Cargando…

MicroRNA-497 induced by Clonorchis sinensis enhances the TGF-β/Smad signaling pathway to promote hepatic fibrosis by targeting Smad7

BACKGROUND: Various stimuli, including Clonorchis sinensis infection, can cause liver fibrosis. Liver fibrosis is characterized by the activation of hepatic stellate cells (HSCs) with massive production of extracellular matrix (ECM). Our previous study showed that the TGF-β(1)-induced Smad signaling...

Descripción completa

Detalles Bibliográficos
Autores principales: Zhou, Qian-Yang, Yang, Hui-Min, Liu, Ji-Xin, Xu, Na, Li, Jing, Shen, Li-Ping, Zhang, Yu-Zhao, Koda, Stephane, Zhang, Bei-Bei, Yu, Qian, Chen, Jia-Xu, Zheng, Kui-Yang, Yan, Chao
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8442346/
https://www.ncbi.nlm.nih.gov/pubmed/34521449
http://dx.doi.org/10.1186/s13071-021-04972-3
_version_ 1783752990549606400
author Zhou, Qian-Yang
Yang, Hui-Min
Liu, Ji-Xin
Xu, Na
Li, Jing
Shen, Li-Ping
Zhang, Yu-Zhao
Koda, Stephane
Zhang, Bei-Bei
Yu, Qian
Chen, Jia-Xu
Zheng, Kui-Yang
Yan, Chao
author_facet Zhou, Qian-Yang
Yang, Hui-Min
Liu, Ji-Xin
Xu, Na
Li, Jing
Shen, Li-Ping
Zhang, Yu-Zhao
Koda, Stephane
Zhang, Bei-Bei
Yu, Qian
Chen, Jia-Xu
Zheng, Kui-Yang
Yan, Chao
author_sort Zhou, Qian-Yang
collection PubMed
description BACKGROUND: Various stimuli, including Clonorchis sinensis infection, can cause liver fibrosis. Liver fibrosis is characterized by the activation of hepatic stellate cells (HSCs) with massive production of extracellular matrix (ECM). Our previous study showed that the TGF-β(1)-induced Smad signaling pathway played a critical role in the activation of HSCs during liver fibrosis induced by worm infection; however, the mechanisms that modulate the TGF-β/Smad signaling pathway are still poorly understood. Accumulating evidence demonstrates that miRNAs act as an important regulator of activation of HSCs during liver fibrosis. METHODS: The target of miR-497 was determined by bioinformatics analysis combined with a dual-luciferase activity assay. LX-2 cells were transfected with miR-497 inhibitor and then stimulated with TGF-β(1) or excretory/secretory products of C. sinensis (CsESPs), and activation of LX-2 was assessed using qPCR or western blot. In vivo, the mice treated with CCl(4) were intravenously injected with a single dose of adeno-associated virus serotype 8 (AAV8) that overexpressed anti-miR-497 sequences or their scramble control for 6 weeks. Liver fibrosis and damage were assessed by hematoxylin and eosin (H&E) staining, Masson staining, and qPCR; the activation of the TGF-β/Smad signaling pathway was detected by qPCR or western blot. RESULTS: In the present study, the expression of miR-497 was increased in HSCs activated by TGF-β(1) or ESPs of C. sinensis. We identified that Smad7 was the target of miR-497 using combined bioinformatics analysis with luciferase activity assays. Transfection of anti-miR-497 into HSCs upregulated the expression of Smad7, leading to a decrease in the level of p-Smad2/3 and subsequent suppression of the activation of HSCs induced by TGF-β(1) or CsESPs. Furthermore, miR-497 inhibitor delivered by highly-hepatotropic (rAAV8) inhibited TGF-β/smads signaling pathway by targeting at Smad7 to ameliorate CCL4-induced liver fibrosis. CONCLUSIONS: The present study demonstrates that miR-497 promotes liver fibrogenesis by targeting Smad7 to promote TGF-β/Smad signaling pathway transduction both in vivo and in vitro, which provides a promising therapeutic strategy using anti-miR-497 against liver fibrosis. GRAPHICAL ABSTRACT: [Image: see text]
format Online
Article
Text
id pubmed-8442346
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-84423462021-09-15 MicroRNA-497 induced by Clonorchis sinensis enhances the TGF-β/Smad signaling pathway to promote hepatic fibrosis by targeting Smad7 Zhou, Qian-Yang Yang, Hui-Min Liu, Ji-Xin Xu, Na Li, Jing Shen, Li-Ping Zhang, Yu-Zhao Koda, Stephane Zhang, Bei-Bei Yu, Qian Chen, Jia-Xu Zheng, Kui-Yang Yan, Chao Parasit Vectors Research BACKGROUND: Various stimuli, including Clonorchis sinensis infection, can cause liver fibrosis. Liver fibrosis is characterized by the activation of hepatic stellate cells (HSCs) with massive production of extracellular matrix (ECM). Our previous study showed that the TGF-β(1)-induced Smad signaling pathway played a critical role in the activation of HSCs during liver fibrosis induced by worm infection; however, the mechanisms that modulate the TGF-β/Smad signaling pathway are still poorly understood. Accumulating evidence demonstrates that miRNAs act as an important regulator of activation of HSCs during liver fibrosis. METHODS: The target of miR-497 was determined by bioinformatics analysis combined with