摘要
目的探讨糖皮质激素刺激下骨重塑相关细胞外泌体miRNA的变化。方法体外培养小鼠成骨细胞、破骨细胞,提取上清液中外泌体进行透射电镜观察、粒径分析、CD9\CD81表面标记物检测;利用不同浓度(1μg/ml,10μg/ml,100μg/ml)地塞米松刺激48 h,观察两种细胞增殖情况;收集外泌体检测骨重塑相关miR-133、miR-214、miR-4697和miR-487的表达。结果不同浓度地塞米松均能够抑制两种细胞增殖,其中100μg/ml组与对照组相比抑制作用显著(P<0.05)。当地塞米松浓度为100μg/ml时,成骨细胞外泌体miR-133含量降低为对照组的45%,miR-214降为对照组的12%,miR-4697降为对照组的56%(P均<0.05)。当地塞米松浓度为1μg/ml时,破骨细胞外泌体内miR-133和miR-487上升为对照组的12倍,miR-214升高为对照组1.67倍,miR-487升高2.12倍(P均<0.05)。结论随着糖皮质激素浓度增加,成骨细胞外泌体内miR-133、miR-214、miR-4697含量依次递减,而miR-487含量随着糖皮质激素浓度先升高后降低。小剂量糖皮质激素刺激破骨细胞后,其外泌体内4种miRNA含量均增加,而高浓度糖皮质激素刺激后miRNA含量降低。
Objective To investigate the changes of mi RNAs in exosomes derived from bone remodeling-related cells stimulated by glucocorticoid. Methods Osteoblasts and osteoclasts were cultured in vitro. Exosomes from the supernatant were collected for transmission electron microscopy, particle size analysis and CD9/CD81 surface marker detection. The proliferation of two kinds of cells was observed using CCK-8 assay after stimulating by different concentrations of dexamethasone(1μg/ml, 10μg/ml,100μg/ml) for 48 hours. Exosomes were extracted to detect the expression of mi R-133, mi R-214 and mi R-4697 and mi R-487.Results The proliferation of both cells could be inhibited by different concentrations of dexamethasone, of which 100μg/ml of dexamethasone had the strongest inhibitory effect. When the concentration of dexamethasone was 100μg/ml, the level of mi R-133 in osteoblast exosomes reduced to 45% compared with the control group, meanwhile mi R-214 and mi R-4697 reduced to 12% and 56%, respectively(P〈0.05). Compared with the control group, when the concentration of dexamethasone was 1μg/ml, the expression of both mi R-133 and mi R-487 increased 12-fold, and the expression of mi R-214 increased 1.67-fold and mi R-487 increased 2.12-fold. Conclusion With the increase of glucocorticoid concentration, the content of mi R-133, mi R-214 and mi R-4697 decrease gradually in osteoblast exosomes, while the content of mi R-487 increase first and then decrease. After stimulating osteoclasts with low dose glucocorticoid, the content of four mi RNAs in exosomes increases, while it decreases in high dose glucocorticoids group.
作者
李毅
尹鹏滨
邓元
孟钰童
吕厚辰
张里程
唐佩福
LI Yi;YIN Pengbin;DENG Yuan;MENG Yutong;LYU Houchen;ZHANG Licheng;TANG Peifu(Chinese PLA General Hospital, Beijing 100853, China)
出处
《解放军医学院学报》
CAS
2018年第4期344-348,共5页
Academic Journal of Chinese PLA Medical School