He S, Wen N, Chen X, Wang Y, Zhang T, Mu Y
Cheng Fei Hospital, Chengdu 610073, China.
The First People's Hospital of Yibin, Yibin 644000, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2024 Sep 20;44(9):1752-1759. doi: 10.12122/j.issn.1673-4254.2024.09.15.
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.
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.
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 -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.
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.
探讨枸杞糖肽(LbGP)对受辐射人牙龈成纤维细胞(HGFs)来源的外泌体诱导的成骨抑制的保护作用。
对培养的HGFs进行或不进行LbGP预处理后,给予8 Gy X射线辐射,采用RT-qPCR、蛋白质免疫印迹法和β-半乳糖苷酶染色检测细胞凋亡、衰老及α-SMA水平的变化。提取处理后细胞分泌的外泌体,经电子显微镜、粒径分析和蛋白质免疫印迹法鉴定后,将外泌体加入原代培养的骨髓间充质干细胞(BMSCs)中,通过抗酒石酸酸性磷酸酶染色、茜素红染色、碱性磷酸酶染色、RT-qPCR和蛋白质免疫印迹法检测细胞培养中的破骨细胞活性和成骨情况。
在培养的HGFs中,X射线辐射显著增加了衰老细胞的百分比,LbGP处理可明显降低该百分比。X射线辐射显著降低了HGFs中Bcl-2/Bax比值并增加了α-SMA表达,而LbGP预处理可显著抑制这些变化。在大鼠BMSCs中,与受辐射HGFs来源的外泌体共孵育导致破骨细胞显著增加,钙结节减少,细胞碱性磷酸酶表达降低;在用LbGP预处理的HGFs来源的外泌体处理的细胞中观察到相反的变化,与受辐射HGFs来源的外泌体相比,其还显著增加了成骨基因(BMP2、ALP和RUNX2)和蛋白质(ALP和RUNX2)的细胞表达。
LbGP可通过作用于受辐射HGFs分泌的外泌体有效抑制破骨细胞活性并促进成骨,提示其在治疗颌骨放射性骨坏死方面的潜在价值。