MiR-761 promotes aortic valve calcification by inhibiting GABARAP expression
Objective To investigate the effect and potential mechanism of miR-761 on calcified aortic valve disease (CAVD). Methods Normal (n=6) and calcified (n=8) valve tissues were collected from Department of Cardiothoracic Surgery of the First Affiliated Hospital of Chongqing Medical University between Ja...
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| Format: | Article |
| Language: | zho |
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Editorial Office of Journal of Army Medical University
2023-08-01
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| Series: | 陆军军医大学学报 |
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| Online Access: | http://aammt.tmmu.edu.cn/html/202303056.htm |
| _version_ | 1797731219204472832 |
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| author | XIANG Yi LI Xiaorong LUO Jiangqiao HE Wenhuan |
| author_facet | XIANG Yi LI Xiaorong LUO Jiangqiao HE Wenhuan |
| author_sort | XIANG Yi |
| collection | DOAJ |
| description | Objective To investigate the effect and potential mechanism of miR-761 on calcified aortic valve disease (CAVD). Methods Normal (n=6) and calcified (n=8) valve tissues were collected from Department of Cardiothoracic Surgery of the First Affiliated Hospital of Chongqing Medical University between January 2022 and January 2023. The changes of autophagy, osteogenesis, GABARAP protein or miR-761 expression in calcified valve tissues were detected by Western blotting, immunohistochemical staining and RT-qPCR. Double luciferase assay verified miR-761 binding to GABARAP 3′-UTR region. The phenotypes of porcine aortic valve interstitial cells (pVICs) were identified by immunofluorescence assay. pVICs were transfected with miR-761 mimic, inhibitor and corresponding control to construct pVICs cell lines with over-expression or knockdown of miR-761. After adding GFP-LC3 adenovirus, the number of autophagosomes was observed. Western blotting, alkaline phosphatase staining and alizarin red S staining were used to detect the osteogenic differentiation of pVICs before and after osteogenic medium(OM) treatment. Results Compared with normal tissues, the prdein levels of Beclin 1, ULK1, α-SMA, RUNX2 and GABARAP were up-regulated in calcified valve tissue tissues, while the levels of p62 and miR-761 were down-regulated (P < 0.05). Dual luciferase assay verified the binding of miR-761 to GABARAP 3′-UTR region (P < 0.05). The isolated pVICs expressed interstitial cell markers (α-SMA, Vimentin). Confocal microscopy showed that overexpression of miR-761 reduced autophagosome production in pVICs. The protein results, alkaline phosphatase staining and alizarin red S staining showed that knockdown of miR-761 significantly increased the expression of GABARAP in pVICs, significantly decreased the expressions of OPN, α-SMA, Collagen Ⅰ and RUNX2 (P < 0.05), and inhibited the calcium salt deposition in the cells induced by OM. Conclusion MiR-761 may inhibit autophagy and promote cardiac valve calcification by inhibiting GABARAP expression.
