Wang S, Ruan Q F, Xie W G, Chen L, Jiang M J, Ruan J J, Ye Z Q
Department of General Surgery, Zhongnan Hospital of Wuhan University, Wuhan 430071, China.
Zhonghua Shao Shang Za Zhi. 2017 Oct 20;33(10):639-643. doi: 10.3760/cma.j.issn.1009-2587.2017.10.012.
To explore differential expression of microRNAs in serum of patients with severe burn and analysis of the signaling pathway at early stage. In this study, we included three healthy adult volunteers and three patients with severe burn, conforming to the inclusion criteria and hospitalized in Tongren Hospital of Wuhan University & Wuhan Third Hospital in July 2015. Venous whole blood of 6 mL of each burn patient and healthy volunteer was collected at 24 to 48 h post injury of burn patients. The whole blood was divided into burn group and healthy control group. Whole blood of 2 mL of each one was used to determine white blood cell count and neutrophile granulocyte content. Serum was separated from the other whole blood of 4 mL of each one. Half of serum was used to determine content of blood glucose, total protein, and albumin; another half of serum was used to extract total RNA with Trizol method. The differentially expressed microRNA, with differential expression ratio larger than or equal to 1.500 between 2 groups, were screened by microRNA chip technique. Then cluster analysis and functional enrichment analysis of Kyoto encyclopedia of genes and genomes (KEGG) signaling pathway were performed on the differentially expressed microRNAs. Data were processed with test. (1) Content of white blood cell count, neutrophile granulocyte of whole blood, and blood glucose of serum of patients in burn group was obviously higher than that in healthy control group (with values from 4.27 to 7.83, <0.05 or <0.01). Content of total protein and albumin of serum of patients in burn group was significantly lower than that in healthy control group (with values respectively -12.80 and -12.36, values below 0.01). (2)Compared with those in serum of healthy control group, differential expression ratios of 48 microRNAs in serum of burn group were larger than 1.500, with 22 up-regulated microRNAs and 26 down-regulated microRNAs. MicroRNA expression profile in serum of burn group was different from that of healthy control group. (3)Functional enrichment analysis of KEGG signaling pathway showed that compared with those in serum of healthy control group, microRNAs of differential expression in serum of burn group took part in tumor transcription misregulation signaling pathway, tumor proteoglycan signaling pathway, long-term potentiation signaling pathway, tumor associated microRNAs signaling pathway, citrate cycle signaling pathway, tumor necrosis factor signaling pathway, focal adhesion signaling pathway, endocytosis signaling pathway, insulin secretion signaling pathway, and estrogen signaling pathway. MicroRNA expression profile in serum of patients with severe burn is different from that in serum of healthy adults. MicroRNAs of differential expression may take part in important pathophysiological process of energy metabolism, inflammatory response, and regulation of blood glucose at early stage of severe burn.
探讨严重烧伤患者血清中微小RNA的差异表达及早期信号通路分析。本研究纳入3名健康成年志愿者和3例严重烧伤患者,均符合纳入标准,于2015年7月在武汉大学同仁医院及武汉市第三医院住院治疗。在烧伤患者受伤后24至48小时采集每名烧伤患者和健康志愿者6 mL静脉全血。全血分为烧伤组和健康对照组。每组取2 mL全血用于测定白细胞计数和中性粒细胞含量。从每组另外4 mL全血中分离血清。一半血清用于测定血糖、总蛋白和白蛋白含量;另一半血清用Trizol法提取总RNA。采用微小RNA芯片技术筛选两组间差异表达倍数大于或等于1.500的差异表达微小RNA。然后对差异表达的微小RNA进行基因与基因组京都百科全书(KEGG)信号通路的聚类分析和功能富集分析。数据采用检验进行处理。(1)烧伤组患者全血白细胞计数、中性粒细胞含量及血清血糖含量均明显高于健康对照组(值为4.27至7.83,<0.05或<0.01)。烧伤组患者血清总蛋白和白蛋白含量明显低于健康对照组(值分别为-12.80和-12.36,值均低于0.01)。(2)与健康对照组血清相比,烧伤组血清中48种微小RNA的差异表达倍数大于1.500,其中22种微小RNA上调,26种微小RNA下调。烧伤组血清微小RNA表达谱与健康对照组不同。(3)KEGG信号通路功能富集分析显示,与健康对照组血清相比,烧伤组血清中差异表达的微小RNA参与肿瘤转录失调信号通路、肿瘤蛋白聚糖信号通路、长时程增强信号通路、肿瘤相关微小RNA信号通路、柠檬酸循环信号通路、肿瘤坏死因子信号通路、粘着斑信号通路、内吞作用信号通路、胰岛素分泌信号通路和雌激素信号通路。严重烧伤患者血清微小RNA表达谱与健康成年人血清不同。差异表达的微小RNA可能参与严重烧伤早期能量代谢、炎症反应和血糖调节等重要病理生理过程。