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LncRNA np_5318 promotes renal ischemia-reperfusion injury through the TGF-β/Smad signaling pathway
Long noncoding (Lnc)RNA np_5318 has been proved to be involved in renal injury, while its functionality in renal ischemia-reperfusion (I/R) injury is unknown. Therefore, the present study aimed to investigate the role of lncRNA np_5318 in the development of renal I/R injury. Renal I/R injury model a...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
D.A. Spandidos
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7086211/ https://www.ncbi.nlm.nih.gov/pubmed/32256767 http://dx.doi.org/10.3892/etm.2020.8534 |
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author | Lu, Jing Miao, Jiangang Sun, Jianhua |
author_facet | Lu, Jing Miao, Jiangang Sun, Jianhua |
author_sort | Lu, Jing |
collection | PubMed |
description | Long noncoding (Lnc)RNA np_5318 has been proved to be involved in renal injury, while its functionality in renal ischemia-reperfusion (I/R) injury is unknown. Therefore, the present study aimed to investigate the role of lncRNA np_5318 in the development of renal I/R injury. Renal I/R injury model and I/R cell model were established in vitro. The expression of np_5318 in I/R cell was inhibited by small interfering (si)-np_5318 and increased by pc-np_5318. Renal function was detected and evaluated by automatic biochemical tests. Immunohistochemical staining was performed to detect the expression cluster of differentiation (CD)31, transforming growth factor (TGF)-β1 and (mothers against decapentaplegic homolog 3) Smad3 in renal tissue. The interaction between np_5318 and Smad3 was verified by chromatin immunoprecipitation (ChIP). Western blotting was performed to detect the expression levels of TGF-β1, Smad3 and phosphorylated (p)-Smad3 in renal tissue and renal cells. Expression of np_5318 in renal tissue and renal cells was detected by reverse transcription-quantitative PCR. Relative cell viability was confirmed by MTT assay. Renal function was impaired and pathological changes in renal tissue were observed in the renal I/R injury group, indicating the renal I/R injury model was successfully established. Compared with the sham group, the expression level of np_5318 significantly increased in the renal I/R injury group. ChIP data confirmed the interaction between np_5318 and Smad3. The expression of TGF-β1, Smad3 and p-Smad3 in renal tissue was also significantly increased in the renal I/R injury group. Furthermore, the I/R cell model in vitro was successfully constructed and np_5318 in I/R group was significantly increased compared with the control group. Cell growth was significantly suppressed in the I/R group compared with the control group. Additionally, transfection with pc-np_5318 significantly inhibited cell growth of I/R cells at 48 and 72 h. While inhibition of np_5318 by si-np_5318 significantly increased the cell growth of I/R cells at 48 and 72 h. Moreover, the level of TGF-β1, p-Smad3 and Smad3 was significantly increased in the I/R group compared with the control group, and transfection with pc-np_5318 significantly increased the level of TGF-β1, p-Smad3 and Smad3. While inhibition of np_5318 by si-np_5318 significantly suppressed the level of TGF-β1, p-Smad3 and Smad3. LncRNA np_5318 may participate in the development of renal I/R injury through TGF-β/Smad signaling pathway. |
format | Online Article Text |
id | pubmed-7086211 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | D.A. Spandidos |
record_format | MEDLINE/PubMed |
spelling | pubmed-70862112020-04-02 LncRNA np_5318 promotes renal ischemia-reperfusion injury through the TGF-β/Smad signaling pathway Lu, Jing Miao, Jiangang Sun, Jianhua Exp Ther Med Articles Long noncoding (Lnc)RNA np_5318 has been proved to be involved in renal injury, while its functionality in renal ischemia-reperfusion (I/R) injury is unknown. Therefore, the present study aimed to investigate the role of lncRNA np_5318 in the development of renal I/R injury. Renal I/R injury model and I/R cell model were established in vitro. The expression of np_5318 in I/R cell was inhibited by small interfering (si)-np_5318 and increased by pc-np_5318. Renal function was detected and evaluated by automatic biochemical tests. Immunohistochemical staining was performed to detect the expression cluster of differentiation (CD)31, transforming growth factor (TGF)-β1 and (mothers against decapentaplegic homolog 3) Smad3 in renal tissue. The interaction between np_5318 and Smad3 was verified by chromatin immunoprecipitation (ChIP). Western blotting was performed to detect the expression levels of TGF-β1, Smad3 and phosphorylated (p)-Smad3 in renal tissue and renal cells. Expression of np_5318 in renal tissue and renal cells was detected by reverse transcription-quantitative PCR. Relative cell viability was confirmed by MTT assay. Renal function was impaired and pathological changes in renal tissue were observed in the renal I/R injury group, indicating the renal I/R injury model was successfully established. Compared with the sham group, the expression level of np_5318 significantly increased in the renal I/R injury group. ChIP data confirmed the interaction between np_5318 and Smad3. The expression of TGF-β1, Smad3 and p-Smad3 in renal tissue was also significantly increased in the renal I/R injury group. Furthermore, the I/R cell model in vitro was successfully constructed and np_5318 in I/R group was significantly increased compared with the control group. Cell growth was significantly suppressed in the I/R group compared with the control group. Additionally, transfection with pc-np_5318 significantly inhibited cell growth of I/R cells at 48 and 72 h. While inhibition of np_5318 by si-np_5318 significantly increased the cell growth of I/R cells at 48 and 72 h. Moreover, the level of TGF-β1, p-Smad3 and Smad3 was significantly increased in the I/R group compared with the control group, and transfection with pc-np_5318 significantly increased the level of TGF-β1, p-Smad3 and Smad3. While inhibition of np_5318 by si-np_5318 significantly suppressed the level of TGF-β1, p-Smad3 and Smad3. LncRNA np_5318 may participate in the development of renal I/R injury through TGF-β/Smad signaling pathway. D.A. Spandidos 2020-04 2020-02-18 /pmc/articles/PMC7086211/ /pubmed/32256767 http://dx.doi.org/10.3892/etm.2020.8534 Text en Copyright: © Lu et al. This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made. |
spellingShingle | Articles Lu, Jing Miao, Jiangang Sun, Jianhua LncRNA np_5318 promotes renal ischemia-reperfusion injury through the TGF-β/Smad signaling pathway |
title | LncRNA np_5318 promotes renal ischemia-reperfusion injury through the TGF-β/Smad signaling pathway |
title_full | LncRNA np_5318 promotes renal ischemia-reperfusion injury through the TGF-β/Smad signaling pathway |
title_fullStr | LncRNA np_5318 promotes renal ischemia-reperfusion injury through the TGF-β/Smad signaling pathway |
title_full_unstemmed | LncRNA np_5318 promotes renal ischemia-reperfusion injury through the TGF-β/Smad signaling pathway |
title_short | LncRNA np_5318 promotes renal ischemia-reperfusion injury through the TGF-β/Smad signaling pathway |
title_sort | lncrna np_5318 promotes renal ischemia-reperfusion injury through the tgf-β/smad signaling pathway |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7086211/ https://www.ncbi.nlm.nih.gov/pubmed/32256767 http://dx.doi.org/10.3892/etm.2020.8534 |
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