棕榈酸诱导斑马鱼脂毒性损伤骨形成抑制模型的建立
Establishment of a lipotoxic inhibition model of bone formation in zebrafish induced by palmitic acid
王晓仪 1李苗 2王琳霞 2喻斌 2华永庆3
作者信息
- 1. 江苏省中药资源产业化过程协同创新中心,南京 210023;南京中医药大学药学院,南京 210023
- 2. 南京中医药大学药学院,南京 210023
- 3. 江苏省中药资源产业化过程协同创新中心,南京 210023;南京中医药大学药学院,南京 210023;江苏省中药药效与安全性评价重点实验室,南京 210023
- 折叠
摘要
目的 建立棕榈酸诱导的斑马鱼脂毒性损伤骨形成抑制模型.方法 将AB品系斑马鱼胚胎分为对照组、棕榈酸组(PA组)和辛伐他汀组(SIM组).从 3 dpf(days post fertilization,dpf)开始,PA组、SIM组给予PA造模.从 5 dpf开始,SIM组给予SIM连续给药4 d.9 dpf通过钙黄绿素染色法、尼罗红染色法、甘油三酯及总胆固醇含量测定、q-PCR判断模型是否成功建立.结果 PA显著减少斑马鱼椎骨骨节数目、促进脂质堆积、增加甘油三酯及总胆固醇含量、促进成脂相关基因PPARγ、c/EBPα的表达并抑制成骨相关基因ALP、RUNX2 的表达.SIM可以改善PA对斑马鱼的骨形成抑制效应.结论 采用PA给药的方法可成功造成与骨质疏松症(osteoporosis,OP)病理过程相似的脂毒性损伤骨形成抑制模型,该方法简便、灵敏、可控,可用于OP及相关疾病的药物筛选.
Abstract
Objective To establish a palmitic acid(PA)-induced model of lipotoxic bone formation inhibition in zebrafish.Methods AB strain zebrafish embryos were divided randomly into a blank control group,PA group,and simvastatin(SIM)group.Embryos in the PA and SIM groups received PA from 3 days post-fertilization(dpf),and embryos in the SIM group received SIM continuously for 4 days from 5 dpf.Establishment of the model was confirmed at 9 dpf by calcein staining,Nile red staining,triglyceride and total cholesterol content determination,and q-PCR.Results PA significantly decreased the number of vertebrae,promoted lipid accumulation,increased triglyceride and total cholesterol contents,promoted the expression of lipid-related genes PPARγ、c/EBPα,and inhibited the expression of osteogenic genes ALP and RUNX2.SIM improved the inhibitory effect of PA on bone formation in zebrafish.Conclusions PA can successfully create a lipotoxic model of bone-formation inhibition,similar to the pathological process of osteoporosis,using a simple,sensitive,and controllable method.This model can then be used for drug screening for osteoporosis and related diseases.
关键词
骨质疏松模型/脂毒性/斑马鱼/棕榈酸/辛伐他汀Key words
osteoporosis model/lipotoxicity/zebrafish/palmitic acid/simvastatin引用本文复制引用
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)
出版年
2024
国家科技期刊平台
NSTL
万方数据