Experimental study of Rac1 regulating osteoclast function in hypoxic environment
[Objective:] To investigate the effect of hypoxia-inducible factor-1α (HIF-1α) on the changes of osteoclast bone resorption capacity by regulating Rac1 on cytoskeleton under hypoxic environment. [Methods:] The hypoxia culture system was established by stimulating mouse mononuclear cell line RAW264.7...
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Editorial Office of Journal of Oral and Maxillofacial Surgery
2024-02-01
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| Series: | Kouqiang hemian waike zazhi |
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| Online Access: | https://journal06.magtech.org.cn/Jweb_joms/EN/10.12439/kqhm.1005-4979.2024.01.002 |
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| _version_ | 1849225127816855552 |
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| author | GU Jiahong TIAN Yuanye KANG Feiwu |
| author_facet | GU Jiahong TIAN Yuanye KANG Feiwu |
| author_sort | GU Jiahong |
| collection | DOAJ |
| description | [Objective:] To investigate the effect of hypoxia-inducible factor-1α (HIF-1α) on the changes of osteoclast bone resorption capacity by regulating Rac1 on cytoskeleton under hypoxic environment. [Methods:] The hypoxia culture system was established by stimulating mouse mononuclear cell line RAW264.7 with cobalt dichloride (CoCl2) in vitro. The differentiation of RAW264.7 cells was induced by sRANKL conditioned culture medium and stimulated by CoCl2 simulated hypoxic environment, and TRAP staining was performed after 7 days; real-time quantitative polymerase chain reaction (RT-qPCR), cellular immunofluorescence assay and Western blotting were used to detect the expression of HIF-1α KO Rac1, osteoclast-related surface markers and closure zones in RAW264.7 under hypoxia. Simultaneously, HIF-1α conditional knockout cell line RAW264.7 and the effect of the inhibitor of Rac1 on the expression of osteoclast-related genes and closure zones in RAW264.7 under the same conditions were detected. [Results:] Hypoxia could increase the volume of RAW264.7 induced osteoclasts and enhance bone resorption capacity (P<0.01). The level of Rac1 mRNA expressed by osteoclasts in hypoxia group was increased, and the expression of closure zones were significant (P<0.001). The volume of osteoclasts induced by HIF-1α KO RAW264.7 cells decreased, the expression of HIF-1α was significantly decreased, and Rac1 was also decreased (P<0.001); meanwhile, the volume of osteoclasts decreased in the inhibitor group, the expression of HIF-1α remained unchanged, and the closure zones formed abnormally (P<0.01). [Conclusion:] Under hypoxia, mouse mononuclear cell line RAW264.7 can regulate Rac1 through HIF-1α protein to affect the cytoskeleton of osteoclasts, and then change the bone resorption capacity of osteoclasts. |
| format | Article |
| id | doaj-art-c1e1093ca6794e57bf657403a4d2a482 |
| institution | Kabale University |
| issn | 1005-4979 |
| language | zho |
| publishDate | 2024-02-01 |
| publisher | Editorial Office of Journal of Oral and Maxillofacial Surgery |
| record_format | Article |
| series | Kouqiang hemian waike zazhi |
| spelling | doaj-art-c1e1093ca6794e57bf657403a4d2a4822025-08-25T06:10:50ZzhoEditorial Office of Journal of Oral and Maxillofacial SurgeryKouqiang hemian waike zazhi1005-49792024-02-01341142110.12439/kqhm.1005-4979.2024.01.002Experimental study of Rac1 regulating osteoclast function in hypoxic environmentGU Jiahong0TIAN Yuanye1KANG Feiwu2Department of Oral and Maxillofacial Surgery, Stomatological Hospital and Dental School of Tongji University, Shanghai Engineering Research Center of Tooth Restoration and Regeneration, Shanghai 200072Department of Oral and Maxillofacial Surgery, Hospital of Stomatology, Jilin University, Changchun 130021, ChinaDepartment of Oral and Maxillofacial Surgery, Stomatological Hospital and Dental School of Tongji University, Shanghai Engineering Research Center of Tooth Restoration and Regeneration, Shanghai 200072[Objective:] To investigate the effect of hypoxia-inducible factor-1α (HIF-1α) on the changes of osteoclast bone resorption capacity by regulating Rac1 on cytoskeleton under hypoxic environment. [Methods:] The hypoxia culture system was established by stimulating mouse mononuclear cell line RAW264.7 with cobalt dichloride (CoCl2) in vitro. The differentiation of RAW264.7 cells was induced by sRANKL conditioned culture medium and stimulated by CoCl2 simulated hypoxic environment, and TRAP staining was performed after 7 days; real-time quantitative polymerase chain reaction (RT-qPCR), cellular immunofluorescence assay and Western blotting were used to detect the expression of HIF-1α KO Rac1, osteoclast-related surface markers and closure zones in RAW264.7 under hypoxia. Simultaneously, HIF-1α conditional knockout cell line RAW264.7 and the effect of the inhibitor of Rac1 on the expression of osteoclast-related genes and closure zones in RAW264.7 under the same conditions were detected. [Results:] Hypoxia could increase the volume of RAW264.7 induced osteoclasts and enhance bone resorption capacity (P<0.01). The level of Rac1 mRNA expressed by osteoclasts in hypoxia group was increased, and the expression of closure zones were significant (P<0.001). The volume of osteoclasts induced by HIF-1α KO RAW264.7 cells decreased, the expression of HIF-1α was significantly decreased, and Rac1 was also decreased (P<0.001); meanwhile, the volume of osteoclasts decreased in the inhibitor group, the expression of HIF-1α remained unchanged, and the closure zones formed abnormally (P<0.01). [Conclusion:] Under hypoxia, mouse mononuclear cell line RAW264.7 can regulate Rac1 through HIF-1α protein to affect the cytoskeleton of osteoclasts, and then change the bone resorption capacity of osteoclasts.https://journal06.magtech.org.cn/Jweb_joms/EN/10.12439/kqhm.1005-4979.2024.01.002cytoskeletonhypoxia-inducible factor-1αraw264.7 cellsrac1 |
| spellingShingle | GU Jiahong TIAN Yuanye KANG Feiwu Experimental study of Rac1 regulating osteoclast function in hypoxic environment Kouqiang hemian waike zazhi cytoskeleton hypoxia-inducible factor-1α raw264.7 cells rac1 |
| title | Experimental study of Rac1 regulating osteoclast function in hypoxic environment |
| title_full | Experimental study of Rac1 regulating osteoclast function in hypoxic environment |
| title_fullStr | Experimental study of Rac1 regulating osteoclast function in hypoxic environment |
| title_full_unstemmed | Experimental study of Rac1 regulating osteoclast function in hypoxic environment |
| title_short | Experimental study of Rac1 regulating osteoclast function in hypoxic environment |
| title_sort | experimental study of rac1 regulating osteoclast function in hypoxic environment |
| topic | cytoskeleton hypoxia-inducible factor-1α raw264.7 cells rac1 |
| url | https://journal06.magtech.org.cn/Jweb_joms/EN/10.12439/kqhm.1005-4979.2024.01.002 |
| work_keys_str_mv | AT gujiahong experimentalstudyofrac1regulatingosteoclastfunctioninhypoxicenvironment AT tianyuanye experimentalstudyofrac1regulatingosteoclastfunctioninhypoxicenvironment AT kangfeiwu experimentalstudyofrac1regulatingosteoclastfunctioninhypoxicenvironment |