miR-27a is up regulated and promotes inflammatory response in sepsis

doi:10.1016/j.cellimm.2014.06.006

Cellular Immunology

Volume 290, Issue 2, August 2014, Pages 190-195

https://doi.org/10.1016/j.cellimm.2014.06.006 Get rights and content

Highlights

•
miR-27a is up regulated in sepsis.
•
Inhibition of miR-27a reduces the expression levels of TNF-α and IL-6.
•
Knocking down of miR-27a reduces the phosphorylation level of NF-κB p65 and inhibits its DNA binding activity.
•
Neutralisation of miR-27a relieves pulmonary inflammation and increases survival rate of septic mice.

Abstract

MicroRNAs (miRNAs) are short, non-coding RNAs that regulate the expression of multiple target genes. Dysregulation of miRNAs is common in sepsis. Through microRNA microarray and qRT-PCR we found that the levels of miR-27a, miR-153 and miR-143 are up regulated, while let-7a, miR-218 and miR-129-5p are down regulated in lungs of septic mice. Knocking down of miR-27a down regulates expression levels of TNF-α and IL-6 significantly via reducing the phosphorylation level of NF-κB p65 and inhibiting its DNA binding activity. Furthermore, neutralisation of miR-27a up regulates PPARγ level, down regulates TNF-α expression, relieves pulmonary inflammation and promotes survival of septic mice, which demonstrates that miR-27a plays an important role in regulating inflammatory response in sepsis and provides a potential target for clinical sepsis research and treatment.

Introduction

Despite of the rapid progression of medical care, sepsis remains to be the leading cause of death in intensive care units [1]. Sepsis is caused by the immune system’s response to a serious infection, most commonly bacteria, but also fungi, viruses, and parasites in the blood, urinary tract, lungs, skin, or other tissues [2]. Sepsis can be thought of as falling within a continuum from infection to multiple organ dysfunction syndrome, including acute lung injury (ALI) [3].

Recently, several studies have reported that NF-κB plays an important role in sepsis and LPS may active NF-κB through Toll-like pathway [4]. The NF-κB family has five members: p105/p50 (NF-κB1), p100/p52 (NF-κB2), p65 (RelA), RelB and c-Rel. After stimulation, IKKα and IKKβ are phosphorylated, promoting the phosphorylation of IκBα, which promotes its degradation via the ubiquitin–proteasome pathway [5]. The degradation of IκBα leads to NF-κB p50/NF-κB p65 dimer phosphorylation, nuclear translocation and transcription of downstream target genes, including genes encoding inflammatory cytokines [6].

Peroxisome proliferator-activated receptors are ligand-activated transcription factors belonging to the nuclear hormone receptor family [7], among which, PPARα (also known as NR1C3), γ (NR1C1) and δ (NR1C2), are widely expressed and have a wide range of effects on metabolism, cell proliferation and immune responses [8]. PPARγ was recently identified as a potent modulator of inflammation, it can inhibit NF-κB, leading to suppression of downstream targets, including the genes encoding proinflammatory cytokines [9], [10].

MicroRNAs (miRNAs) are critical regulators of gene expression. Mature miRNAs bind to target mRNAs at complementary sites in 3′-untranslated regions (3′-UTRs) or coding sequences and thereby trigger down-regulation of target genes expression [11]. This study was designed to determine the expression regulation and the underlying function of microRNAs in sepsis and the pathogenesis of ALI. Through microRNA microarray and qRT-PCR we found that the levels of miR-27a, miR-153 and miR-143 are up regulated, while let-7a, miR-218 and miR-129-5p are down regulated in lungs of septic mice. Knocking down of miR-27a down regulates expression levels of TNF-α and IL-6 significantly via reducing the phosphorylation level of NF-κB p65 and inhibiting its DNA binding activity. Furthermore, neutralisation of miR-27a up regulates PPARγ level, relieves pulmonary inflammation and promotes survival of septic mice, which demonstrates that miR-27a plays an important role in regulating inflammatory response in sepsis and provides a potential target for clinical sepsis research and treatment.

