Cargando…
JAK/STAT signaling pathway-mediated microRNA-181b promoted blood-brain barrier impairment by targeting sphingosine-1-phosphate receptor 1 in septic rats
BACKGROUND: Blood-brain barrier (BBB) impairment plays a significant role in the pathogenesis of sepsis-associated encephalopathy (SAE). However, the molecular mechanisms are poorly understood. In the present study, we aimed to investigate the regulatory relationship between the Janus kinase/signal...
Autores principales: | , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
AME Publishing Company
2020
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7723536/ https://www.ncbi.nlm.nih.gov/pubmed/33313203 http://dx.doi.org/10.21037/atm-20-7024 |
_version_ | 1783620359157710848 |
---|---|
author | Chen, Sheng-Long Cai, Geng-Xin Ding, Hong-Guang Liu, Xin-Qiang Wang, Zhong-Hua Jing, Yuan-Wen Han, Yong-Li Jiang, Wen-Qiang Wen, Miao-Yun |
author_facet | Chen, Sheng-Long Cai, Geng-Xin Ding, Hong-Guang Liu, Xin-Qiang Wang, Zhong-Hua Jing, Yuan-Wen Han, Yong-Li Jiang, Wen-Qiang Wen, Miao-Yun |
author_sort | Chen, Sheng-Long |
collection | PubMed |
description | BACKGROUND: Blood-brain barrier (BBB) impairment plays a significant role in the pathogenesis of sepsis-associated encephalopathy (SAE). However, the molecular mechanisms are poorly understood. In the present study, we aimed to investigate the regulatory relationship between the Janus kinase/signal transducers and activators of transcription (JAK/STAT) signaling pathway, microRNA (miR)-181b and its target genes in sepsis in vivo and in vitro. METHODS: Four rat models (sham, sepsis, sepsis plus STAT3 inhibitor (Stattic), and sepsis plus miR-181b inhibitor [sepsis + anta-miR-181b]) were established. For the in vitro experiments, rat brain microvascular endothelial cells (rBMECs) and rat brain astrocytes (rAstrocytes) were cultured with 10% serum harvested from sham, sepsis, and sepsis + anta-miR-181b rats. Chromatin immunoprecipitation-quantitative polymerase chain reaction (ChIP-QPCR) analysis was carried out to detect the binding and enrichment of the JAK/STAT3 signal core transcription complex in the miR-181b promoter region. Dual-luciferase reporter gene assay was conducted to test miR-181b and its target genes. The cell adhesion rate of rBMECs was also measured. RESULTS: During our investigations, the expression levels of miR-181b, p-JAK2, p-STAT3, and C/EBPβ were found to be significantly increased in the septic rats compared with the sham rats. STAT3 inhibitor halted BBB damage by downregulating the expression of miR-181b. In addition, miR-181b targeted sphingosine-1-phosphate receptor 1 (S1PR1) and neurocalcin delta (NCALD). The up-regulated miR-181b significantly decreased the cell adhesion rate of rBMECs. The administration of miR-181b inhibitor reduced damage to the BBB through increasing the expression of S1PR1 and NCALD, which again proved that miR-181b negatively regulates SIPR1 and NCALD to induce BBB damage. CONCLUSIONS: Our study demonstrated that JAK2/STAT3 signaling pathway induced expression of miR-181b, which promoted BBB impairment in rats with sepsis by downregulating S1PR1 and decreasing BBB cell adhesion. These findings strongly suggest JAK2/STAT3/miR-181b axis as therapeutic target in protecting against sepsis-induced BBB damage. |
format | Online Article Text |
id | pubmed-7723536 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | AME Publishing Company |
record_format | MEDLINE/PubMed |
spelling | pubmed-77235362020-12-10 JAK/STAT signaling pathway-mediated microRNA-181b promoted blood-brain barrier impairment by targeting sphingosine-1-phosphate receptor 1 in septic rats Chen, Sheng-Long Cai, Geng-Xin Ding, Hong-Guang Liu, Xin-Qiang Wang, Zhong-Hua Jing, Yuan-Wen Han, Yong-Li Jiang, Wen-Qiang Wen, Miao-Yun Ann Transl Med Original Article BACKGROUND: Blood-brain barrier (BBB) impairment plays a significant role in the pathogenesis of sepsis-associated encephalopathy (SAE). However, the molecular mechanisms are poorly understood. In the present study, we aimed to investigate the regulatory relationship between the Janus kinase/signal transducers and activators of transcription (JAK/STAT) signaling pathway, microRNA (miR)-181b and its target genes in sepsis in vivo and in vitro. METHODS: Four rat models (sham, sepsis, sepsis plus STAT3 inhibitor (Stattic), and sepsis plus miR-181b inhibitor [sepsis + anta-miR-181b]) were established. For the in vitro experiments, rat brain microvascular endothelial cells (rBMECs) and rat brain astrocytes (rAstrocytes) were cultured with 10% serum harvested from sham, sepsis, and sepsis + anta-miR-181b rats. Chromatin immunoprecipitation-quantitative polymerase chain reaction (ChIP-QPCR) analysis was carried out to detect the binding and enrichment of the JAK/STAT3 signal