摘要
目的:探索敌草快中毒时肾小管上皮细胞内显著变化的基因和信号通路,为阐明敌草快中毒相关肾损伤的病理机制和优化临床治疗措施提供科学依据。方法:通过灌胃给药构建小鼠敌草快中毒模型,分析染毒后肾脏形态学和功能学的变化,并分选小鼠肾小管上皮细胞进行mRNA和microRNA测序。筛选敌草快中毒后显著差异的基因并进行功能富集,分析中毒组肾小管上皮细胞内显著激活的信号通路。结果:(1)敌草快中毒后小鼠血浆中IL-6、肌酐和尿素氮含量显著升高;肾脏肿胀,暗红质脆;肾小管上皮细胞肿胀,部分空泡变性,偶见坏死;红细胞管型和蛋白管型致管腔狭窄甚至闭塞;肾间质充血伴炎症细胞浸润。(2)中毒后肾脏上皮细胞1925种基因上调,3858种基因下调,其中促炎因子(TNF和MMP8)、趋化因子(CCL2、CXCL1和CXCL2)和促凋亡分子(BAK、Tnfsf14和Caspase3)显著增加,抑炎因子(SOCS和TGF-β)以及抑制凋亡分子(BCL2)显著降低。(3)中毒后144种microRNA上调,146种microRNA降低,其中microRNA-21可靶向调控差异基因MMP-8和VCAM-1;microRNA-30a与BCL2L11存在靶向关系。(4)GO、KEGG和GSEA功能分析提示差异基因在程序性细胞死亡、氧化应激和炎症反应相关信号通路中富集,主要包括凋亡、IL-17、NF-κB和MAPK等信号通路。结论:敌草快中毒后肾脏显著损伤,肾小管上皮细胞内促炎因子和促凋亡分子增加,抑炎和抑凋亡分子降低;程序性细胞死亡、氧化应激和炎症反应等信号通路参与敌草快中毒相关肾损伤。未来,可针对本研究中的核心分子开发靶向治疗药物,优化临床策略,改善患者预后。
Objective:To explore the genes and signal pathways that significantly change in renal tubular epithelial cells of Diquat-poisoned mice,in order to provide scientific basis for clarifying the pathological mechanism of diquat related renal injury and optimizing clinical treatment measures.Methods:We established a model of Diquat-poisoned mice by intragastric administration.The changes of renal morphology and function were analyzed after exposured to Diquat,and the renal tubular epithelial cells of mice were sorted for mRNA and microRNA sequencing.After screening and functional enrichment of genes with significant differences after Diquat poisoning,We analyzed the signal pathways which significantly activated in renal tubular epithelial cells in poisoning groups.Results:(1)The plasma concentrations of IL-6,creatinine and urea nitrogen of Diquatpoisoned mice were significantly increased.After exposured to Diquat,the kidney swelled,dark red and fragile.The epithelial cells of renal tubules swelled,some of which showed vacuolar degeneration or necrosis.The lumen of the renal tubules is blocked by red blood cell and protein tubular types.Many inflammatory cells infiltrated the renal interstitium.(2)1925 kinds of genes were up-regulated and 3858 genes were down-regulated in renal epithelial cells after exposured to Diquat.The proinflammatory factors(TNF and MMP8),chemokines(CCL2,CXCL1 and CXCL2)and pro-apoptotic molecules(BAK,Tnfsf14 and Caspase3)were significantly increased.Anti-inflammatory factors(SOCS and TGF-β)and anti-apoptotic molecule(BCL2)were significantly reduced.(3)144 kinds of microRNAs were up-regulated and 146 microRNAs were down-regulated in poisoned renal epithelial cells.Significantly increased microRNA-21 may target MMP-8 and VCAM-1.There was also a targeting relationship between microRNA-30a and BCL2L11.(4)The differentially expressed genes were enriched in signal pathways related to programmed cell death,oxidative stress and inflammatory response,mainly including apoptosis,IL-17,NF-κB and MAPK sign
作者
李琳
刘善收
虎晓岷
王仙琦
代铮
梁晓丽
LI Lin;LIU Shanshou;HU Xiaomin;WANG Xianqi;DAI Zheng;LIANG Xiaoli(Emergency Department of Qin Huang Hospital,Xi'an 710699,China;Emergency Department of the First Affiliated Hospital of PLA Air Force Medical University,Xi'an 710032,China)
出处
《现代医学》
2023年第4期432-441,共10页
Modern Medical Journal
基金
陕西省自然科学基础研究项目(2020JQ-466)。
关键词
敌草快中毒
多组学测序
肾小管上皮细胞
病理机制
靶向治疗
小鼠
Diquat poisoning
multigroup sequencing
renal tubular epithelial cells
pathological mechanisms
targeted therapy
mice