Section snippets

Animals and CLP model of sepsis

Healthy 7-week-old C57BL/6 male mice, weight 22–25 g, were purchased from the Animals Experimentation Center of Fudan University. All animals were kept under controlled temperature (20 ± 2 °C), humidity (60 ± 5%) and 12 h light/12 h dark cycle for 1 week before the experiment. The CLP procedure was performed according to the guidelines of Remick et al. [12]. Briefly, the CLP model of sepsis is induced by laparotomy and exposure of the junction between the large and small intestines. The small intestine

Sepsis causes pulmonary inflammation in vivo

To assess pulmonary pathology of the CLP induced mice model of sepsis, histological evaluation was used. Lung sections 3 day after the CLP displayed notable pathological feature: inflammatory cells infiltration, interstitial edema, inter-alveolar septal thickening (Fig. 1B), while the lung sections from control group showed no histological changes (Fig. 1A). Moreover, the lung wet to dry weight ratio (W/D) was 3.75 ± 0.43 and 7.08 ± 1.03 in normal and septic mice, respectively (Fig. 1C), and the MPO

Discussion

Sepsis is a potentially fatal whole-body inflammation (a systemic inflammatory response syndrome, SIRS) caused by severe infection [18]. Severe sepsis is a complicated organ dysfunction [19] and acute lung injury (ALI) is a common complication of sepsis [20]. This study was aimed to find out the function of microRNAs which were up regulated in sepsis and the effect of knocking down the microRNA on sepsis pathology and ALI.

We made septic mice with CLP model. The H&E staining, W/D ratio and MPO

Acknowledgments

This work was supported by National Natural Science Foundation of China (No. 81272144), Science and Technology Commission of Shanghai Municipality (No. 12ZR1419800).

References (22)

Y. Wang et al.
Transcriptional regulation of co-expressed microRNA target genes
Genomics
(2011)
S.Y. Kim
miR-27a is a negative regulator of adipocyte differentiation via suppressing PPARgamma expression
Biochem. Biophys. Res. Commun.
(2010)
S.M. Gutierrez
Candidemia in ICU patients with sepsis
Crit. Care Med.
(2013)
A. Panagiotou et al.
Extracorporeal therapies in sepsis
J. Intensive Care Med.
(2013)
Z. Huang
Effect on extrapulmonary sepsis-induced acute lung injury by hemoperfusion with neutral microporous resin column
Ther. Apher. Dial.
(2013)
H. Li
NF-kappaB inhibition after cecal ligation and puncture reduces sepsis-associated lung injury without altering bacterial host defense
Mediators Inflamm.
(2013)
T. Zhu
Andrographolide protects against LPS-induced acute lung injury by inactivation of NF-kappaB
PLoS One
(2013)
C. Meyer
Transcript and protein profiling identifies signaling, growth arrest, apoptosis, and NF-kappaB survival signatures following GNRH receptor activation
Endocr. Relat. Cancer
(2013)
X.Y. Zhang et al.
PPAR family and its relationship to metabolic syndrome
Sheng Li Ke Xue Jin Zhan
(2005)
J. Kamon et al.
PPAR family (PPAR alpha, PPAR delta, PPAR gamma)
Nihon Rinsho
(2002)

W. Wahli

A gut feeling of the PXR, PPAR and NF-kappaB connection

J. Intern. Med.

(2008)