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| first_indexed | 2024-03-12T11:55:38Z |
| format | Article |
| id | doaj.art-53754139fc5c47ecaa92c159cbc7b469 |
| institution | Directory Open Access Journal |
| issn | 2097-0927 |
| language | zho |
| last_indexed | 2024-03-12T11:55:38Z |
| publishDate | 2023-08-01 |
| publisher | Editorial Office of Journal of Army Medical University |
| record_format | Article |
| series | 陆军军医大学学报 |
| spelling | doaj.art-53754139fc5c47ecaa92c159cbc7b4692023-08-31T01:06:38ZzhoEditorial Office of Journal of Army Medical University陆军军医大学学报2097-09272023-08-0145161702171110.16016/j.2097-0927.202303056MiR-761 promotes aortic valve calcification by inhibiting GABARAP expressionXIANG Yi0LI Xiaorong1LUO Jiangqiao2HE Wenhuan3College of Laboratory Medicine, Key Laboratory of Laboratory Medicine Diagnostics of Ministry of Education, Chongqing Medical University, Chongqing 400016College of Laboratory Medicine, Key Laboratory of Laboratory Medicine Diagnostics of Ministry of Education, Chongqing Medical University, Chongqing 400016College of Laboratory Medicine, Key Laboratory of Laboratory Medicine Diagnostics of Ministry of Education, Chongqing Medical University, Chongqing 400016College of Laboratory Medicine, Key Laboratory of Laboratory Medicine Diagnostics of Ministry of Education, Chongqing Medical University, Chongqing 400016Objective To investigate the effect and potential mechanism of miR-761 on calcified aortic valve disease (CAVD). Methods Normal (n=6) and calcified (n=8) valve tissues were collected from Department of Cardiothoracic Surgery of the First Affiliated Hospital of Chongqing Medical University between January 2022 and January 2023. The changes of autophagy, osteogenesis, GABARAP protein or miR-761 expression in calcified valve tissues were detected by Western blotting, immunohistochemical staining and RT-qPCR. Double luciferase assay verified miR-761 binding to GABARAP 3′-UTR region. The phenotypes of porcine aortic valve interstitial cells (pVICs) were identified by immunofluorescence assay. pVICs were transfected with miR-761 mimic, inhibitor and corresponding control to construct pVICs cell lines with over-expression or knockdown of miR-761. After adding GFP-LC3 adenovirus, the number of autophagosomes was observed. Western blotting, alkaline phosphatase staining and alizarin red S staining were used to detect the osteogenic differentiation of pVICs before and after osteogenic medium(OM) treatment. Results Compared with normal tissues, the prdein levels of Beclin 1, ULK1, α-SMA, RUNX2 and GABARAP were up-regulated in calcified valve tissue tissues, while the levels of p62 and miR-761 were down-regulated (P < 0.05). Dual luciferase assay verified the binding of miR-761 to GABARAP 3′-UTR region (P < 0.05). The isolated pVICs expressed interstitial cell markers (α-SMA, Vimentin). Confocal microscopy showed that overexpression of miR-761 reduced autophagosome production in pVICs. The protein results, alkaline phosphatase staining and alizarin red S staining showed that knockdown of miR-761 significantly increased the expression of GABARAP in pVICs, significantly decreased the expressions of OPN, α-SMA, Collagen Ⅰ and RUNX2 (P < 0.05), and inhibited the calcium salt deposition in the cells induced by OM. Conclusion MiR-761 may inhibit autophagy and promote cardiac valve calcification by inhibiting GABARAP expression. http://aammt.tmmu.edu.cn/html/202303056.htmmir-761gamma-aminobutyric acid receptor-associated proteinautophagycalcified aortic valve disease |
| spellingShingle | XIANG Yi LI Xiaorong LUO Jiangqiao HE Wenhuan MiR-761 promotes aortic valve calcification by inhibiting GABARAP expression 陆军军医大学学报 mir-761 gamma-aminobutyric acid receptor-associated protein autophagy calcified aortic valve disease |
| title | MiR-761 promotes aortic valve calcification by inhibiting GABARAP expression |
| title_full | MiR-761 promotes aortic valve calcification by inhibiting GABARAP expression |
| title_fullStr | MiR-761 promotes aortic valve calcification by inhibiting GABARAP expression |
| title_full_unstemmed | MiR-761 promotes aortic valve calcification by inhibiting GABARAP expression |
| title_short | MiR-761 promotes aortic valve calcification by inhibiting GABARAP expression |
| title_sort | mir 761 promotes aortic valve calcification by inhibiting gabarap expression |
| topic | mir-761 gamma-aminobutyric acid receptor-associated protein autophagy calcified aortic valve disease |
| url | http://aammt.tmmu.edu.cn/html/202303056.htm |
| work_keys_str_mv | AT xiangyi mir761promotesaorticvalvecalcificationbyinhibitinggabarapexpression AT lixiaorong mir761promotesaorticvalvecalcificationbyinhibitinggabarapexpression AT luojiangqiao mir761promotesaorticvalvecalcificationbyinhibitinggabarapexpression AT hewenhuan mir761promotesaorticvalvecalcificationbyinhibitinggabarapexpression |