core transcription complex in the miR-181b promoter region. Dual-luciferase reporter gene assay was conducted to test miR-181b and its target genes. The cell adhesion rate of rBMECs was also measured. RESULTS: During our investigations, the expression levels of miR-181b, p-JAK2, p-STAT3, and C/EBPβ were found to be significantly increased in the septic rats compared with the sham rats. STAT3 inhibitor halted BBB damage by downregulating the expression of miR-181b. In addition, miR-181b targeted sphingosine-1-phosphate receptor 1 (S1PR1) and neurocalcin delta (NCALD). The up-regulated miR-181b significantly decreased the cell adhesion rate of rBMECs. The administration of miR-181b inhibitor reduced damage to the BBB through increasing the expression of S1PR1 and NCALD, which again proved that miR-181b negatively regulates SIPR1 and NCALD to induce BBB damage. CONCLUSIONS: Our study demonstrated that JAK2/STAT3 signaling pathway induced expression of miR-181b, which promoted BBB impairment in rats with sepsis by downregulating S1PR1 and decreasing BBB cell adhesion. These findings strongly suggest JAK2/STAT3/miR-181b axis as therapeutic target in protecting against sepsis-induced BBB damage. AME Publishing Company 2020-11 /pmc/articles/PMC7723536/ /pubmed/33313203 http://dx.doi.org/10.21037/atm-20-7024 Text en 2020 Annals of Translational Medicine. All rights reserved. https://creativecommons.org/licenses/by-nc-nd/4.0/Open Access Statement: This is an Open Access article distributed in accordance with the Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International License (CC BY-NC-ND 4.0), which permits the non-commercial replication and distribution of the article with the strict proviso that no changes or edits are made and the original work is properly cited (including links to both the formal publication through the relevant DOI and the license). See: https://creativecommons.org/licenses/by-nc-nd/4.0 (https://creativecommons.org/licenses/by-nc-nd/4.0/) . |
spellingShingle | Original Article Chen, Sheng-Long Cai, Geng-Xin Ding, Hong-Guang Liu, Xin-Qiang Wang, Zhong-Hua Jing, Yuan-Wen Han, Yong-Li Jiang, Wen-Qiang Wen, Miao-Yun JAK/STAT signaling pathway-mediated microRNA-181b promoted blood-brain barrier impairment by targeting sphingosine-1-phosphate receptor 1 in septic rats |
title | JAK/STAT signaling pathway-mediated microRNA-181b promoted blood-brain barrier impairment by targeting sphingosine-1-phosphate receptor 1 in septic rats |
title_full | JAK/STAT signaling pathway-mediated microRNA-181b promoted blood-brain barrier impairment by targeting sphingosine-1-phosphate receptor 1 in septic rats |
title_fullStr | JAK/STAT signaling pathway-mediated microRNA-181b promoted blood-brain barrier impairment by targeting sphingosine-1-phosphate receptor 1 in septic rats |
title_full_unstemmed | JAK/STAT signaling pathway-mediated microRNA-181b promoted blood-brain barrier impairment by targeting sphingosine-1-phosphate receptor 1 in septic rats |
title_short | JAK/STAT signaling pathway-mediated microRNA-181b promoted blood-brain barrier impairment by targeting sphingosine-1-phosphate receptor 1 in septic rats |
title_sort | jak/stat signaling pathway-mediated microrna-181b promoted blood-brain barrier impairment by targeting sphingosine-1-phosphate receptor 1 in septic rats |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7723536/ https://www.ncbi.nlm.nih.gov/pubmed/33313203 http://dx.doi.org/10.21037/atm-20-7024 |
work_keys_str_mv | AT chenshenglong jakstatsignalingpathwaymediatedmicrorna181bpromotedbloodbrainbarrierimpairmentbytargetingsphingosine1phosphatereceptor1insepticrats AT caigengxin jakstatsignalingpathwaymediatedmicrorna181bpromotedbloodbrainbarrierimpairmentbytargetingsphingosine1phosphatereceptor1insepticrats AT dinghongguang jakstatsignalingpathwaymediatedmicrorna181bpromotedbloodbrainbarrierimpairmentbytargetingsphingosine1phosphatereceptor1insepticrats AT liuxinqiang jakstatsignalingpathwaymediatedmicrorna181bpromotedbloodbrainbarrierimpairmentbytargetingsphingosine1phosphatereceptor1insepticrats AT wangzhonghua jakstatsignalingpathwaymediatedmicrorna181bpromotedbloodbrainbarrierimpairmentbytargetingsphingosine1phosphatereceptor1insepticrats AT jingyuanwen jakstatsignalingpathwaymediatedmicrorna181bpromotedbloodbrainbarrierimpairmentbytargetingsphingosine1phosphatereceptor1insepticrats AT hanyongli jakstatsignalingpathwaymediatedmicrorna181bpromotedbloodbrainbarrierimpairmentbytargetingsphingosine1phosphatereceptor1insepticrats AT jiangwenqiang jakstatsignalingpathwaymediatedmicrorna181bpromotedbloodbrainbarrierimpairmentbytargetingsphingosine1phosphatereceptor1insepticrats AT wenmiaoyun jakstatsignalingpathwaymediatedmicrorna181bpromotedbloodbrainbarrierimpairmentbytargetingsphingosine1phosphatereceptor1insepticrats |