This site needs JavaScript to work properly. Please enable it to take advantage of the complete set of features!
Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

NIH NLM Logo
Log in
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Comparative Study
. 2017 Jun 29;12(6):e0179850.
doi: 10.1371/journal.pone.0179850. eCollection 2017.

miR-146a, miR-146b, and miR-155 increase expression of IL-6 and IL-8 and support HSP10 in an In vitro sepsis model

Affiliations
Comparative Study

miR-146a, miR-146b, and miR-155 increase expression of IL-6 and IL-8 and support HSP10 in an In vitro sepsis model

Dagmar Pfeiffer et al. PLoS One. .

Abstract

microRNAs (miRNAs) play an essential role in inflammation processes including sepsis. This study aimed to identify miRNAs as candidates for therapies that are involved in the innate immune response and to assess their potential functions in the activation of the endothelium. We stimulated THP-1 monocytes with 10 ng/ml LPS for 4 h and used the supernatant for the stimulation of human umbilical vein endothelial cells (HUVEC) or human pulmonary microvascular endothelial cells (HPMEC) for 16 h. miRNA array analysis (of 1,891 miRNAs) identified a 1.5-fold upregulation of miR-146a, miR-146b, and miR-155 in stimulated endothelial cells. HUVEC were transfected with miRNA inhibitors for miR-146a, miR-146b, and miR-155 to investigate the function of these miRNAs in endothelial inflammatory pathways. Inhibition of miR-146a resulted in a diminished release of interleukin (IL)-6 and IL-8 by respective 68% and 55% (P<0.001). Inhibition of miR-146b reduced the expression of IL-6 by 49% (P<0.001). Inhibition of miR-155 reduced the expression of IL-6 and IL-8 by respective 31% (P<0.001) and 14%. The inhibition of miR-146a, miR-146b, and miR-155 reduced the release of HSP10 by 50%, 35%, and 69% (P<0.05), respectively, but did not influence the expression of HSP27 or TXA2. In conclusion, miR-146a, miR-146b, and miR-155 are exerting anti-inflammatory properties by down-regulating IL-6 and IL-8, and influencing the expression of HSP10 in the activated endothelium. We provide evidence for the central role of selected miRNAs in sepsis and their use in the development of small interfering RNA therapeutics to target immune cells and sepsis pathways.

PubMed Disclaimer

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Fig 1
Fig 1. Cytokine profile of stimulated THP-1.
THP-1 monocytes were stimulated with 10 ng/ml LPS, and expression of TNF-α, IL-1β, IL-6, IL-8, and IL-10 was measured after 4 h. The supernatant was collected and used as CdM for stimulation of HUVEC. The data represent the means ± SD (n = 3). **P < 0.001.
Fig 2
Fig 2. Cytokine profile of CdM-stimulated HUVEC.
HUVEC were stimulated with CdM and expression of TNF-α, IL-1β, IL-6, IL-8, and IL-10 was measured after 16 h. The data represent the means ± SD (n = 3). **P < 0.001.
Fig 3
Fig 3. Different expression of IL-6 and IL-8 of HUVEC stimulated with CdM compared to TNF-α.
HUVEC were stimulated with CdM or 800 pg TNF-α /ml, and expression of IL-6 (A) and IL-8 (B) was measured after 4 h, 6 h, 16 h, and 24 h. The data represent the means ± SD (n = 3).
Fig 4
Fig 4. miRNA expression changes in HUVEC and HPMEC stimulated with CdM.
We stimulated HUVEC (A) and HPMEC (B) with CdM for 16 h and investigated 1,891 miRNAs by array analysis. The bars indicate fold changes in miRNA expression (delta LMR) compared to the unstimulated control. The data represent the means ± SD (n = 3).
Fig 5
Fig 5. Changes in the expression of IL-6 and IL-8 in CdM-stimulated and miR-146a-, miR-146b- or miR-155-inhibited HUVEC.
We stimulated HUVEC with CdM after inhibition of miR-146a, miR-146b or miR-155 and measured the expression of IL-6 and IL-8 after 16 h. We added inhibitors for miR-146a, miR-146b or miR-155 at concentrations of 25 mM or 50 mM. A mock control miRNA served as the control. The data represent the means ± SD (n = 3). **P < 0.001.
Fig 6
Fig 6. Inhibition of miR-146a, miR-146b, and miR-155 reduced the expression of HSP10 in CdM-stimulated HUVEC.
We stimulated HUVEC with CdM after inhibition of miR-146a, miR-146b, or miR-155 and measured the expression of HSP10 after 16 h. CdM-stimulated HUVEC acted as a control, and their expression levels were set to 100%. The data represent the means ± SD (n = 3). *P < 0.05.

References

    1. Singer M, Deutschman CS, Seymour CW, Shankar-Hari M, Annane D, et al. (2016) The Third International Consensus Definitions for Sepsis and Septic Shock (Sepsis-3). JAMA 315: 801–810. doi: 10.1001/jama.2016.0287 - DOI - PMC - PubMed
    1. Ince C, Mik EG (2016) Microcirculatory and mitochondrial hypoxia in sepsis, shock, and resuscitation. J Appl Physiol (1985) 120: 226–235. - PubMed
    1. Henderson B (2010) Integrating the cell stress response: a new view of molecular chaperones as immunological and physiological homeostatic regulators. Cell Biochem Funct 28: 1–14. doi: 10.1002/cbf.1609 - DOI - PubMed
    1. Giuliano JS Jr., Lahni PM, Wong HR, Wheeler DS (2011) Pediatric Sepsis—Part V: Extracellular Heat Shock Proteins: Alarmins for the Host Immune System. Open Inflamm J 4: 49–60. doi: 10.2174/1875041901104010049 - DOI - PMC - PubMed
    1. Johnson BJ, Le TT, Dobbin CA, Banovic T, Howard CB, et al. (2005) Heat shock protein 10 inhibits lipopolysaccharide-induced inflammatory mediator production. J Biol Chem 280: 4037–4047. doi: 10.1074/jbc.M411569200 - DOI - PubMed

Publication types

Full text links
Public Library of Science full text link Public Library of Science Free PMC article
Cite

AltStyle によって変換されたページ (->オリジナル) /