[枸杞糖肽可减少由辐射照射下的人牙龈成纤维细胞外泌体引起的骨质流失】。]

S He, N Wen, X Chen, Y Wang, T Zhang, Y Mu
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引用次数: 0

摘要

目的探讨枸杞糖肽(LbGP)对暴露于辐射的人牙龈成纤维细胞(HGFs)外泌体诱导的成骨抑制的保护作用:采用RT-qPCR、Western印迹和β-半乳糖苷酶染色检测细胞凋亡、衰老和α-SMA水平的变化。提取处理细胞分泌的外泌体,经电镜鉴定、粒度分析和 Western 印迹后,将外泌体加入原代培养的骨间充质干细胞(BMSCs)中,通过 Trap 染色、茜素红染色、ALP 染色、RT-qPCR 和 Western 印迹检测细胞培养物中破骨细胞的活性和成骨情况:结果:在培养的 HGFs 中,X 射线辐射显著增加了衰老细胞的比例,而 LbGP 处理则明显降低了这一比例。X射线辐射明显降低了HGFs中Bcl-2/Bax的比例,增加了α -SMA的表达,LbGP预处理明显抑制了这些变化。在大鼠 BMSCs 中,用从 HGFs 中提取的外泌体与辐射照射一起孵育会导致细胞中破骨细胞明显增加、钙结节减少和碱性磷酸酶表达降低;与经过辐照的 HGFs 外泌体相比,经过 LbGP 处理的 HGFs 外泌体也能显著增加成骨基因(BMP2、ALP 和 RUNX2)和蛋白质(ALP 和 RUNX2)在细胞中的表达。结论LbGP可通过作用于辐照HGFs分泌的外泌体,有效抑制破骨细胞的活性并促进成骨,这表明它具有治疗颌骨坏死的潜在价值。
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[Lycium barbarum glycopeptide reduces bone loss caused by exosomes derived from human gingival fibroblasts with radiation exposure].

Objective: To explore the protective effect of Lycium barbanun glycopeptide (LbGP) against osteogenic inhibition induced by exosomes derived from human gingival fibroblasts (HGFs) exposed to radiation.

Methods: Cultured HGFs with or without LbGP pretreatment were exposed to 8 Gy X-ray radiation, and the changes in cell apoptosis, senescence and α-SMA level were detected using RT-qPCR, Western blotting and β-galactosidase staining. The exosomes secreted by the treated cells were extracted, and after identification by electron microscopy, particle size analysis and Western blotting, the exosomes were added into primary cultured bone mesenchymal stem cells (BMSCs), and osteoclast activity and osteogenesis in the cell cultures were detected by Trap staining, Alizarin red staining, ALP staining, RT-qPCR and Western blotting.

Results: In cultured HGFs, X-ray radiation significantly increased the percentage of senescent cells, which was obviously lowered by LbGP treatment. X-ray radiation significantly reduced Bcl-2/Bax ratio and increased α -SMA expression in HGFs, and these changes were significantly suppressed by LbGP pretreatment. In rat BMSCs, incubation with the exosomes derived from HGFs with radiation exposure caused a significant increase of osteoclasts, reduced calcium nodules and lowered alkaline phosphatase expression in the cells; The opposite changes were observed in the cells treated with exosomes from LbGPpretreated HGFs, which also significantly increased the cellular expressions of the osteogenic genes (BMP2, ALP, and RUNX2) and proteins (ALP and RUNX2) as compared with the exosomes from irradiated HGFs.

Conclusion: LbGP can effectively inhibit osteoclast activity and promote osteogenesis by acting on exosomes secreted by irradiated HGFs, suggesting its potential value for treatment of osteoradionecrosis of the jaw.

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来源期刊
CiteScore
1.50
自引率
0.00%
发文量
208
期刊最新文献
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