Analysis of miRNA expression profile in lung tissues of an intermittent hypoxia rat model
Introduction
Obstructive sleep apnea (OSA), the clinical features of which include snoring, apnea, and daytime sleepiness, is characterized by apnea and hypopnea disorder caused by the collapse of the upper airway during sleep (Lévy et al., 2015). In particular, OSA can be complicated by hypertension, diabetes, coronary heart disease, arrhythmia, cerebral apoplexy, cognitive dysfunction, and other chronic diseases (Jordan et al., 2014). Recent studies have revealed that OSA is related to the incidence and mortality of tumors (Nieto et al., 2012). The incidence rate of OSA has been reported to be 14 % and 5 % in American adult males and females, respectively (Peppard et al., 2013). Latest research has shown that currently 936 million adults aged between 30–69 years are suffering from OSA, with China having the highest prevalence of OSA worldwide (Benjafield et al., 2019).
Chronic intermittent hypoxia (CIH) is known to be the characteristic pathophysiological change associated with OSA. The process of hypoxia/reoxygenation, which is similar to ischemia/reperfusion injury, has been shown to produce excessive reactive oxygen species (ROS), resulting in the imbalance of oxidants and antioxidants in the body (Lavie, 2015; Lui et al., 2013). Moreover, ROS are known to activate the "inflammatory cascade", leading to the increased expression of pro-inflammatory cytokines and adhesion molecules (Pilkauskaite et al., 2013), which ultimately lead to systemic multi-organ function impairment.
MicroRNAs (miRNAs) are small non-coding RNAs (20−22‒nucleotide long) that bind to the 3′ untranslated regions (3′-UTR) of mRNAs to negatively regulate the expression of target genes (Bartoszewski and Sikorski, 2018). The expression of miRNAs has been closely related to lung growth and development, maintenance of pulmonary homeostasis, and pulmonary inflammation. Hypoxic conditions have been reported to induce expression changes in some miRNAs, which are thus called " hypoxic miRNAs " (Bertero et al., 2017). It has been reported that the altered expression of miRNAs plays a role in mediating the pathophysiological changes in diseases, suggesting the use of miRNAs as valuable tools for the diagnosis, treatment, and prognosis of these diseases (Luo et al., 2017; Yu et al., 2019).
In this study, we established rat models with different levels of intermittent hypoxia to simulate patients with varying severities of OSA, and used miRNA chip technology to detect the expression profile of miRNAs in rat lung tissues and explore the function of miRNAs in the context of OSA development.
Section snippets
Animal model
In our study, animal experiments complied with the ARRIVE guidelines and carried out in accordance with the U.K. Animals (Scientific Procedures) Act, 1986 and associated guidelines, EU Directive 2010/63/EU for animal experiments and Laboratory Animal Care and Use Standard of Kunming Medical University(Kunming, China). Animal experimental were approved by the Ethics Committee of Kunming Medical University. Briefly, 50 adult male Wistar rats (HFK Bioscience Co., LTD, Beijing, China), which with
Expressed miRNAs among different groups
We found that the expression of 47 miRNAs in the lung tissues of the IH1 group was significantly different (P < 0.05) compared with that of the control group. More specifically, 24 miRNAs were demonstrated to be upregulated, whereas 23 miRNAs were downregulated (Fig. 1A). A similar trend was observed in the lung tissues of the IH2 group (47 miRNAs, P < 0.05), in which 19 miRNAs were upregulated, whereas 28 miRNAs were downregulated (Fig. 1B). We also observed that 21 miRNAs were upregulated,
Discussion
Obstructive sleep apnea is a systemic disease of complex etiology. MiRNAs are important molecules, which regulate both the transcriptional and post-transcriptional processes, and play an important role in regulating cell growth, differentiation, stress, and apoptosis (Bartel, 2004). In this study, an miRNA microarray was used to screen the differentially expressed miRNAs in the lung tissues of different intermittent hypoxia groups and a negative control group.
