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. 2018 Dec;16(6):4836-4842.
doi: 10.3892/etm.2018.6779. Epub 2018 Sep 19.

Regulatory role of miRNA-26a in neonatal sepsis

Qi Cheng  1   2 , Lili Tang  3 , Yibiao Wang  4
Affiliations

Regulatory role of miRNA-26a in neonatal sepsis

Qi Cheng et al. Exp Ther Med. 2018 Dec.

Abstract

The present study aimed to investigate the expression of microRNA (miRNA) 26a in blood mononuclear cells and serum in neonatal sepsis, as well as its role in the disease pathogenesis. In total 28 cases of neonatal sepsis were included in the study. The mRNA expression levels of miRNA-26a and interleukin (IL)-6 in the blood mononuclear cells and serum samples were detected by reverse transcription-quantitative polymerase chain reaction. The protein expression of IL-6 was detected by western blot analysis and ELISA. The in vitro septic environment was simulated by lipopolysaccharide (LPS) in THP-1 cells, and the expression of miRNA-26a and IL-6 were determined. Interaction between miRNA-26a and IL-6 was confirmed by a dual-luciferase reporter assay. Compared with the control group, the mRNA and protein expression levels of IL-6 in the blood mononuclear cells and serum samples from the neonates with sepsis were significantly elevated, while the expression of miRNA-26a was significantly decreased. In addition, similar results were observed in the LPS-induced septic models in THP-1 cells. Furthermore, the results of the dual-luciferase reporter assay demonstrated that IL-6 was the direct target of miRNA-26a. The expression of IL-6 was significantly upregulated in the blood mononuclear cells and serum in neonatal sepsis, which may be associated with the downregulation of miRNA-26a. miRNA-26a may regulate the disease pathogenesis and immune responses.

Keywords: THP-1 cells; interleukin-6; microRNA-26a; neonatal sepsis.

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Figures

Figure 1.
Figure 1.
mRNA expression levels of IL-6 in blood mononuclear cells and serum samples in neonatal sepsis. Reverse transcription-quantitative polymerase chain reaction was performed to detect the mRNA expression levels of IL-6 in the (A) blood mononuclear cells and (B) serum samples in the cases of neonatal sepsis. **P<0.01 vs. the control group. IL, interleukin.
Figure 2.
Figure 2.
Protein expression levels of IL-6 in blood mononuclear cells and serum samples in neonatal sepsis. Western blot analysis and ELISA were performed to detect the protein expression of IL-6 in the (A) blood mononuclear cells and (B) serum samples, respectively in the neonatal sepsis. *P<0.05 and **P<0.01 vs. the control group. IL, interleukin; ELISA, enzyme-linked immunosorbent assay.
Figure 3.
Figure 3.
Bioinformatics analysis. The miRNA-26a was predicted as the up-stream regulator of IL-6 according to the bioinformatics analysis. IL, interleukin; miRNA, microRNA.
Figure 4.
Figure 4.
Expression levels of miRNA-26a in blood mononuclear cells and serum samples in neonatal sepsis. Reverse transcription-quantitative polymerase chain reaction was performed to detect the expression levels of miRNA-26a in the (A) blood mononuclear cells and (B) serum samples in the cases of neonatal sepsis. **P<0.01 vs. the control group. miRNA, microRNA.
Figure 5.
Figure 5.
Expression levels of IL-6 and miRNA-26a in LPS-induced THP-1 cells. THP-1 cells were induced with LPS to simulate the in vitro septic environment. (A) The protein and (B) mRNA expression levels of IL-6 in the LPS-induced THP-1 cells were detected by RT-qPCR and western blot analysis, respectively. (C) The expression levels of miRNA-26a in the LPS-induced THP-1 cells were detected by RT-qPCR. **P<0.01 vs. the control group. RT-qPCR, reverse transcription-quantitative polymerase chain reaction; IL, interleukin; LPS, lipopolysaccharide; miRNA, microRNA.
Figure 6.
Figure 6.
Dual-luciferase reporter assay. Direct interaction between miRNA-26a and IL-6 was confirmed by a dual-luciferase reporter assay. **P<0.01 vs. the NC group. NC, negative control; miRNA, microRNA; IL, interleukin.
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