Cited by (55)

lncRNA NEAT1 aggravates sepsis-induced lung injury by regulating the miR-27a/PTEN axis
2021, Laboratory Investigation
Sepsis is an acute inflammatory reaction and a cause of acute respiratory distress syndrome (ARDS). In the present study, we explored the roles and underlying mechanism of the lncRNA Nuclear enriched abundant transcript 1 (NEAT1) in ARDS. The expression levels of genes, proteins and pro-inflammatory cytokines in patients with ARDS, LPS-stimulated cells and septic mouse models were quantified using qPCR, western blotting and ELISA assays, respectively. The molecular targeting relationship was validated by conducting a dual-luciferase reporter assay. Cell proliferation was assessed using the Cell Counting Kit-8 (CCK-8) assay. The cell cycle phase was determined by flow cytometry assay. The expression levels of NEAT1 and pro-inflammatory cytokines were higher in patients with ARDS and septic models than in controls. Knockdown of NEAT1 significantly increased cell proliferation and cycle progression and prolonged mouse survival in vitro and in vivo. Mechanistically, miR-27a was identified as a downstream target of NEAT1 and directly inhibited PTEN expression. Further rescue experiments revealed that inhibition of miR-27a impeded the promoting effects of NEAT1 silence on cell proliferation and cycle progression, whereas inhibition of PTEN markedly weakened the inhibitory effects of NEAT1 overexpression on cell proliferation and cycle progression. Altogether, our study revealed that NEAT1 plays a promoting role in the progression of ARDS via the NEAT1/miR-27a/PTEN regulatory network, providing new insight into the pathologic mechanism behind ARDS.
Sepsis-induced acute lung injury has been clinically defined as acute respiratory distress syndrome (ARDS). In the present study, the authors reveal that NEAT1 plays a role in the inflammation and cell cycle progression of ARDS via the NEAT1/miR-27a/PTEN regulatory network, providing new insight into the pathologic mechanism behind ARDS.
miR-129-5p alleviates LPS-induced acute kidney injury via targeting HMGB1/TLRs/NF-kappaB pathway
2020, International Immunopharmacology
The present study aimed to investigate whether miR-129-5p can regulate high-mobility group box protein 1 (HMGB1)-modulated TLRs/NF-kappaB inflammatory pathway that contributed to lipopolysaccharide (LPS)-induced podocyte apoptosis and acute kidney injury (AKI).
In vitro and in vivo models of sepsis were simulated using LPS-administrated podocytes and mice, respectively. The effects of LPS, mR-129-5p mimics and short hairpin RNA of HMGB1 (sh-HMGB1) on podocyte apoptosis were monitored using TUNEL staining. Protein expression was measured using western blotting. Survival outcomes were analyzed in septic mice with agomir-mR-129-5p administration.
We observed that stimulation of podocytes with LPS significantly inhibits the expression of miR-129-5p, and overexpression of miR-129-5p protects against LPS-induced podocyte damage, over-activation of inflammatory response and apoptosis. In a mouse model, agomir-miR-129-5p administration significantly improves the survival outcomes in septic mice and LPS-induced AKI. Mechanically, LPS-induced the elevation of HMGB1, TLR2, TLR4 and nuclear NF-κB protein expression in vitro and in vivo are restrained by the overexpression of miR-129-5p.
Overexpression of miR-129-5p protects against LPS-induced podocyte apoptosis, inflammation and AKI in vivo and in vitro models of sepsis. The underlying molecular mechanism is mediated via attenuating HMGB1/TLRs/NF-κB signaling axis modulated inflammatory response.
TUG1 Represses Apoptosis, Autophagy, and Inflammatory Response by Regulating miR-27a-3p/SLIT2 in LPS-Treated Vascular Endothelial Cells
2020, Journal of Surgical Research
Citation Excerpt :
These reports indicated the proinflammatory role of miR-27a-3p in multiple cell lines. Here, we found that miR-27a-3p level was increased in LPS-treated HUVECs, and miR-27a-3p knockdown inhibited LPS-induced injury, which was also consistent with that in the former work.15 In addition, miR-27a-3p overexpression reversed TUG1-mediated inhibitive role in LPS-induced injury, implying that TUG1 could suppress sepsis development by sponging miR-27a-3p.
The dysfunction of vascular endothelial cells is associated with sepsis development. Long noncoding RNAs take part in the regulation of vascular endothelial cell function. This study aimed to explore the role and mechanism of long noncoding RNA taurine-upregulated gene 1 (TUG1) in lipopolysaccharide (LPS)-induced endothelial cell injury.
LPS-treated human umbilical vein endothelial cells (HUVECs) were used as a model of sepsis in vitro. Quantitative real-time polymerase chain reaction was performed to detect the expression of TUG1, microRNA-27a-3p (miR-27a-3p) and slit guidance ligand 2 (SLIT2) messenger RNA. Western blot was conducted to measure the protein levels of SLIT2 as well as those involved in apoptosis, autophagy, and inflammatory response. Flow cytometry was used to detect cell apoptotic rate. The targets of TUG1 and miR-27a-3p were predicted via starBase (http://starbase.sysu.edu.cn/index.php). Dual-luciferase reporter, RNA immunoprecipitation, and pull-down assays were carried out to validate the target correlation between miR-27a-3p and TUG1/SLIT2.
TUG1 expression was decreased after the treatment of LPS in HUVECs. Overexpression of TUG1 decreased LPS-induced apoptosis, autophagy, and inflammatory response. TUG1 was a sponge of miR-27a-3p. Upregulation of miR-27a-3p reversed the suppressive effect of TUG1 overexpression on LPS-induced apoptosis, autophagy, and inflammatory response. SLIT2 was a target of miR-27a-3p. Knockdown of miR-27a-3p could inhibit LPS-induced injury by increasing SLIT2 in HUVECs. TUG1 could enhance SLIT2 expression by competitively sponging miR-27a-3p.
