Circ‐NCX1 inhibits LPS‐induced chondrocyte apoptosis by regulating the miR‐133a/SIRT1 axis
Abstract Osteoarthritis (OA) is a degenerative joint disease, which is characterized by the degeneration of articular cartilage, thickening of subchondral bone, and inflammation of the synovial membrane. In this study, we aimed to investigate the effects and underlying mechanisms of circ‐NCX1 in lip...
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| Format: | Article |
| Language: | English |
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Wiley
2022-10-01
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| Series: | Kaohsiung Journal of Medical Sciences |
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| Online Access: | https://doi.org/10.1002/kjm2.12564 |
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| author | Kai Liu Xiao‐E Fan Li Zhang Ying Yang Xiao‐Ling Zhou |
| author_facet | Kai Liu Xiao‐E Fan Li Zhang Ying Yang Xiao‐Ling Zhou |
| author_sort | Kai Liu |
| collection | DOAJ |
| description | Abstract Osteoarthritis (OA) is a degenerative joint disease, which is characterized by the degeneration of articular cartilage, thickening of subchondral bone, and inflammation of the synovial membrane. In this study, we aimed to investigate the effects and underlying mechanisms of circ‐NCX1 in lipopolysaccharide (LPS)‐induced injury in SW1353 chondrocytes, an in vitro model of OA. The levels of circ‐NCX1, miR‐133a, and related apoptotic proteins were determined by RT‐qPCR. MTT assay was used to evaluate the cell viability. The apoptosis was determined by flow cytometry, whereas the expression of apoptosis proteins was detected by Western blot. Immunofluorescence was used to detect cleaved caspase‐3 expression in cells. Luciferase reporter assay was used to verify the interaction between circ‐NCX1 and miR‐133a, and between miR‐133a and Silent information regulator 2 homolog 1 (Sirt1). The results showed that the overexpression of circ‐NCX1 significantly upregulated the chondrocyte viability and proliferation, and alleviated apoptosis in LPS‐induced SW1353 cells. Immunofluorescence results showed that the overexpression of circ‐NCX1 significantly reduced expression of LPS‐stimulated cleaved Caspase‐3. The RT‐qPCR results showed that the overexpression of circ‐NCX1 inhibited mRNA levels of cleaved Caspase‐3 and Bax, and promoted mRNA levels of Bcl‐2. Luciferase reporter assay showed that circ‐NCX1 targeted miR‐133a, and miR‐133a directly targeted the Sirt1. In addition, overexpression of circ‐NCX1 inhibited chondrocyte apoptosis and promoted Akt phosphorylation via the miR‐133a/Sirt1 axis in LPS‐induced chondrocytes. In conclusion, circ‐NCX1 may serve as an important regulator of LPS‐induced chondrocyte apoptosis through the miR‐133a/Sirt1 axis, and may be involved in the development of OA. |
| first_indexed | 2024-04-12T00:38:00Z |
| format | Article |
| id | doaj.art-f0ec25922db944088fa56d133532402f |
| institution | Directory Open Access Journal |
| issn | 1607-551X 2410-8650 |
| language | English |
| last_indexed | 2024-04-12T00:38:00Z |
| publishDate | 2022-10-01 |
| publisher | Wiley |
| record_format | Article |
| series | Kaohsiung Journal of Medical Sciences |