We identified differences in the
Conclusions
In summary, we screened miRNAs in an intermittent hypoxia rat model and explored the biological roles of downstream target genes and related regulatory pathways. The aggravation of hypoxia corresponded with an altered expression profile of miRNAs. The expression profile of miRNAs in the lung tissues of rats exposed to different hypoxia levels identified certain common and differentially expressed miRNAs was detected. Some miRNAs had excellent diagnostic value for detecting OSA. The LKB1/AMPK
Author contributions
Zhijuan Liu: Conceptualization, Methodology, Formal analysis, Investigation, Writing - Original Draft, Writing - Review & Editing. Li Ai: Conceptualization, Methodology, Formal analysis, Investigation. Ran Li: Conceptualization, Methodology. Yuan Yang: Methodology, Formal analysis. Keli Chen: Methodology, Investigation. Chunxia He: Conceptualization, Methodology, Formal analysis. Yongxia Li: Conceptualization, Methodology, Writing - Review & Editing.
Funding
This work was supported by the National Natural Science Foundation of China (grant number: 81660019).
Data statement
All data, materials and software application support the published claims and comply with field standards.
Declaration of Competing Interest
The authors declare that they have no conflict of interest.
Acknowledgements
This work was supported by the National Natural Science Foundation of China (grant number: 81660019).
References (46)
MicroRNAs: genomics, biogenesis, mechanism, and function
Cell
(2004)- et al.
Estimation of the global prevalence and burden of obstructive sleep apnoea: a literature-based analysis
Lancet Respir. Med.
(2019) - et al.
Impact of MicroRNAs in the cellular response to hypoxia
Int. Rev. Cell Mol. Biol.
(2017) - et al.
Integrative genomic analysis reveals a high frequency of LKB1 genetic alteration in Chinese lung adenocarcinomas
J. Thorac. Oncol.
(2014) - et al.
Identification of the sucrose non- fermenting related kinase SNRK, as a novel LKB1 substrate
FEBS Lett.
(2005) - et al.
Adult obstructive sleep apnoea
Lancet
(2014) - et al.
miRNA networks modulate human endothelial cell adaptation to cyclic hypoxia
Cell. Signal.
(2019) Oxidative stress in obstructive sleep apnea and intermittent hypoxia--revisited--the bad ugly and good: implications to the heart and brain
Sleep Med. Rev.
(2015)- et al.
The induction of reactive oxygen species and loss of mitochondrial Omi/HtrA2 is associated with S-nitrosoglutathione-induced apoptosis in human endothelial cells
Toxicol. Appl. Pharmacol.
(2010) - et al.
MicroRNAs to guide medical decision-making in obstructive sleep apnea: a review
Sleep Med. Rev.
(2021)
Cancer risk associated with STK11/LKB1 germline mutations in Peutz-Jeghers syndrome patients: results of an Italian multicenter study
Dig. Liver Dis.
Biology and pathology of nectins and nectin-like molecules
Curr. Opin. Cell Biol.
Nectin-4 is a breast cancer stem cell marker that induces WNT/β-catenin signaling via Pi3k/Akt axis
Int. J. Biochem. Cell Biol.
A novel PI3K/AKT signaling axis mediates Nectin-4-induced gallbladder cancer cell proliferation, metastasis and tumor growth
Cancer Lett.
Editorial focus: entering into the non-coding RNA era
Cell. Mol. Biol. Lett.
The Gene Ontology (GO) project: structured vocabularies for molecular biology and their application to genome and expression analysis
Curr. Protoc. Bioinformatics
AMP-activated protein kinase and its downstream transcriptional pathways
Cell. Mol. Life Sci.
Modulation of the LKB1-AMPK signalling pathway underpins hypoxic pulmonary vasoconstriction and pulmonary hypertension
Adv. Exp. Med. Biol.
Severe obstructive sleep apnea is associated with circulating microRNAs related to heart failure, myocardial ischemia, and cancer proliferation
Sleep Breath.
MicroRNAs:small RNAs with a big role in gene regulation
Nat. Rev. Genet.
LKB1 modulates lung cancer differentiation and metastasis
Nature
Nectin-4 cis-interacts with ErbB2 and its trastuzumab-resistant splice variants, enhancing their activation and DNA synthesis
Sci. Rep.
LKB1 loss links serine metabolism to DNA methylation and tumorigenesis
Nature
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