TUG1 could repress cell apoptosis, autophagy, and inflammatory response in LPS-treated HUVECs by sponging miR-27a-3p to target SLIT2, providing a potential target for the treatment of sepsis.
Overexpression of miR-129-5p Mitigates Sepsis-Induced Acute Lung Injury by Targeting High Mobility Group Box 1
2020, Journal of Surgical Research
Citation Excerpt :
The former efforts demonstrate that miR-129-5p could exhibit the protective effect in various disorders by inhibiting the progression of cancers and diseases.9-11 More importantly, miR-129-5p expression is dysregulated in lung tissues of mice with sepsis.12 This report implies that miR-129-5p might be involved in sepsis-induced acute lung injury.
MicroRNAs are dysregulated in sepsis. Acute lung injury is a progressive syndrome during sepsis. However, the role of miR-129-5p in the development of acute lung injury induced by sepsis remains unclear.
The acute lung injury of sepsis model was established by cecal ligation puncture (CLP)-treated mice and lipopolysaccharide (LPS)-treated murine alveolar epithelial cell line (MLE)-12 cells. The lung injury in vivo was investigated by hematoxylin and eosin staining, terminal dexynucleotidyl transferase (TdT)-mediated dUTP nick end labeling staining, enzyme-linked immunosorbent assay, lung wet-to-dry weight ratio, and myeloperoxidase activity. The lung injury in vitro was evaluated by 3-(4,5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide, flow cytometry, and enzyme-linked immunosorbent assay. The expression levels of miR-129-5p and high mobility group box 1 (HMGB1) were measured by quantitative real-time polymerase chain reaction and Western blot. The association between miR-129-5p and HMGB1 was validated by luciferase assay and RNA immunoprecipitation.
The expression of miR-129-5p was decreased in CLP model and LPS-treated MLE-12 cells. Overexpression of miR-129-5p attenuated inflammatory response, apoptosis, lung wet/dry weight ratio, and myeloperoxidase activity induced by CLP surgery in vivo. Moreover, addition of miR-129-5p increased cell viability and suppressed cell apoptosis and inflammatory response in vitro. HMGB1 as a target of miR-129-5p alleviated miR-129-5p–mediated injury suppression in LPS-treated MLE-12 cells.
miR-129-5p protects against sepsis-induced acute lung injury by decreasing HMGB1 expression, providing new target for sepsis treatment.
Molecular mechanisms of organ damage in sepsis: an overview
2020, Brazilian Journal of Infectious Diseases
Citation Excerpt :
These studies have shown that miRNA expression occurs in the course of various pathological infection processes [61]. It has also been shown that the deregulation of miRNA expression correlates with symptoms of severe inflammation as well as sepsis [62–66]. In the course of sepsis, also other miRNA molecules, including miRNA-25, miRNA-133a, miRNA-146, miRNA-150 and miRNA-223 are deregulated and thus they may be used as markers in different stages of sepsis [59,72].
Sepsis is one of the most common reasons for hospitalization. This condition is characterized by systemic inflammatory response to infection. International definition of sepsis mainly points out a multi-organ dysfunction caused by a deregulated host response to infection. An uncontrolled inflammatory response, often referred to as “cytokine storm”, leads to an increase in oxidative stress as a result of the inhibition of cellular antioxidant systems. Oxidative stress, as well as pro-inflammatory cytokines, initiate vascular endothelial dysfunction and, in consequence, impair microcirculation. Microcirculation damage leads to adaptive modifications of cell metabolism. Moreover, mitochondrial dysfunction takes place which results in increased apoptosis and organ damage. Non-coding RNA fragments, especially miRNA molecules, may play an important role in the pathomechanism of sepsis. Numerous studies have indicated altered expression of various miRNAs in sepsis. miRNAs can be used as markers in the diagnosis and prognosis of disease development. In turn, intracellular miRNAs regulate the TLR4/NFκB pathway responsible for the expression of pro-inflammatory cytokine genes involved in the inflammatory response in sepsis. The understanding of detailed molecular mechanisms leading to organ damage can contribute to the development of specific therapy methods thereby improving the prognosis of patients with sepsis.
Non-coding RNAs and Exosomes: Their Role in the Pathogenesis of Sepsis
2020, Molecular Therapy Nucleic Acids
Sepsis is characterized as an uncontrolled host response to infection, and it represents a serious health challenge, causing excess mortality and morbidity worldwide. The discovery of sepsis-related epigenetic and molecular mechanisms could result in improved diagnostic and therapeutic approaches, leading to a reduced overall risk for affected patients. Accumulating data show that microRNAs, non-coding RNAs, and exosomes could all be considered as novel diagnostic markers for sepsis patients. These biomarkers have been demonstrated to be involved in regulation of sepsis pathophysiology. However, epigenetic modifications have not yet been widely reported in actual clinical settings, and further investigation is required to determine their importance in intensive care patients. Further studies should be carried out to explore tissue-specific or organ-specific epigenetic RNA-based biomarkers and their therapeutic potential in sepsis patients.

