论：长期注射赛拉嗪可引发肝损伤，PPAR信号通路在赛拉嗪相关肝损伤过程中发挥核心调控作用。