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

作　　者：王晓仪 李苗 王琳霞 喻斌[2] 华永庆 WANG Xiaoyi;LI Miao;WANG Linxia;YU Bin;HUA Yongqing(Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization,Nanjing 210023,China;College of Pharmacy,Nanjing University of Chinese Medicine,Nanjing 210023,China;Jiangsu Key Laboratory for Pharmacology and Safety Evaluation of Chinese Materia Medica,Nanjing 210023,China)

机构地区：[1]江苏省中药资源产业化过程协同创新中心,南京210023 [2]南京中医药大学药学院,南京210023 [3]江苏省中药药效与安全性评价重点实验室,南京210023

出　　处：《中国实验动物学报》2024年第4期461-467,共7页Acta Laboratorium Animalis Scientia Sinica

基　　金：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)。

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。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 

分 类 号：Q95-33[生物学—动物学]