LPS诱导脓毒症小鼠脾组织microRNA表达的筛选研究

目的采用Small RNA测序技术检测脓毒症小鼠24 h脾组织microRNA(miRNA)的表达变化,并从非编码基因层面分析脓毒症脾功能损伤的可能机制。方法将30只SPF级健康BALB/C小鼠随机分为对照组和脓毒症组,每组15只。脓毒症组采用LPS 10 mg/kg腹腔注射进行建模,对照组仅予等量生理盐水腹腔注射。两组术毕均行肌肉注射平衡液5 ml/kg,并于术后24 h各组随机取5只小鼠的脾组织提取RNA,采用Small RNA测序技术进行miRNA检测,然后应用生物信息学软件分析脓毒症小鼠脾组织的miRNA表达差异及靶基因预测。结果小鼠建模后24 h,与对照组相比,脓毒症小鼠脾脏组织已知及未知miRNA表达上调数分别为39个、3个,下调数分别为16个、3个。其中已知miRNA表达上、下调倍数最大的前5个基因分别为Rno-miR-217-5p,rno-mir-216a-5p,rnomiR-216b-5p,rno-miR-375-3p,rno-mir-216b-3p。下调倍数前5的miRNA:Rno-miR-138-5p,rno-miR-451a-3p,rno-miR-138-1-3p,rno-miR-202-5p,rno-miR-592。结论LPS诱导脓毒症小鼠可出现miRNA表达量的改变,其上、下调倍数最显著的miRNA可能对预测LPS诱导的脓毒症脾功能损伤有一定指导意义。

Expression of microRNAs in spleen of septic mice induced by LPS

Objective To detect the expression of microRNAs in 24 h spleen tissue of septic mice by using Small RNA sequencing technology,and then to analyze the possible mechanism of spleen injury induced by LPS from the level of non-coding genes.Methods Thirty SPF grade healthy BALB/C mice were randomly divided into control group and sepsis group.The mice of sepsis group were modeled by intraperitoneal injection of LPS 10 mg/kg.The control group was given intraperitoneal injection of the same amount of normal saline.The two groups received intramuscular injection of 5 ml/kg balance fluid after modeling.At 24 h after surgery,5 mice were randomly selected from each group to extract RNA from spleen tissues and then used Small RNA sequencing technology for microRNA detection.Bioinformatics software was used to analyze microRNA expression differences and target gene prediction in spleen tissues of sepsis mice.Results 24 hours after modeling,the number of up-regulation of known and unknown microRNA expression in spleen tissues of sepsis mice was 39 and 3,and the down-regulation was 16 and 3,respectively,relative to the control group.Among them,the top 5 genes with the highest microRNA expression up-regulation are rno-miR-216b-5p,rno-miR-375-3p,rno-miR-216b-3p,rno-miR-217-5p,rno-miR-216a-5p;rno-miR-138-1-3p,rno-miR-202-5p,rno-miR-592,rno-miR-138-5p,rno-miR-451-3p.The corresponding target gene prediction is detailed in the results section.Conclusion The miRNA expression in LPS induced sepsis mice can be changed.The miRNA with the most significant up and down-regulation may be of guiding significance to predict the spleen function damage induced by LPS.