View all citing articles on Scopus

¹: Zhongchuan Wang and Zhengshang Ruan contributed equally to this work, are co-first authors.

View full text

Rapid CommunicationmiR-27a is up regulated and promotes inflammatory response in sepsis

Highlights

Abstract

Introduction

Section snippets

Animals and CLP model of sepsis

Sepsis causes pulmonary inflammation in vivo

Discussion

Acknowledgments

Genomics

Biochem. Biophys. Res. Commun.

Candidemia in ICU patients with sepsis

Crit. Care Med.

Extracorporeal therapies in sepsis

J. Intensive Care Med.

Effect on extrapulmonary sepsis-induced acute lung injury by hemoperfusion with neutral microporous resin column

Ther. Apher. Dial.

NF-kappaB inhibition after cecal ligation and puncture reduces sepsis-associated lung injury without altering bacterial host defense

Mediators Inflamm.

Andrographolide protects against LPS-induced acute lung injury by inactivation of NF-kappaB

PLoS One

Transcript and protein profiling identifies signaling, growth arrest, apoptosis, and NF-kappaB survival signatures following GNRH receptor activation

Endocr. Relat. Cancer

PPAR family and its relationship to metabolic syndrome

Sheng Li Ke Xue Jin Zhan

PPAR family (PPAR alpha, PPAR delta, PPAR gamma)

Nihon Rinsho

A gut feeling of the PXR, PPAR and NF-kappaB connection

J. Intern. Med.

Rapid Communication
miR-27a is up regulated and promotes inflammatory response in sepsis