【摘要】 目的应用RNA-seq技术检测脓毒症大鼠24、48 h肝组织基因的表达变化,分析脓毒性肝损伤可能的机制。方法随机将30只Wistar大鼠等分为脓毒症组和对照组。脓毒症组采用盲肠结扎穿孔术(CLP)建模,对照组仅行开腹、关腹。两组术毕均肌肉注射平衡液5 mL/kg。术后24、48 h各组随机取5只大鼠取肝组织提取总RNA后采用RNA-seq进行mRNA检测,应用生物信息学软件分析脓毒症大鼠肝组织mRNA随时间变化的表达差异及涉及的相关通路。结果大鼠建模后24、48 h,相对于对照组,脓毒症大鼠肝脏组织mRNA表达上调数分别为1220个、914个,下调数分别为1741个、822个。其中脓毒症大鼠肝组织随时间变化,NF-κB、信号通路、PPAR信号通路相关mRNA表达差异明显。脓毒症大鼠24 h肝组织NF-κB信号通路相关基因正常表达,而48 h转为上调有13个,下调转正常表达有4个,下调转上调有1个,上调转下调有1个,上调转为正常表达有4个,正常表达转为下调有6个,同时上调有7个,同时下调有1个。PPAR信号通路相关的基因从下调转为正常表达有20个,上调转正常表达有3个,正常表达转上调有4个,同时下调有6个,同时上调有5个。结论肝脏炎症反应、代谢改变可能是脓毒性肝损伤的重要机制之一,可能跟NF-κB、P信号通路、PAR信号通路相关基因表达有关。更多还原

【Abstract】 Objective To detect the changes of mRNA expression of liver tissue in sepsis rats at24 h and 48 h by RNA-seq technique,and to analyze the possible mechanism of liver injury in sepsis.Methods 30 healthy male Wistar rats of clean grade were divided into control group and sepsis group by random number table.Cecal ligation and perforation(CLP) was used to make sepsis models.The control group only underwent laparotomy and abdominal closure.The balance fluid was injected intramuscularly at 5 mL/kg in two groups.Each group,the livers of 5 rats picked at random were harvested at 24 h and 48 h after modeling.RNA-seq was used to detect the expression of mRNA in liver tissues of sepsis rats at 24 h and 48 h after operation.The time-varying expression of mRNA in liver tissues of sepsis rats and the related pathways were analyzed by bioinformatics software.Results At 24 h and 48 h after modeling,compared with the control group,the up-regulation of liver tissue mRNA expression in sepsis rats were 1220,914 respectively and down-regulation were 1741,822 respectively.Among of them,the expression of NF-κB signaling pathway,PPAR signaling pathwayrelated genes in the livers of sepsis rats varied significantly with time.The expression of NF-κB signaling pathway related genes in liver tissue of sepsis rats was normal at 24 h,13 genes up-regulated at 48 h,4 genes down-regulated to normal expression,1 gene was from down-regulation to upregulation,1 gene from up-regulation to down-regulation,4 genes from up-regulation to normal expression,6 genes from normal expression to down-regulation,7 genes up-regulated at two points at the same time and 1 gene down-regulated at two points.There were 20 genes related to PPAR signaling pathway from down-regulation to normal expression,3 from up-regulation to normal expression,4 from normal expression to up-regulation,6 genes down-regulated at two points concurrently and 5 genes up-regulated at two points.Conclusion The changes of liver inflammation and metabolism may be one of the important mechanisms of liver injury in sepsis.The mechanism may be related to the changes of gene expression related to NF-κB signaling pathway and PPAR signaling pathway.更多还原