| spelling | doaj.art-f0ec25922db944088fa56d133532402f2022-12-22T03:55:06ZengWileyKaohsiung Journal of Medical Sciences1607-551X2410-86502022-10-013810992100010.1002/kjm2.12564Circ‐NCX1 inhibits LPS‐induced chondrocyte apoptosis by regulating the miR‐133a/SIRT1 axisKai Liu0Xiao‐E Fan1Li Zhang2Ying Yang3Xiao‐Ling Zhou4Department of Orthopedics The First Affiliated Hospital of Xi'an Jiaotong University Xi'an City Shaanxi Province ChinaDepartment of Thoracic Surgery First Affiliated Hospital of Xi'an Jiaotong University Xi'an City Shaanxi Province ChinaDepartment of Gastroenterology First Affiliated Hospital of Xi'an Jiaotong University Xi'an City Shaanxi Province ChinaYan'an University Yan'an City Shaanxi Province ChinaDepartment of Orthopedics The First Affiliated Hospital of Xi'an Jiaotong University Xi'an City Shaanxi Province ChinaAbstract Osteoarthritis (OA) is a degenerative joint disease, which is characterized by the degeneration of articular cartilage, thickening of subchondral bone, and inflammation of the synovial membrane. In this study, we aimed to investigate the effects and underlying mechanisms of circ‐NCX1 in lipopolysaccharide (LPS)‐induced injury in SW1353 chondrocytes, an in vitro model of OA. The levels of circ‐NCX1, miR‐133a, and related apoptotic proteins were determined by RT‐qPCR. MTT assay was used to evaluate the cell viability. The apoptosis was determined by flow cytometry, whereas the expression of apoptosis proteins was detected by Western blot. Immunofluorescence was used to detect cleaved caspase‐3 expression in cells. Luciferase reporter assay was used to verify the interaction between circ‐NCX1 and miR‐133a, and between miR‐133a and Silent information regulator 2 homolog 1 (Sirt1). The results showed that the overexpression of circ‐NCX1 significantly upregulated the chondrocyte viability and proliferation, and alleviated apoptosis in LPS‐induced SW1353 cells. Immunofluorescence results showed that the overexpression of circ‐NCX1 significantly reduced expression of LPS‐stimulated cleaved Caspase‐3. The RT‐qPCR results showed that the overexpression of circ‐NCX1 inhibited mRNA levels of cleaved Caspase‐3 and Bax, and promoted mRNA levels of Bcl‐2. Luciferase reporter assay showed that circ‐NCX1 targeted miR‐133a, and miR‐133a directly targeted the Sirt1. In addition, overexpression of circ‐NCX1 inhibited chondrocyte apoptosis and promoted Akt phosphorylation via the miR‐133a/Sirt1 axis in LPS‐induced chondrocytes. In conclusion, circ‐NCX1 may serve as an important regulator of LPS‐induced chondrocyte apoptosis through the miR‐133a/Sirt1 axis, and may be involved in the development of OA.https://doi.org/10.1002/kjm2.12564chondrocyte apoptosiscirc‐NCX1miR‐133aosteoarthritisSirt1 |
| spellingShingle | Kai Liu Xiao‐E Fan Li Zhang Ying Yang Xiao‐Ling Zhou Circ‐NCX1 inhibits LPS‐induced chondrocyte apoptosis by regulating the miR‐133a/SIRT1 axis Kaohsiung Journal of Medical Sciences chondrocyte apoptosis circ‐NCX1 miR‐133a osteoarthritis Sirt1 |
| title | Circ‐NCX1 inhibits LPS‐induced chondrocyte apoptosis by regulating the miR‐133a/SIRT1 axis |
| title_full | Circ‐NCX1 inhibits LPS‐induced chondrocyte apoptosis by regulating the miR‐133a/SIRT1 axis |
| title_fullStr | Circ‐NCX1 inhibits LPS‐induced chondrocyte apoptosis by regulating the miR‐133a/SIRT1 axis |
| title_full_unstemmed | Circ‐NCX1 inhibits LPS‐induced chondrocyte apoptosis by regulating the miR‐133a/SIRT1 axis |
| title_short | Circ‐NCX1 inhibits LPS‐induced chondrocyte apoptosis by regulating the miR‐133a/SIRT1 axis |
| title_sort | circ ncx1 inhibits lps induced chondrocyte apoptosis by regulating the mir 133a sirt1 axis |
| topic | chondrocyte apoptosis circ‐NCX1 miR‐133a osteoarthritis Sirt1 |
| url | https://doi.org/10.1002/kjm2.12564 |
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