中国实验动物学报2025，Vol.33Issue(9)：1259-1269,11.DOI:10.3969/j.issn.1005-4847.2025.09.002
地塞米松诱导糖皮质激素性骨质疏松与高血压双重表型斑马鱼模型的构建与评价
Construction and evaluation of a zebrafish model of dexamethasone-induced osteoporosis combined with hypertension
摘要
Abstract
Objective To establish a dexamethasone(Dex)-induced zebrafish model of glucocorticoid-induced osteoporosis(GIOP)combined with glucocorticoid-induced hypertension(GIHT),and to validate the model by the systematic evaluation of both the phenotypic manifestations and molecular mechanisms.Methods Zebrafish larvae at 3 or 4 d post-fertilization(dpf)were divided randomly into a control group(0.1％dimethyl sulfoxide)and a model group(10 μmol/L Dex).Osteogenic parameters and vessel diameter were assessed at 0,48,and 96 h post-administration(n=10).Bone mineralization and density were determined by the total area and sum brightness after Alizarin red staining.Vessel diameter was measured by detecting blood flow in the dorsal aorta.After confirming the optimal administration time,expression levels of bone-formation-related proteins(protein kinase B(Akt),glycogen synthase kinase(GSK)-3β,β-catenin)and angiogenesis-related proteins(AMP-activated protein kinase(AMPK),nuclear factor(NF)-κB)were detected by Western Blot to verify the molecular effectiveness of the model.Results Exposure to Dex for 96 h reduced bone mineralization and density in zebrafish larvae compared with the control group,and statistical analysis identified 4 dpf zebrafish and Dex administration for 96 h as the optimal modeling times for the GIOP model.Blood vessel diameter was significantly decreased in the model group compared with the control group(P＜0.05),and the difference became more pronounced with longer administration time and was particularly evident at 4 dpf and treatment for 96 h.Western Blot analysis showed that Dex significantly decreased protein expression levels of Akt,β-catenin,and NF-κB(P＜0.05)and significantly increased the expression of GSK-3β and AMPK(P＜0.05),suggesting that Dex effectively inhibited bone formation and angiogenesis after 96 hours treatment in 4 dpf zebrafish.Conclusions Treatment of 4 dpf zebrafish larvae with 10 μmol/L Dex rapidly established a reliable zebrafish model of GIOP combined with GIHT,providing an ideal animal model for further studies of the common mechanisms of the two diseases and for screening new drugs.关键词
斑马鱼/地塞米松/糖皮质激素性骨质疏松症/糖皮质激素性高血压/双重表型模型构建Key words
zebrafish/dexamethasone/glucocorticoid-induced osteoporosis/glucocorticoid-induced hypertension/dual-phenotype model construction分类
生物科学引用本文复制引用
谢安娜,曹金龙,戴薇薇..地塞米松诱导糖皮质激素性骨质疏松与高血压双重表型斑马鱼模型的构建与评价[J].中国实验动物学报,2025,33(9):1259-1269,11.基金项目
国家自然科学基金(81873318,82374474),上海中医药大学医养结合科创项目(YYKC-2021-01-010),上海中医药大学附属龙华医院教育教学改革项目(2023lhjx043).Funded by the National Natural Science Foundation of China(81873318,82374474),the Project Combining Medical Health and Nursing of Shanghai University of Traditional Chinese Medieine(YYKC-2021-01-010),the Education and Teaching Reform Project of Longhua Hospital Shanghai University of Traditional Chinese Medicine(2023lhjx043). （81873318,82374474）