a dual-luciferase activity assay. LX-2 cells were transfected with miR-497 inhibitor and then stimulated with TGF-β(1) or excretory/secretory products of C. sinensis (CsESPs), and activation of LX-2 was assessed using qPCR or western blot. In vivo, the mice treated with CCl(4) were intravenously injected with a single dose of adeno-associated virus serotype 8 (AAV8) that overexpressed anti-miR-497 sequences or their scramble control for 6 weeks. Liver fibrosis and damage were assessed by hematoxylin and eosin (H&E) staining, Masson staining, and qPCR; the activation of the TGF-β/Smad signaling pathway was detected by qPCR or western blot. RESULTS: In the present study, the expression of miR-497 was increased in HSCs activated by TGF-β(1) or ESPs of C. sinensis. We identified that Smad7 was the target of miR-497 using combined bioinformatics analysis with luciferase activity assays. Transfection of anti-miR-497 into HSCs upregulated the expression of Smad7, leading to a decrease in the level of p-Smad2/3 and subsequent suppression of the activation of HSCs induced by TGF-β(1) or CsESPs. Furthermore, miR-497 inhibitor delivered by highly-hepatotropic (rAAV8) inhibited TGF-β/smads signaling pathway by targeting at Smad7 to ameliorate CCL4-induced liver fibrosis. CONCLUSIONS: The present study demonstrates that miR-497 promotes liver fibrogenesis by targeting Smad7 to promote TGF-β/Smad signaling pathway transduction both in vivo and in vitro, which provides a promising therapeutic strategy using anti-miR-497 against liver fibrosis. GRAPHICAL ABSTRACT: [Image: see text] BioMed Central 2021-09-14 /pmc/articles/PMC8442346/ /pubmed/34521449 http://dx.doi.org/10.1186/s13071-021-04972-3 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Zhou, Qian-Yang
Yang, Hui-Min
Liu, Ji-Xin
Xu, Na
Li, Jing
Shen, Li-Ping
Zhang, Yu-Zhao
Koda, Stephane
Zhang, Bei-Bei
Yu, Qian
Chen, Jia-Xu
Zheng, Kui-Yang
Yan, Chao
MicroRNA-497 induced by Clonorchis sinensis enhances the TGF-β/Smad signaling pathway to promote hepatic fibrosis by targeting Smad7
title MicroRNA-497 induced by Clonorchis sinensis enhances the TGF-β/Smad signaling pathway to promote hepatic fibrosis by targeting Smad7
title_full MicroRNA-497 induced by Clonorchis sinensis enhances the TGF-β/Smad signaling pathway to promote hepatic fibrosis by targeting Smad7
title_fullStr MicroRNA-497 induced by Clonorchis sinensis enhances the TGF-β/Smad signaling pathway to promote hepatic fibrosis by targeting Smad7
title_full_unstemmed MicroRNA-497 induced by Clonorchis sinensis enhances the TGF-β/Smad signaling pathway to promote hepatic fibrosis by targeting Smad7
title_short MicroRNA-497 induced by Clonorchis sinensis enhances the TGF-β/Smad signaling pathway to promote hepatic fibrosis by targeting Smad7
title_sort microrna-497 induced by clonorchis sinensis enhances the tgf-β/smad signaling pathway to promote hepatic fibrosis by targeting smad7
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8442346/
https://www.ncbi.nlm.nih.gov/pubmed/34521449
http://dx.doi.org/10.1186/s13071-021-04972-3
work_keys_str_mv AT zhouqianyang microrna497inducedbyclonorchissinensisenhancesthetgfbsmadsignalingpathwaytopromotehepaticfibrosisbytargetingsmad7
AT yanghuimin microrna497inducedbyclonorchissinensisenhancesthetgfbsmadsignalingpathwaytopromotehepaticfibrosisbytargetingsmad7
AT liujixin microrna497inducedbyclonorchissinensisenhancesthetgfbsmadsignalingpathwaytopromotehepaticfibrosisbytargetingsmad7
AT xuna microrna497inducedbyclonorchissinensisenhancesthetgfbsmadsignalingpathwaytopromotehepaticfibrosisbytargetingsmad7
AT lijing microrna497inducedbyclonorchissinensisenhancesthetgfbsmadsignalingpathwaytopromotehepaticfibrosisbytargetingsmad7
AT shenliping microrna497inducedbyclonorchissinensisenhancesthetgfbsmadsignalingpathwaytopromotehepaticfibrosisbytargetingsmad7
AT zhangyuzhao microrna497inducedbyclonorchissinensisenhancesthetgfbsmadsignalingpathwaytopromotehepaticfibrosisbytargetingsmad7
AT kodastephane microrna497inducedbyclonorchissinensisenhancesthetgfbsmadsignalingpathwaytopromotehepaticfibrosisbytargetingsmad7
AT zhangbeibei microrna497inducedbyclonorchissinensisenhancesthetgfbsmadsignalingpathwaytopromotehepaticfibrosisbytargetingsmad7
AT yuqian microrna497inducedbyclonorchissinensisenhancesthetgfbsmadsignalingpathwaytopromotehepaticfibrosisbytargetingsmad7
AT chenjiaxu microrna497inducedbyclonorchissinensisenhancesthetgfbsmadsignalingpathwaytopromotehepaticfibrosisbytargetingsmad7
AT zhengkuiyang microrna497inducedbyclonorchissinensisenhancesthetgfbsmadsignalingpathwaytopromotehepaticfibrosisbytargetingsmad7
AT yanchao microrna497inducedbyclonorchissinensisenhancesthetgfbsmadsignalingpathwaytopromotehepaticfibrosisbytargetingsmad7