中国实验动物学报2024，Vol.32Issue(5)：547-556,10.DOI:10.3969/j.issn.1005-4847.2024.05.001
斑马鱼快速骨质疏松模型方法的建立与优化
Establishment and optimization of rapid model of osteoporosis in zebrafish
摘要
Abstract
Objective To establish a fast,stable,and sensitive zebrafish model of osteoporosis(OP)using different method.Methods OP models were induced by iron overload or prednisolone(Pred),and bone formation and mortality were observed.The groups were divided into:Control group,model group(include FAC group and Pred group),and positive control group(AC group).Ammonium ferric citrate was used as the model drug in the iron-overload induction method.For the Pred induction models,the modeling time for the Pred-3 days post-fertilization(dpf)method was 3～9 dpf,the modeling time for the Pred-5 dpf method was 5～10 dpf,and Pred was administered from 3 dpf and removed from 7～9 dpf for the Pred withdrawal method.To compare the anti-osteoporosis(OP)effects of commonly used drugs such as Alfacalcidol(AC),Calcitriol(CA),and Alendronate(AL),it's important to select a stable and sensitive positive control drug and to further optimize different staining methods and conditions.Results There was no significant effect of ammonium ferric citrate 500 μg/mL on bone formation.Bone formation and the length of the first vertebra were significantly decreased in the Pred group induced by Pred-3 dpf compared with those in the control group(P＜0.01,P＜0.05),but zebrafish mortality was higher.There was no significant difference between the Pred-5 dpf method,but bone formation was significantly reduced in the Pred withdrawal group(P＜0.01),with no mortality.Alfacalcidol,calcitriol,and alendronate all had anti-OP effects,with CA having the most sensitive and stable anti-OP effect.Alizarin red staining showed that the optimal dye parameters were 0.02%concentration for dyeing 2 h,with washing in 0.5%KOH and glycerol under the conditions of a 3∶1 ratio for 3 h followed by a 1∶1 ratio for 14 h.The result of staining showed that calcein was more sensitive for staining bone nodes and ARS staining was more sensitive for staining the first vertebra.Conclusions The Pred withdrawal method can be used to establish a rapid,stable,and sensitive OP model in zebrafish as a reliable model for studying OP.关键词
斑马鱼/骨质疏松/泼尼松龙/茜素红/骨化三醇Key words
zebrafish/osteoporosis/prednisolone/alizarin red/calcitriol分类
生物科学引用本文复制引用
毛洪运,刘雨彤,赵心月,姜德利,王晓仪,赵珂璇,华永庆,许惠琴..斑马鱼快速骨质疏松模型方法的建立与优化[J].中国实验动物学报,2024,32(5):547-556,10.基金项目
国家自然科学基金(81473390),江苏省高等学校自然科学研究重大项目(20KJA360001),贵州省科技计划项目([2020]4Y090),贵阳市科技重大项目([2020]-18-4).Funded by the National Natural Science Foundation of China(81473390),the Natural Science Foundation of the Jiangsu Higher Education Institutions of China(20KJA360001),the Science and Technology Program of Guizhou Province([2020]4Y090),the Guiyang Science and Technology Plan Project([2020]-18-4). （81473390）
