| Objective:Early diagnosis and effective treatment is the key factor for curing the acute lung injury patients with sepsis (ALI with sepsis).However, less is known about the related mechanism and molecular markers about ALI with sepsis. This study aimed to investigate the expression of miRNA, TLR4 and inflammatory cytokines (IL-34, IL-36) in plasma in acute lung injury patients with sepsis and its diagnosis significance in clinical application.Methods:Total of 18 patients with sepsis in Inner Mongolia Baotou hospital ICUC with 11 healthy controls) were enrolled in this study. Total RNA was extracted from peripheral blood by Trizol method, and the concentration and purity was detected by Nanodrop1000 ultraviolet spectrophotometer instrument. The total RNA samples of 6 cases of ALI patients with sepsis and 3 cases of healthy control were selected to detect miRNA expression with Agilent chip. Hybrid image is analyzed with Agilent Feature Extraction software, and the different expressed miRNA between ALI with sepsis and normal control group were screened by GeneSpring software. P values< 0.05 and FC (abs) value> 2 were considered significant difference. The function of the target genes related to different expressed miRNAs was analyzed with DAVID tools. The peripheral venous blood of 15 cases of ALI patients with sepsis and 5 cases of healthy control were was collected in 2% EDTA anticoagulant tube, and the supernatant after centrifugation was recollected to detect TLR4, IL-34, IL-36 by ELISA.Results:1. Total of 320 miRNAs with different expression profile are found in ALI patients with sepsis compared to healthy control, in which 138 miRNAs were up regulated and 182 miRNAs were down regulated. Among them, there are 7 miRNAs were up-regulated more than 10 times:miRNA-619-5p, miRNA-6881-3p, miRNA-449b-3p, miRNA-3657, miRNA-4254, miRNA-34b-3p and miRNA-1287-5p; There are 3 miRNAs were down-regulated more than 10 times: miRNA-362-5p, miRNA-892-b and miRNA-625-5p. These miRNAs were related to large number of predicted target genes. About 3500 of mutual target genes were screened by Venn Web tool. Results of target gene function analysis show that these mutual target genes mainly related to the regulation of biological membrane ion channels.2. TLR4 was significantly increased in ALI with sepsis. Levels of IL-34, IL-36 in peripheral blood were also significantly increased (P<0.01).3.The Pearson correlation analysis show that IL-34 concentration are moderately related to TLR4 concentration which are uncorrelated to IL-36 concentration.However, the concentration of IL-34 and IL-36 were positively correlated.Conclusion:320 different expressed miRNAs are found in ALI patients with sepsis compared to healthy control, in which 138 miRNAs were up regulated and 182 miRNAs were down regulated. About 3500 of mutual target genes of these miRNAs were mainly related to the regulation of biological membrane ion channels. TLR4, IL-34 and IL-36 levels in peripheral blood were significantly increased in ALI with sepsis. IL-34 concentration are positively related to TLR4 concentration.IL-34 have an effect on TLR4 and the expression of downstream inflammation factors in ALI with sepsis. IL-36 concentration are uncorrelated to TLR4 concentration. The concentration of IL-34 and IL-36 were correlated. IL-34, IL-36 were synergy on regulating inflammation factors expression. This study will provide a new theory basis for mechanism research of ALI with sepsis, and has a very important biological significance on treatment of ALI with sepsis. |
PDF Full Text Request