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Hepatocyte-Derived Exosomal microRNAs Orchestrate Vascular Inflammation and Endothelial Function: Insights Into Molecular Mechanisms of Trimethylamine-N-Oxide in Atherosclerosis
Jimei Chen
Department of Cardiovascular Surgery, Guangdong Cardiovascular Institute, Guangdong Provincial Key Laboratory of South China Structural Heart Disease,Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences
Corresponding Author
ORCiD: https://orcid.org/0000-0002-0207-2345
License:
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Background
Trimethylamine-N-oxide (TMAO) has been proved to be a new proatherogenic compound for promoting vascular inflammation and endothelial dysfunction. Hepatocyte-derived exosomes played an important role in the regulation of vascular inflammation and endothelial function. Since TMAO is produced in the liver, hepatocytes may be the first potential target of TMAO. However, it is not clear whether TMAO can directly stimulate normal hepatocytes to produce exosomes so as to mediate the motivating effects of TMAO on inflammation and endothelial dysfunction.
Methods
The hepatocytes were cultured and treated with TMAO at a physiological concentration for 24 hours (TMAO-Exos). The untreated group served as the control (Control-Exos). The exosomes were isolated from the culture supernatant and then added to the human aortic endothelial cells (HAECs) for 48 hours. The mRNA expressions of inflammatory cytokines and caspase-3 were determined by qPCR and cell apoptosis was evaluated by using the Hoechst 33342 staining solution. The miRNA profile in the exosomes were detected using an RNA-sequencing strategy. The miRNA-mRNA network was predicted, and the biological functions of the target genes were annotated by using bioinformatics methods.
Results
TMAO-Exos were able to promote the expressions of inflammatory cytokines and HAECs apoptosis. Moreover, miRNAs carried by the TMAO-Exos were quite different from that in the Control-Exos, including miR-92a-3p, miR-103-3p and miR-122-5p, etc. Further analysis showed that these differentially expressed miRNAs were predicted to target genes such as Mapk8, Casp9, Mapk10, Bcl2l11, Ikbkg and Akt1, which were supposed to be involved in the signal pathways related to vascular inflammation and endothelial function.
Conclusions
These novel results provided evidence that TMAO could indirectly talk to vascular endothelial via promoting hepatocytes to secreting exosomes that carried important genetic information, which may give a new insight into the interactions between liver and vasculature in the atherogenesis caused by TMAO. New intervention targeting this cellular crosstalk may be feasible and effective in the prevention and treatment of TMAO-induced atherogenesis.
Keywords
Hepatocyte-derived, Vascular Inflammation, Endothelial Function, Atherosclerosis
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This preprint is under consideration at Journal of Translational Medicine. A preprint is a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research
Hepatocyte-Derived Exosomal microRNAs Orchestrate Vascular Inflammation and Endothelial Function: Insights Into Molecular Mechanisms of Trimethylamine-N-Oxide in Atherosclerosis
Jimei Chen
Department of Cardiovascular Surgery, Guangdong Cardiovascular Institute, Guangdong Provincial Key Laboratory of South China Structural Heart Disease,Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences
Corresponding Author
ORCiD: https://orcid.org/0000-0002-0207-2345
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Under Review
Version 1
Posted 22 Feb, 2021
No community comments so far
Editor assigned
On 09 Feb, 2021
Submission checks complete
On 09 Feb, 2021
Editor invited
On 09 Feb, 2021
First submitted
On 08 Feb, 2021
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Translational Medicine
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background
Trimethylamine-N-oxide (TMAO) has been proved to be a new proatherogenic compound for promoting vascular inflammation and endothelial dysfunction. Hepatocyte-derived exosomes played an important role in the regulation of vascular inflammation and endothelial function. Since TMAO is produced in the liver, hepatocytes may be the first potential target of TMAO. However, it is not clear whether TMAO can directly stimulate normal hepatocytes to produce exosomes so as to mediate the motivating effects of TMAO on inflammation and endothelial dysfunction.
Methods
The hepatocytes were cultured and treated with TMAO at a physiological concentration for 24 hours (TMAO-Exos). The untreated group served as the control (Control-Exos). The exosomes were isolated from the culture supernatant and then added to the human aortic endothelial cells (HAECs) for 48 hours. The mRNA expressions of inflammatory cytokines and caspase-3 were determined by qPCR and cell apoptosis was evaluated by using the Hoechst 33342 staining solution. The miRNA profile in the exosomes were detected using an RNA-sequencing strategy. The miRNA-mRNA network was predicted, and the biological functions of the target genes were annotated by using bioinformatics methods.
Results
TMAO-Exos were able to promote the expressions of inflammatory cytokines and HAECs apoptosis. Moreover, miRNAs carried by the TMAO-Exos were quite different from that in the Control-Exos, including miR-92a-3p, miR-103-3p and miR-122-5p, etc. Further analysis showed that these differentially expressed miRNAs were predicted to target genes such as Mapk8, Casp9, Mapk10, Bcl2l11, Ikbkg and Akt1, which were supposed to be involved in the signal pathways related to vascular inflammation and endothelial function.
Conclusions
These novel results provided evidence that TMAO could indirectly talk to vascular endothelial via promoting hepatocytes to secreting exosomes that carried important genetic information, which may give a new insight into the interactions between liver and vasculature in the atherogenesis caused by TMAO. New intervention targeting this cellular crosstalk may be feasible and effective in the prevention and treatment of TMAO-induced atherogenesis.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8